Steroids 77 (2012) 10–26

Contents lists available at SciVerse ScienceDirect

Steroids
journal homepage: www.elsevier.com/locate/steroids

Review

Oxidative metabolism of dehydroepiandrosterone (DHEA) and biologically

active oxygenated metabolites of DHEA and epiandrosterone (EpiA) – Recent reports
Laïla El Kihel ⇑
Université de Caen Basse-Normandie, U.F.R. des Sciences Pharmaceutiques, Centre d’Etudes et de Recherche sur le Médicament de Normandie, UPRES EA-4258,
FR CNRS INC3M, Boulevard Becquerel, 14032 Caen Cedex, France

a r t i c l e i n f o a b s t r a c t

Article history: Dehydroepiandrosterone (DHEA) is a multifunctional steroid with a broad range of biological effects in
Received 21 April 2011 humans and animals. DHEA can be converted to multiple oxygenated metabolites in the brain and
Received in revised form 14 September peripheral tissues. The mechanisms by which DHEA exerts its effects are not well understood. However,
2011
evidence that the effects of DHEA are mediated by its oxygenated metabolites has accumulated.
Accepted 18 September 2011
Available online 2 October 2011
This paper will review the panel of oxygenated DHEA metabolites (7, 16 and 17-hydroxylated deriva-
tives) including a number of 5a-androstane derivatives, such as epiandrosterone (EpiA) metabolites. The
most important aspects of the oxidative metabolism of DHEA in the liver, intestine and brain are
Keywords:
Oxygenated DHEA metabolites
described. Then, this article reviews the reported biological effects of oxygenated DHEA metabolites from
Epiandrosterone recent findings with a specific focus on cancer, inflammatory and immune processes, osteoporosis, ther-
Oxidative DHEA metabolism mogenesis, adipogenesis, the cardiovascular system, the brain and the estrogen and androgen receptors.
Hydroxy-DHEA Ó 2011 Elsevier Inc. All rights reserved.
Hydroxy-EpiA
DHEA metabolites activities

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
2. Oxidative metabolism of DHEA and epiandrosterone . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
2.1. Metabolism in the liver. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
2.1.1. DHEA metabolites. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
2.1.2. Epiandrosterone metabolites . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
2.2. Metabolism in the intestine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
2.2.1. DHEA metabolites. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
2.2.2. EpiA metabolites. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
2.3. Metabolism in the brain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
2.4. DHEA metabolites in cerebrospinal fluid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
3. Biological activities of DHEA and EpiA metabolites . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

Abbreviations: AD, Alzheimer’s disease; DHEA, 3b-hydroxy-androst-5-en-17-

one, dehydroepiandrosterone; EpiA, 3b-hydroxy-5a-androstan-17-one, epiandros-
terone; GC–MS, gas chromatography–mass spectrometry; LC–MS, liquid chroma-
tography–mass spectrometry; 3b,17b-AAD, 5a-androstane-3b,17b-diol; 3b,7a,17b-
AAT, 5a-androstane-3b,7a,17b-triol; 3b,7b,17b-AAT, 5a-androstane-3b,7b,17b-
triol; ADT, 5a-3b-hydroxy-androstan-17-one, androsterone; Adione, 5a-andro-
stane-3,17-dione; 4-adione, androst-4-ene-3,17-dione; 7a-HO-DHEA, 7a-hydroxy-
DHEA; 7b-HO-DHEA, 7b-hydroxy-DHEA; 3b,17a-AED, androst-5-ene 3b,17a diol;
3b,17b-AED, androst-5-ene-3b,17b-diol; 3b,7a,17b-AET, 5a-androst-5-ene-
3b,7a,17b-triol; 3b7b,17b-AET, 5a-androst-5ene-3b,7b,17b-triol; T, testosterone.
⇑ Tel.: +33 0 2 31 56 68 12; fax: +33 0 2 31 56 68 03.
E-mail address: laila.elkihel@unicaen.fr

0039-128X/$ - see front matter Ó 2011 Elsevier Inc. All rights reserved.
doi:10.1016/j.steroids.2011.09.008
 L.E. Kihel / Steroids 77 (2012) 10–26 11

3.1. DHEA metabolites and anticancer properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

3.2. DHEA metabolites and their effects on the brain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
3.2.1. DHEA metabolites and neuroprotective effects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
3.2.2. DHEA metabolites and memory . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
3.2.3. DHEA metabolites and neurodegenerative diseases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
3.3. DHEA metabolite effects on inflammatory and immune processes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
3.4. DHEA metabolites and osteoporosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
3.5. DHEA metabolites and thermogenesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
3.6. DHEA metabolites and adipogenesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
3.7. DHEA metabolites and the skin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
3.8. DHEA metabolites and the cardiovascular system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
3.9. DHEA metabolites and estrogenic receptors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
3.10. DHEA metabolites and androgenic receptors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
3.11. DHEA metabolites and the human prostate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
4. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

1. Introduction Clinical and experimental studies have shown that DHEA/

Dehydroepiandrosterone (3b-hydroxy-androst-5en-17-one,
DHEA) is an endogenous steroid synthesized in the adrenal cortex,
gonads, brain, and gastrointestinal tract. Its sulfated derivative,
DHEAS, is the most abundant circulating steroid in young adult
humans. Its levels decline dramatically with age and have been
correlated with degenerative changes related to aging. DHEA may
directly maintain bone, muscle, skin, and brain cells, modify the
immune response, inflammation, and lipid metabolism, and
ameliorate the deleterious effects of glucocorticoids [1–3].
DHEAS in humans and other mammals are multi-functional
steroids implicated in a broad range of biological processes, includ-
ing obesity [4], diabetes [5], bone metabolism [6], neuroprotection
[7], atherosclerosis [8], and anti-tumorigenesis [9]. DHEA and its
sulfate are also potent neuroactive steroids through their modula-
tory effects on the c-aminobutyric (GABA) and N-methyl-D-aspar-
tate (NMDA) receptors [10].
Moreover, DHEA is the physiological precursor in the synthesis
of androgens and estrogen via androst-4-ene-3,17-dione (4-adi-
one) in humans [11,12]. Many studies have been undertaken in

Oxygenated DHEA metabolites

O
O O

O O
HO HO S
HO -O
Pregnenolone O DHEA-sulfate
Cholesterol
Dehydroepiandrosterone (DHEA)
Other steroids
O
OH OH O

O
O O HO
H Androst-4-ene-3,17-dione (4-Adione) H
Dihydrotestosterone Testosterone
Etiocholanolone

OH
O hydroxylated 4-Adione metabolites

oxygenated Epi-A metabolites

HO HO
H H
5α-androstane-3β,17β−diol (3β,17β-AAD)
Epiandrosterone (EpiA)
Fig. 1. Biosynthetic pathway of DHEA and EpiA modified scheme from [21,22].
12 L.E. Kihel / Steroids 77 (2012) 10–26

O
OH

O
O O
OH S HO
-O O
DHEA-sulfate OH
Androst-5-ene-3β,17β-diol
HO
3β,16α-dihydroxy-androst-5-ene-17-one
O

O
O
OH O
HO
OH
O
Androst-4-ene-3,17-dione
HO O
HO
DHEA

O OH OH
O

O O
O O HO OH HO OH S OH
OH
-O
S
7α−ΟΗ−DHEA -O O
O

O O OH OH

O O
O O HO O O
O HO O S
S
-O O
7- oxo-DHEA -O O

O O OH OH

O O O
O HO OH HO OH OH
S OH S
-O O 7β−ΟΗ−DHEA -O O

Fig. 2. An overview of summary oxygenated DHEA metabolites identified after incubation of DHEA with rat liver homogenate Scheme slightly modified from Marwah et al.
[14]. See also [21,27,29].

attempts to elucidate the mechanism(s) responsible for these ef-
fects, but the data available so far indicate that DHEA acts at vari-
ous sites and levels rather than by a unifying mechanism. The
mechanism by which DHEA exerts its pleiotropic effects in humans
and rodents is not well understood but may involve metabolism of
DHEA into biologically active oxygenated metabolites [13,14].
2. Oxidative metabolism of DHEA and epiandrosterone
DHEA is a naturally occurring C-19 adrenal steroid that is
derived from cholesterol by a series of cytochrome P450 reactions,
and DHEA is a precursor for the metabolite epiandrosterone (3a-
hydroxy-5a-androstan-17-one, EpiA). It should be noted that EpiA
is formed by two mechanisms: (1) total enone ring A reduction of
androst-4-ene-3,17-dione (4-adione) and (2) selective reduction of
the D4 double bond and 3a-ketone of 4-adione to generate 5a-
androstane-3a,17a-diol (3a,17a-AAD), followed by 17a-oxidation
to generate EpiA (Fig. 1). DHEA is metabolized to androgens and
estrogens in steroidogenic tissues. It is also metabolized with EpiA
into oxygenated derivatives in mammalian tissues, such as the
intestine, brain, and especially human liver. The cytochrome
P450-7B1 (CYP7B1) uses NADPH and is responsible for the 7a-
hydroxylation of all 3b-hydroxysteroids (except cholesterol). The
3b-hydroxysteroids include pregnenolone and 5a-androstane-
3a,17-a-diol in tissues, such as the brain, liver, skin, and joints,
where the hydroxylase is expressed. These tissues also contain
NADP(H)-dependent 11b-hydroxysteroid dehydrogenase type 1
(11b-HSD1), which converts 7a-hydroxysteroids to the corre-
sponding 7b-hydroxysteroids [15–19]. Thus, DHEA metabolism is
the key to understanding several physiological events. DHEA
metabolism has also been studied in the brain, spleen, thymus,
perianal skin, ventral skin, intestine, colon, and muscle tissues
from mice [20].
Herein, we focus on the metabolism of DHEA and EpiA in the li-
ver, intestine, and brain.
2.1. Metabolism in the liver
The liver is one of the most important organs in the metabolism
of drugs [23]. Biotransformation involves phase I and phase II path-
ways and mainly takes place in the liver and intestine [24,25].
Phase I is usually oxidative, and these reactions are mainly cata-
lyzed by various enzymes of the Cytochrome P450 (CYP) supergene
family, which are predominantly expressed in the liver and play a
central role in drug metabolism. In phase II, such oxidized moieties
are conjugated with highly polar molecules, mainly glucuronic acid
 L.E. Kihel / Steroids 77 (2012) 10–26
or sulfate [26]. When these steroids are administered per os to
humans, the first organ encountered is the liver, where extensive
metabolism takes place. Most orally administrated steroids are
metabolized in the liver and excreted in the urine. As the liver is
responsible for xenobiotic (including steroids) elimination through
phase I (Cytochrome P450-mediated hydroxylation) and phase II
(conjugation) processes, it has been suggested that hydroxylated
steroid derivatives mainly protect against excessive steroid
production.
2.1.1. DHEA metabolites
DHEA metabolites have been studied extensively [14,27], and
many investigators have investigated the conversion of DHEA to
a variety of metabolites using a specific tissue fraction as the en-
zyme source [28–33]. Special attention has been paid to the study
of oxygenated DHEA metabolites [14]. The transformation of DHEA
was examined in whole rat liver homogenate in the presence of
malic acid using NADH and NADPH. The aim was to predict the
type of reactions that might occur in vivo, where the product of
one organelle may be the substrate for another, and to study the
timing of metabolite appearances to predict the sequence of their
formation. Twenty oxygenated DHEA metabolites were identified
and characterized using the highly specific and selective analysis
method of liquid chromatography–mass spectrometry (LC–MS).
These results showed that 7a-hydroxy-DHEA was the first product
formed from DHEA, which was converted via the sequence 7a-hy-
droxy-DHEA ? 7-oxo-DHEA ? 7a-hydroxy-DHEA. These deriva-
tives were also converted to diols, triols and sulfate esters (Fig. 2).
In the livers of rats, mice, and humans, 7a-hydroxylation of
DHEA is carried out by several P450 cytochromes (such as CYP7B1).
Additionally, one of the most abundant cytochromes in the liver is
the CYP3A forms CYP3A4 and CYP3A5, which have been reported
to carry out the 7a-hydroxylation of DHEA [13]. Fitzpatrick et al.
identified the DHEA metabolites formed by rodent and human liver
microsomal fractions in the presence of NADPH and O2. Hydroxyl-
ated metabolites, principally 7a-HO-DHEA, 7-oxo-DHEA, and 16a-
HO-DHEA, were identified in both species [13]. These authors and
others have demonstrated the conversion of 7a- and 7b-HO-DHEA
to 7-oxo-DHEA by rat and human isozymes of the 11b-hydroxy-
steroid dehydrogenases [34,35].
Another study identified and quantitated the liver microsomal
oxidative metabolism of DHEA by rodent, hamster, pig, and human
microsomal fractions (quantification of DHEA and metabolites by
O
17α-HSD
HO
DHEA
3β-HSD
O OH
17α-HSD
O O
Androst-4-ene-3,17-dione (4-Adione)
Fig. 3. Conversion of DHEA into Androst-5-ene-3b,17a-diol (3b,17a-AED) by the enzyme 17a-hydroxysteroid dehydrogenase (17a-HSD) [37].
13
gas chromatography–mass spectrometry (GC–Ms) analysis). In
adult liver microsomes, CYP3A4 and CYP3A5 were the P450 cyto-
chromes responsible for the production of 7a-hydroxy-DHEA,
16a-hydroxy-DHEA, and the previously unidentified hydroxylated
metabolite 7b-hydroxy-DHEA (reported by Fitzpatrick et al. [13]).
Whereas the fetal/neonatal form CYP3A7 produced 16a-hydroxy
and 7b-hydroxy-DHEA, CYP3A23 uniquely generated 7a-hydro-
xy-DHEA. Furthermore, other P450s, including CYP2B1, CYP2C11,
and CYP2D1, are responsible for 16a-hydroxy-DHEA metabolite
production in rat liver microsomal fractions [36]. Additionally, Ste-
vens et al. observed that CYP3A7 is expressed nearly exclusively in
human neonates [28]. The results of these studies demonstrate
that the regio- and stereo-selectivity of hydroxylation of DHEA
by specific P450s account for the unique DHEA metabolite profiles
formed by various species.
A recent report attempted to study the transformation of DHEA
and its 7- or 16a-hydroxylated derivatives in S9 fractions of human
liver microsomes with the cofactor NAD(H) or NADP(H) using a
radiolabeled steroid substrate for quantification and gas chroma-
tography–mass spectrometry (GC–MS) for identification [15]. The
results showed that DHEA was transformed into its 7- or 16a-
hydroxylated metabolites and 17b-hydroxylated derivatives in
the presence of NAD(P)H. 17b-Hydroxysteroid dehydrogenase
was responsible for the reduction of 17-oxo-steroids into 17b-
hydroxysteroids. The generation of 16a-hydroxy-DHEA from DHEA
indicated the expression of CYP3A4 and CYP3A5 in liver S9 frac-
tions. It was demonstrated that liver microsomal preparations
produce equal amounts of 7-oxo and 7b-hydroxy-DHEA from
7b-hydroxy-DHEA. It was also demonstrated that 7a/7b-hydroxy-
and 7-oxo-DHEA are not substrates for 16a-hydroxylase. In
addition, these data confirm the 7a/7b interconversion via a 7-
oxo-DHEA intermediate using an oxidoreduction process catalyzed
by 11a-hydroxysteroid dehydrogenase type 1. However, 17a-
hydroxysteroid dehydrogenase (17a-HSD) is responsible for the
reduction of 17-oxo-steroids into 17a-hydroxysteroids [37]. These
authors have shown that 17a-HSD converts DHEA to androst-5-
ene-3a,17a-diol (3a,17a-AED) (Fig. 3). To date, little information
is available about the biosynthesis, distribution, and biological
function of 3a,17a-AED.
11b-HSD1 has been reported to catalyze the interconversion of
several 7-oxygenated sterols and steroids. Muller et al. used micro-
somes from yeast expressing recombinant human 11b-HSD1 and
demonstrated the interconversion between 7-hydroxy- and
OH
HO
Androst-5-ene-3β,17α-diol (3β,17α−AED)
3β-HSD
17α,-hydroxy-androst-4-en-3-one (Epi-Testosterone)
14 L.E. Kihel / Steroids 77 (2012) 10–26

Androst-5-ene-3β,7α,16α,17β-tetrol Androst-5-ene-3α,7α,16α,17β-tetrol
OH OH OH OH

OH OH OH OH

I II III IV
HO OH HO OH HO OH HO OH

Androst-5-ene-3α,7β,16α,17β-tetrol
Androst-5-ene-3β,7β,16α,17β-tetrol
Fig. 4. Androst-5-enetetrols obtained by human liver microsomes identified by Ahlem et al. [39].

OH

HO
H

O O
O
OH
OH

HO HO
HO OH H H
HO OH H Epiandrosterone (EpiA)
H
7α-HO-EpiA
O
?
?
O O

O
? H

HO O HO OH
H H

7-Oxo-EpiA 7β-HO-EpiA
Not identified

Fig. 5. Summary EpiA oxygenated metabolites identified after EpiA incubation with human liver S9 fractions according to Chalbot et al. [15]. See also [21]. ?: Enzyme
pathways are not clearly identified.

7-oxo-DHEA. A high level of production of 7b-hydroxy-DHEA was
observed. The Km values obtained for the reduction of 7-oxo-DHEA
and the oxidation of 7b-hydroxy-DHEA were about half that for
cortisone and cortisol, respectively, indicating that 11b-HSD1 has
a higher affinity for the 7-oxo-DHEA metabolites than for glucocor-
ticoids [19].
Recent findings suggest preferential formation of 7b-hydroxy-
DHEA from 7-oxo-DHEA as well as some conversion of
7a-hydroxy-DHEA to 7b-hydroxy-DHEA. The equilibrium between
7-oxo- and 7-hydroxy-DHEA is regulated by 11bHSD1 and greatly
depends on the co-expression of hexose-6-phosphate dehydroge-
nase [32]. These findings identified 11b-HSD1 as the enzyme
responsible for the generation of various 7b-hydroxysteroid
metabolites [38]. Novel components of the human metabolome
were obtained by metabolic conversion of androst-5-ene precur-
sors to tetrols (I–IV) by human liver microsomes (Fig. 4). Of these
tetrols, the 7b-hydroxy epimers (I and III), but not the 7a-epimers,
were detected in human plasma and urine. This is the first report of

O OH

17β-hydroxysteroid oxido-reductase

HO R HO R

R = H, DHEA R = H, 3β-17β-AED
R = α-OH, 3β,7α,17β-AET
R = α-OH, 7α-HO-DHEA
R = β-OH, 3β,7β,17β-AET
R = β-OH, 7β-HO-DHEA
Fig. 6. Transformation of DHEA, 7a-hydroxy-DHEA and 7b-hydroxy-DHEA substrates in microsomes of human intestine. The steroid metabolites were identified by GC/MS
analysis. Scheme modified from Chalbot et al. [47].
 L.E. Kihel / Steroids 77 (2012) 10–26 15

a natural source of the androst-5-enetetrols 7b-epimers in human However, to the best of our knowledge, very few studies on
tissues [39]. DHEA, EpiA, and their metabolites have been carried out in the
intestine.

2.1.2. Epiandrosterone metabolites

2.2.1. DHEA metabolites
Androgenic steroids are derived from DHEA after puberty and
In one study, the [4–14C]-labeled steroid substrates (DHEA, 7a-
include epiandrosterone, which shares a closely related structure
hydroxy-DHEA and 7b-hydroxy-DHEA) were produced [46] by
with DHEA and is 7a-hydroxylated [16]. The 7a-hydroxylated
incubation of different cofactors with microsomes of human intes-
derivatives of EpiA may provide inactivation of androgen potencies
tine, and the metabolites were identified by gas chromatography-
and are of interest because of their promising biological effects
mass spectrometry [47]. The sole metabolites resulting from the
[40,41]. 7a-Hydroxy-EpiA is produced from EpiA via cytochrome
DHEA incubations were 17b-reduced derivatives, obtained by
P450-7B1 (CYP7B1) in tissues where the hydroxylase is expressed,
reduction of 17-ketone steroids. This enzymatic reduction process
including the brain, liver, skin and joints [16,18,42]. These tissues
implies the presence of a 17b-hydroxysteroid oxidoreductase. The
also contain NADP(H)-dependent 11b-HSD1, which converts 7a-
same finding applies to 7a-hydroxy-DHEA and 7b-hydroxy-DHEA
hydroxysteroids to the corresponding 7b-hydroxysteroids
metabolism, where 17b-reduction resulted in the respective triol
[19,42,43].
production (Fig. 6).
Investigations of EpiA metabolism in S9 fractions of the human
Except for 17b-reduced transformation, no other metabolites of
liver provided evidence for EpiA transformation to their 17b-
DHEA, 7a-hydroxy-DHEA, and 7b-hydroxy-DHEA have been ob-
hydroxylated derivatives, 7- or 16-a-hydroxylated metabolites
served. However, the formation of 17b-hydroxy-metabolites pro-
(Fig. 5). Provided that 16a-hydroxy-EpiA is generated by cyto-
duced by either the 17b-hydroxysteroid oxidoreductase or by
chrome P450 CYP3A4, a significantly larger amount of 16a-EpiA
other enzymes cannot be directly identified. These results also
was obtained in females than in males. 7a-Hydroxy-EpiA and 7b-
show that the tested intestine microsome fraction did not contain
hydroxy-EpiA interconvert to a degree, but no 7-oxo-EpiA interme-
the CYP7B1 responsible for the 7a-hydroxylation of DHEA. Neither
diate has been obtained, in contrast to the 7-hydroxylated DHEA
oxidation products of 7a- and 7b-hydroxy-DHEA nor inter-conver-
inter-conversion via 7-oxo-DHEA [15]. It can be postulated that
sion of these steroids were detected in the digests.
an unknown epimerase is involved in the inter-conversion process.
These data provided evidence that 11b-HSD1, depending on the
2.2.2. EpiA metabolites
NAD(H) or NADP(H) cofactors, can function as an epimerase that
Metabolism of EpiA, 7a-hydroxy-EpiA and 7b-hydroxy-EpiA
catalyzes the interconversion between 7a- and 7b-hydroxy-EpiA
was explored in the same manner by Chalbot and Morfin [47].
and between 5a-androstane-3b,7a,17b-triol (3b,7a,17b-AAT) and
The major metabolites of DHEA obtained were 17b-reduced deriv-
5a-androstane-3b,7b,17b-triol (3b,7b,17b-AAT), respectively, with
atives, and a 7a-hydroxylated metabolite of EpiA was not found
a preferential formation of the 7b-hydroxy metabolite [42,43].
(Fig. 7).
Additionally, no 5a,7-oxo-androstan-3b,17b-diol intermediate
In contrast, experiments with 7a-hydroxy-EpiA or 7b-hydroxy-
was present in the 5a-androstan-3b,7a,17b-triol/5a-androstan-
EpiA have provided evidence for the inter-conversion of these
3b,7b,17b-triol inter-conversion with a yeast-expressed
steroids. No trace of a 7-oxo derivative in this epimerization was
recombinant human 11bHSD1 [43]. Instead, interconversion of
detected. The identity of the enzyme responsible for the 7a-hydro-
the 7a- and 7b-hydroxylated forms was observed, with a prefer-
xy-EpiA/7b-hydroxy-EpiA inter-conversion is unknown, and to our
ence for the production of the 7b-hydroxylated epimers. Also, EpiA
knowledge, few studies have been carried out to investigate
was oxidized at the 3b-position; this is in contrast with DHEA.
whether an oxidoreductive process is used in this inter-conversion
Additional recent studies have not clearly indicated the presence
[21,42]. This is in contrast with results on 7a- and 7b-hydroxy-
of 7-oxo-EpiA as an intermediate in the conversion of 7a-EpiA to
DHEA, where no inter-conversion was observed. This makes the
7a-EpiA [21].
presence of an 11b-HSD1 in the human intestine doubtful.
However, it is possible that human intestine microsomes used
2.2. Metabolism in the intestine 7a- and 7b-hydroxy-EpiA and not the 7-hydroxy-DHEA epimers
as substrates for an unknown epimerase [47].
DHEA metabolism has been widely studied in the intestine and
other tissues (brain, spleen, thymus, skin, colon, coecum and mus- 2.3. Metabolism in the brain
cle) from mice. 7a-Hydroxy-DHEA and androst-5-ene-3b,17b-diol
were identified as DHEA metabolites in digests of all tissues DHEA and pregnenolone are formed in the brain independent of
[20,44,45]. the adrenal glands and gonads and are called neuro-steroids. While

O OH

17β-hydroxysteroid oxido-reductase

HO
H R HO R
H

R = H, EpiA R = H, 3β,17β-AAD
R = α-OH, 7α-HO-EpiA R = α-OH, 3β,7α,17β−ΑΑΤ
R = β-OH, 7β-HO-EpiA R = β-OH, 3β,7β,17β-AAT
Fig. 7. Transformation of EpiA, 7a-hydroxy-EpiA or 7b-hydroxy-EpiA substrates in microsomes of human intestine. The steroid metabolites were identified by GC/MS
analysis. Scheme modified from Chalbot et al. [47].
16 L.E. Kihel / Steroids 77 (2012) 10–26
DHEA is present at high levels in the blood under normal condi-
tions, it is found in the brain at even higher levels than in the
periphery.
The first step in steroid biosynthesis corresponds to the forma-
tion of pregnenolone from available cholesterol by oligodendro-
cytes in the rat brain, which are then converted to DHEA by the
cytochrome P450 enzyme 17a-hydroxylase (CYP17). This enzyme,
which is characteristic of all steroidogenic cells, has been shown to
be expressed in the nervous system of different classes of verte-
brates, including amphibians [48,49], birds [50], and mammals
[51–53]. However, the expression of this enzyme and its activity
are undetectable in the adult brain [54]. A recent study showed
that DHEA formation is independent of cytochrome P450 17a-
hydroxylase/C17,20-lyase activity in the mouse brain. This is the
first in vivo report demonstrating that there is a DHEA biosynthetic
pathway in brain tissue that is distinct from the CYP17-dependent
pathway present in peripheral steroidogenic tissues [55]. To date,
this controversy remains unresolved.
DHEA may be metabolized to active metabolites in the brain,
which is then responsible for some or all of the neuroprotective ef-
fects. Although the metabolic profile of DHEA in the brain has not
been fully elucidated, 7a-HO-DHEA and 7b-HO-DHEA are the most
abundant metabolites, and they play important roles in the central
nervous system [56].
Numerous studies have demonstrated that the adult rodent
brain is the site of extensive conversion of DHEA to 7a-hydroxy-
DHEA [56,57]. However, the metabolism of neuro-steroids, includ-
ing DHEA, in the adult human brain has been not extensively stud-
ied [58,59].
Cytochrome P4507B is principally expressed in the brain
[13,60] and has been found to be responsible for the 7a-hydroxyl-
ation of several 3b-hydroxysteroids, including DHEA, and EpiA.
One study describes the in vitro conversion of DHEA into both
O
O O
OH S
-O
HO
3β,16α-dihydroxyandrost-5-ene-17-one
HO
OH
HO
Androst-5-ene-3β,17α-diol
HO
not identified
Fig. 8. Summary oxygenated DHEA metabolites identified in the brain. ?: Enzyme pathways are not clearly identified.
7a-HO-DHEA and 3b,17b-AED in the aging brains of healthy hu-
man subjects. The structures of the brain that were studied were
the frontal cortex, hippocampus, amygdala, cerebellum and stria-
tum. The synthesis of 7a-HO-DHEA was found in all of the brain re-
gions evaluated. The amount of 7a-HO-DHEA formed in the frontal
cortex was significantly higher compared with that found in all of
the other brain regions [61]. The synthesis of 3b,17b-AED was also
observed in all the human brain structures studied. However, a sig-
nificant difference was observed between the regions. The cerebel-
lum and striatum had significantly higher conversion rates of
DHEA to 3b,17b-AED compared to the frontal cortex, hippocampus
and amygdala. The formation of 3b,17b-AED from DHEA in the ro-
dent brain has also been documented [57]. 7a-HO-DHEA, 7b-HO-
DHEA, and 16a-HO-DHEA are derived from DHEA in the human
brain, where 7a-hydroxylation and 16a-hydroxylation are cata-
lyzed by CYP7B1 and CYP3A7, respectively [62,63]. The 5-andos-
tenediols (3a17a-AED and 3a, 17b-AED) are produced by the
metabolism of DHEA by 17a-hydroxysteroid dehydrogenase or
17b-hydroxysteroid dehydrogenase, respectively [37,64–66].
3b,17a-AED and 3b,17b-AED are neuro-steroids produced in neu-
ro-ectodermal tissue by the metabolism of DHEA, and they exist
in the a- and b-epimeric forms [67]. It has been suggested that
3b,17b-AED and 17b-hydroxysteroid dehydrogenase, which are
mainly expressed in astrocytes and microglia, may play a neuro-
protective role in the brain [68,69]. 17a-Hydroxysteroid dehydro-
genase is also expressed in the central nervous system, as well as
other organs, but the biological function of 3b,17a-AED has not
been clearly identified [37,68].
A recent study reported a detailed analysis of steroids in the
adult male rat brain using GC–MS and applying the identification
criteria of retention time and diagnostic ion ratios at a high level
of stringency. Among the steroids identified were DHEA and EpiA,
which was detected in two-ion Selected Ion Monitoring (SIM, in
OH
O
DHEA-sulfate
HO
Androst-5-ene-3β,17β-diol
O
O
HO OH
7α−ΗΟ−DHEA
DHEA ?
?
?
O
O
?
HO O
7- oxo-DHEA
OH
not identified
7β−ΗΟ−DHEA
 L.E. Kihel / Steroids 77 (2012) 10–26 17

Table 1
Focus on recent finding of oxygenated DHEA metabolite activities.

DHEA metabolites Biological effect References

7oxo-DHEA, 7a-HO-DHEA, 7b-HO-DHEA, Anticancer, cell lines Hep G2, Caco-2, HT-29 [29]
Androsterone, EpiA, Etiocholanolone
3b,17a-AED, 3b,17b-AED Anticancer cell lines: Basal growth of murine RAW 264.7, P388D1, and human HL-60 cells, human [83–
U937 lyphomaT98G and U251 MG gioblastoma, T98G, U87MG, U251MG, LN-18, LN-229 and LN-Z308 85,87]
glioma cell lines
3b,17a-AED, 3b,7a,17a-AET, 3b, 7b,17a-AET, induced apoptosis in U937 lymphoma cells and autophagy in T98G [88]
3b,7a,17b-AET
Effects on the brain
7-oxo-DHEA acetate Beneficial effect for memory in old mice by antagonizing the GABAA receptor [96]
7a-HO-EpiA and 7b-HO-EpiA, 7b-HO-EpiA protection against hypoxia-induced neuronal damage, neuroprotective in two in vivo rat models of [40]
cerebral ischaemia-induced neurodegeneration
7b-HO-EpiA neuroprotective effect in animal models of Alzheimer’s disease [41]
Effects in inflammatory and in immune process
7a-HO-DHEA rheumatoid arthritis: 7a-OH-DHEA levels is 5-fold higher than nonarthritic mice [116–118]
7b-HO-EpiA Anti-oxidant and anti-inflammatory: reduced prostaglandin E2 (PGE2) production [112]
7a-HO-DHEA High antioxidant effect in the colon than DHEA, protective effect against sodium dextran sulfate- [110,111]
induced colitis in rats
7b-HO-EpiA regulation of cytoprotective prostaglandins from human mononuclear cells [113]
7a-HO-DHEA, 7b-HO-DHEA and 7b-HO-EpiA Regulation of the prostaglandin synthesis pathway in human peripheral blood monocytes [114]
3b, 7b,17b-AET limit the production of autoimmune Th1 associated cytokines, and ultimately may be beneficial for [120]
patients with multiple sclerosis or other autoimmune diseases
3b, 7b,17b-AET improves survival in a rodent model of traumatic shock [122]
Androst-5-ene-3a,7b,16a,17b-tetrol, Androst-5- Anti-inflammatory effects in rodent disease models of multiple sclerosis, lung injury, chronic [39]
ene-3b,7b,16a,17b-tetrol prostatitis, and colitis
Osteoporosis
3b,17a-AED, 3b,17b-AED, 3b,7b,17b-AET, 3b,17a-AED promotes bone resorption while 3b,17a-AED inhibits it [132]
3b,7b,17b-AET
3b, 7b,17b-AET preserves bone mineral content, limits cortical cancellous bone loss and preserves endochondral [134]
growth rate
Thermogenesis
7-oxo-DHEA Effect on temporarily actual levels of thyroid hormones [143]
7-oxo-DHEA and 7-oxo-DHEA acetate Thermomodulating agent [137,142]
7a- and 7b-HO-DHEA 7b-HO-DHEA levels, decreased in patients with manifest hypothyroidism (negative relationship) [144]
Adipogenesis
DHEA, 7-oxo-DHEA, 7b-HO-DHEA, 7b-HO- DHEA act as a thermogenic agent in adipocytes, but 7-oxo-DHEA appears to act as a nonthermogenic [147]
DHEA, 3b, 7a,17b-AET, 3b, 7b,17b-AET enhancer of differentiation
Cardiovascular system
3b,17b-AED improves cardiovascular function and organ blood flow [159,160]

mass spectrometry) of the sulfate fraction but not confirmed with
three-ion SIM. This inconsistency is probably related to the analyt-
ical method used [70]. This work did not identify DHEA and EpiA
oxygenated metabolites, and these results differ from several pre-
vious reports [71,72] that detected pregnenolone sulfate in rat
brain.
More recently, in another study, both 7a-hydroxy and 7b-hy-
droxy-DHEA were generated by incubation of DHEA with rat brain
microsomes; 7a-hydroxy-DHEA was the major metabolite [73].
Investigators have demonstrated that 11beta-HSD1 may convert
7a-HO-DHEA to 7b-HO-DHEA in the liver and other tissues
[17,74,75], but whether the same mechanism for the conversion
from 7a-HO-DHEA into 7b-HO-DHEA also occurs in the brain is
not yet known; future studies should focus on clearly identifying
the enzyme(s) responsible for the conversion of 7a-HO-DHEA to
7b-HO-DHEA [59]. It should be noted that no unknown P450 cyto-
chrome has been shown to be responsible for the direct 7beta-
hydroxylation of DHEA [76]. To the best of our knowledge, there
are currently no data demonstrating the presence of 7-oxo-DHEA
in the brain (Fig. 8).
2.4. DHEA metabolites in cerebrospinal fluid
Levels of 7a-hydroxy-DHEA, 7b-hydroxy-DHEA, and 16a-hy-
droxy-DHEA in cerebrospinal fluid (CSF) have been reported, and
the levels of these neuro-steroids in CSF may be helpful in the diag-
nosis of neurodegenerative diseases and in differentiating Alzhei-
mer’s disease (AD) from vascular dementia [77]. It is currently
unknown whether CSF neuro-steroid levels are related to brain
neuro-steroid levels in humans. Recent findings indicate that the
CSF level of DHEA is positively correlated with the temporal cortex
level and that CSF DHEA is elevated in AD and positively correlates
with neuropathological disease stage [78]. Another report on DHEA
derivatives in the human ventricular CSF identified 7a-HO-DHEA,
7b-HO-DHEA, 3b,7a,17b-AET, and 3b,7b,17b-AET. However, the en-
try site of DHEA metabolites into ventricular CSF and their role in
this compartment are not known [79].
3. Biological activities of DHEA and EpiA metabolites
7-Oxygenated steroids are widespread in mammals, birds, fish,
and plants [67]. The significant bioactivity of DHEA and its metab-
olites has been noted in previous studies. This is relevant particu-
larly for the 7a- and 7b-hydroxylated metabolites of DHEA and
other 17-oxosteroids [40,80,81]. DHEA and its metabolites are
responsible for the above-mentioned immunoprotective and
neuroprotective effects. However, evidence has accumulated that
7-oxygenated DHEA derivatives may act as potent local immuno-
protective agents, which are in fact responsible for many immuno-
modulatory and antiglucocorticoid effects previously ascribed to
DHEA [80]. Thus, most of the functions initially attributed to DHEA
from observations in rodents are now thought to be due to these
oxygenated metabolites.
As DHEA is a precursor for the metabolite EpiA, we present here
a single paragraph on the biological properties of DHEA and EpiA
metabolites. A summary of recent findings on oxygenated DHEA
metabolites is presented in Table 1.
18 L.E. Kihel / Steroids 77 (2012) 10–26
3.1. DHEA metabolites and anticancer properties
7-Oxygenated DHEA derivatives (7oxo-DHEA, 7a-HO-DHEA,
and 7b-HO-DHEA), androsterone, epiandrosterone, and etiocholan-
olone have anti-proliferative effects on human hepatocellular liver
carcinoma cells (Hep G2), human epithelial colorectal adenocarci-
noma cells (Caco 2), and human colon adenocarcinoma grade II
cells (HT-29). Whether the pentose phosphate pathway (glucose-
6-phosphate dehydrogenase (G6PD), [82]) or the mevalonate path-
way (3-hydroxy-3-methylglutaryl-CoA reductase (HMGR)), which
are involved in cholesterol biosynthesis, or the mitogen-activated
protein kinase (MAPK)-mediated signal transduction pathway for
DNA synthesis were more responsible for the growth inhibition
was investigated [29]. Because the growth inhibitory effects were
not correlated with the inhibition of G6PD nor HMGR activity,
some other mechanism for the growth inhibition may exist. How-
ever, these data do not exclude the possibility that the anti-prolif-
erative effects of any one of the individual steroids may have been
mediated via the inhibition of G6PD and/or HMGR. These non-
androgenic DHEA metabolites may serve as chemopreventive or
anti-proliferative therapies.
3b,17a-AED and 3b,17b-AED are neuro-steroids produced in
neuro-ectodermal tissue by the metabolism of DHEA. The biologi-
cal actions of these two epimers are distinct. 3b,17a-AED can in-
duce apoptosis in human lymphoma cells and autophagic cell
death in malignant glioma cells, and the antiproliferative functions
of this epimer are not dependent on either estrogen or androgen
receptors [83–85]. In contrast, 3b,17b-AED enhances the immune
system with minimal anti-tumor activity [86]. More recently, one
study indicated that autophagy in human malignant glioma cells
and transformation of fibroblasts by treatment with the neuro-ste-
roid 3b,17a-AED proceeds through an endoplasmic reticulum
stress-dependent mechanism accompanied by partial activation
of the unfolded protein response. Moreover, these results implicate
PERK/eIF2a signaling as one of the pathways involved in 3b,17a-
AED-mediated autophagy and cell death [87]. Through the same
pathway, 3b,17a-AED and 3b,7a,17a-AET induce apoptosis in
U937 lymphoma cells and autophagy in T98G glioblastoma cells.
These results indicate that the position of the hydroxyl group on
C-17 in the a-configuration dictates the anti-tumor activity of
the androstene. The addition of a third hydroxyl in the a-position
on C-7 did not increase the antitumor activity of 3a,17a-AED but
rather reduced the potency by approximately two-fold. These find-
ings demonstrate a strict structure–function relationship in the
anti-tumor activity for androstenediol. Moreover, 3a,17a-AED
and 3a,7a,17a-AET retain their abilities to induce autophagic cell
death in glial tumors, and apoptosis prevails in lymphomas, indi-
cating that the mechanism of cell death for these two steroids is
dependent upon the origin of the tumor tissue [88].
3.2. DHEA metabolites and their effects on the brain
3.2.1. DHEA metabolites and neuroprotective effects
Neuroprotective effects of DHEA metabolites have been docu-
mented both in vitro and in vivo. In the central nervous system,
3a,17b-AED is believed to play a neuroprotective role, but a biolog-
ical function for 3b,17a-AED has not been identified [68]. It has
been suggested that the 7-hydroxylated metabolites of DHEA
should be considered as possible mediators of some of the neuro-
protective effects of DHEA in the brain [89]. 7a- and 7a-HO-DHEA
have neuroprotective potency mediated by antiglucocorticoid
activity, which once again confirmed the hypothesis that the 7-
hydroxylation process may explain the neuroprotection offered
by neuro-steroids [56].
Recent reports indicate that DHEA metabolites are the local
agents responsible for the neuroprotective effects. This is relevant
particularly for the 7-oxygenated metabolites of DHEA and other
17-oxosteroids [40,81]. A study by Pringle et al. demonstrated that
7b-HO-EpiA can protect against neuronal death in both in vivo and
in vitro models of reperfusion ischemia-induced damage. However,
it is unclear how these 7-hydroxy steroids prevent cell damage
[41].
The rate of aromatization of 4-adione and 7-hydroxylation of
DHEA by different neuronal cell lines from fetal rat and mouse
brains was compared to that of embryonic rat hippocampal cells
in primary culture. The comparison of the metabolism of DHEA
and 4-adione by neural cells of different origins shed light on the
relative importance of the aromatization and 7-hydroxylation
pathways in the brain and their roles in neuroprotection [90,91].
Numerous studies have reported neuroprotective effects of
DHEA metabolites on the nervous system, even in aged animals,
but the results of these studies are not conclusive. To date, the
in vivo effects of DHEA and its metabolites in the brain have not
been fully elucidated, and the possibility that these oxygenated
derivatives of DHEA might be products involved in neuroprotec-
tion needs to be investigated further.
3.2.2. DHEA metabolites and memory
Studies have been carried out to assess the neuroprotective ef-
fects of DHEA and its metabolites and their ability to enhance
memory and learning in humans and animals [89,92–95].
7-oxo-DHEA has been shown to reverse scopolamine-induced
memory impairment in young mice [96]. A significant and selec-
tive decrease in hippocampal CYP7B bioactivity was demonstrated
in 24-month-old rats with impaired spatial memory performance
in the water maze but not in age-matched cognitively intact rats
[97]. Further, treatment of cognitively impaired aged rats with
the CYP7B product 7a-hydroxypregnenolone was shown to en-
hance spatial memory retention after a 30 min delay in the radial
arm water maze. In contrast, the parent steroid pregnenolone
had no significant effect at the dose tested. These authors suggest
that if 7a-hydroxylated metabolites are more active than the par-
ent steroid, this may explain why oral DHEA replacement in the el-
derly has not successfully improved memory [98,99] while DHEA
administration is effective at improving memory and mood in
healthy young subjects [100], as young subjects have greater hip-
pocampal CYP7B activity. Perhaps the 7a-hydroxylated metabolite
rather than the parent steroid would be a more effective therapy in
the elderly.
3.2.3. DHEA metabolites and neurodegenerative diseases
Dementia, a hallmark of the aging central nervous system, has
been partially attributed to a lack of circulating steroids, such as
DHEA, in several studies [101]. The pathogenesis of age-associated
disorders, such as neurodegenerative diseases, with a particular
focus on Alzheimer’s disease (AD), and their connection with DHEA
and its metabolites will be discussed here. The most frequent cause
of dementia is AD (more than 60%) [102], which is a progressive
neurodegenerative disorder with gradual onset and deterioration
of cognitive functions, such as memory, language and visuo-spatial
skills [103].
Hormone replacement therapy with DHEA is a frequently dis-
cussed topic, especially in AD [1]. DHEA is a well-known inhibitor
of glucose 6-phosphate dehydrogenase (G6PD), and it is hypothe-
sized that reduced G6PDH activity has a beneficial effect on age-
related disease development and longevity [104]. Because NADPH
is a key cofactor in the activity of many antioxidative and reductive
enzymes [105], its depletion may result in an impairment of fuel
utilization and consequently the onset of the disease. In addition
to its other roles, NADPH is a co-factor for the enzyme CYP7B1,
which is abundant in brain tissues and responsible for
 L.E. Kihel / Steroids 77 (2012) 10–26
7a-hydroxylation of DHEA [106]. The expression of this enzyme in
post-mortem brain tissues is decreased in AD [107].
The metabolism of DHEA was recently analyzed in the aging
brain from Alzheimer patients and non-demented controls. The
conversion of DHEA to 3b,17b-AED and to 7a-OH-DHEA occurs in
the frontal cortex, hippocampus, amygdala, cerebellum and stria-
tum of both Alzheimer’s patients and controls. The formation of
these metabolites within distinct brain regions is negatively corre-
lated with the density of b-amyloid deposits [108].
Epiandrosterone is also converted to 7b-OH-EpiA in numerous
tissues. Indeed, DHEA and epiandrosterone (EpiA), which is derived
from DHEA, are ineffective in protecting organotypic hippocampal
slice cultures against hypoxia/reperfusion-induced neuronal cell
death, whereas their corresponding 7-hydroxy derivatives had
powerful neuroprotective effects [40]. Therefore, 7a-hydroxy
DHEA and 7b-hydroxy EpiA may be endogenous neuroprotective
agents, and 7-hydroxylation of DHEA and EpiA may be impaired
in neurodegenerative conditions, such as AD.
Other findings indicate that 7b-HO-DHEA has powerful cytopro-
tective effects, suggesting that this neuro-steroid may have thera-
peutic potential in various neurodegenerative conditions, such as
AD, and 7b-hydroxy steroids may constitute a novel class of endog-
enous neuroprotective agents [41]. It was also reported that
3b,17b-AED may regulate the cerebral levels of DHEA and may con-
tribute to the control of DHEA activity in the aging brain and in Alz-
heimer’s disease [61,109].
DHEA and DHEAS have been widely studied in neurodegenera-
tive diseases, but the true role of oxygenated DHEA metabolites in
these diseases is still not clear. Several reports on DHEA metabo-
lites have suggested their neuroprotective effects, but these have
not been proven. Further research is necessary to clarify this topic.
3.3. DHEA metabolite effects on inflammatory and immune processes
The antioxidant effects of DHEA and 7a-hydroxy-DHEA against
oxidative stress induced by colitis were investigated in vivo in rats.
DHEA and 7a-hydroxy-DHEA exerted a significant anti-oxidant ef-
fect against oxidative stress induced colitis in rats, reducing oxida-
tive damage to proteins and lipids. These findings demonstrate
that DHEA does not act alone and that its 7-hydroxylated metabo-
lites may be of importance for its protective effects [110,111].
Strong anti-oxidant and anti-inflammatory properties of 7b-hy-
droxy-EpiA have been reported in vivo in rats. Although all dose
levels of 7b-hydroxy-EpiA reduced prostaglandin E2 (PGE2) pro-
duction, only the lowest dose (0.01 mg/kg) completely prevented
colitis damage and tissue inflammation. 7b-Hydroxy-EpiA pre-
treatment prevents the occurrence of sodium dextran sulfate-in-
duced colitis through a shift from PGE2 to PGD2 production [112].
A more recent study demonstrated an important role for 7b-hy-
droxy-EpiA in the regulation of cytoprotective prostaglandins from
human mononuclear cells. An elevation in the circulating levels of
these prostanoids may be important in the neuroprotective actions
of 7b-hydroxy-EpiA [113].
The effects of 7a-hydroxy-DHEA, 7b-hydroxy-DHEA and 7b-hy-
droxy-EpiA on prostaglandin (PG) production and related enzyme
gene expression were tested. Human peripheral blood monocytes
were activated by incubation with the inflammatory cytokine
TNF-a in the presence and absence of these hydroxysteroids. The
results suggest that 7b-hydroxy-EpiA produced locally at the site
of inflammation may trigger a shift in the PG pathway, favoring in-
creased production of 15d-PGJ2 at the expense of PGE2. The pro-
tective effects of 15d-PGJ2 on neighboring cells could contribute
to the resolution of inflammation and tissue repair, thus facilitating
the immune response [114].
The conversion of DHEA to downstream steroid hormones in
target macrophages, as major effector cells in rheumatoid arthritis
19
(RA), has also been investigated [115]. Within 1 day of culture with
radiolabeled DHEA, monocyte-derived macrophages converted
DHEA to D5-derivatives, such as 16a-hydroxy-DHEA, 3b,17b-
AED, and 3b, 16a, 17b-AET. The conversion of DHEA leads to in-
creased downstream effector hormones in target macrophages,
which may be an important factor for local immunomodulation.
The conversion of DHEA to downstream effector hormones and
the regulation of key enzymes in macrophages may explain some
of the effects of DHEA therapy in chronic inflammatory diseases
or arteriosclerosis. A study on DHEA metabolism in arthritis
showed that the severity of arthritis correlates with increased lev-
els of the P450 enzyme CYP7B, which converts DHEA into7a-hy-
droxy-DHEA [116]. Increased CYP7B activity leads to higher
levels of the DHEA metabolite hydroxy-DHEA in synovial fluid,
which may contribute to the maintenance of the chronic inflam-
mation observed in rheumatoid arthritis patients [117,118].
3b,7b,17b-AET was described previously to be a more highly
oxygenated natural DHEA metabolite found in humans [119] that
provides anti-inflammatory benefits in animal models of autoim-
munity and trauma [120–125].
In rodents, DHEA is metabolized to produce (among other com-
pounds) 3b,7b,17b-AET, which has potent anti-inflammatory activ-
ity. Its functions in the body may include tissue-specific
modulation of glucocorticoid action, immune function, and control
of acute and chronic inflammation [122,123]. The anti-inflamma-
tory activity of two tetrols (androst-5-ene-3b,7b,16a,17b-tetrol
(I) and androst-5-ene-3a,7b,16a,17b-tetrol (III, Fig. 4) was demon-
strated in rodent models of acute and chronic inflammation. Anti-
inflammatory properties were observed with exogenous adminis-
tration of these compounds in rodent disease models of multiple
sclerosis, lung injury, chronic prostatitis, and colitis [39].
3.4. DHEA metabolites and osteoporosis
In rodents, DHEA opposes certain activities of endogenous glu-
cocorticoids, such as osteoporosis [126,127], and improves osteo-
blast growth and bone tissue morphometry [128]. As the
literature grew, DHEA became widely used as an anti-aging and
anti-stress dietary supplement and was considered a promising
agent to treat osteoporosis. Despite promising results in rodents,
DHEA provided only modest improvement in bone mineral density
(BMD) in patients treated with GC and elicited unwanted andro-
genic side effects [129–131].
The discovery of the receptor activator of nuclear factor-jB li-
gand (RANKL)/RANK/osteoprotegerin system and its role in the
regulation of bone resorption exemplifies how both serendipity
and a logic-based approach can identify factors that regulate cell
function.
The study examined the particular effects of a and b androst-5-
ene steroid epimers (3b,17b-AED, 3b,7b,17b-AET, 3b,17a-AED, and
3b,7a,17b-AET) on immortalized human fetal osteoblasts in cul-
ture and examined the ability of a/b androstenediol metabolites
to signal effects by regulating PPAR in osteoblasts. The results
showed that 3b,17a-AED opposes 3b,17b-AED by elevating bone
resorption in osteoblast cells. The increase in RANKL/OPG is mod-
ulated by the activation of PPAR-c that in turn caused apoptosis of
FOB-9 cells [132].
The potential role of 3b,7b,17b-AET in bone formation is high-
lighted by these in vitro findings related to 3b,7b,17b-AET-induced
OPG expression in MG-63 osteoblasts treated with dexametha-
sone, a synthetic glucocorticoid. OPG is a soluble decoy RANK
receptor that is secreted by osteoblasts and binds to the surface
of RANK ligand (RANKL) on osteoblasts. This decreases the pool
of RANKL available for RANK receptor binding, decreasing the acti-
vation of the RANK receptor on pre-osteoclasts and ultimately
osteoclast commitment and differentiation [133].
20 L.E. Kihel / Steroids 77 (2012) 10–26
Recent findings showed that 3b,7b,17b-AET preserves bone
mineral content, but not mineral density, from thermal trauma-in-
duced bone loss, and it limits cortical cancellous bone loss and pre-
serves the endochondral growth rate but not cortical bone loss
[134].
3.5. DHEA metabolites and thermogenesis
Thyroid hormones are the main hormonal regulators of
thermogenesis.
Feeding DHEA to rats enhances heat production and decreases
the efficiency of food utilization. The effects of DHEA were mea-
sured by the production of liver mitochondrial sn-glycerol-3-phos-
phate dehydrogenase and cytosolic malic enzyme, two enzymes
that had been demonstrated to be induced by the classic thermo-
genic agent thyroid hormone [135,136].
Derivatives of DHEA were investigated for their ergogenic prop-
erties, and 7-oxo-DHEA in particular was shown to be 4-fold more
thermogenic than DHEA [137]. Both 7-hydroxy-DHEA epimers and
7-oxo-DHEA are more thermogenic than DHEA itself. The thermo-
genic properties of DHEA and 7-oxo-DHEA are likely to be due to
two interrelated actions: increased levels of enzymes and proteins
that shuttle substrate and electrons in and out of the mitochondria
and an increased proton leak across the mitochondrial inner mem-
brane [138]. There is greater experimental evidence for increased
shuttling by DHEA and 7-oxo-DHEA than for induction of a proton
leak. The levels of malic enzyme, which converts pyruvate to ma-
late, can be increased more than 5-fold by 7-oxo-DHEA [139]. In
further experiments, mitochondria from rats treated with 7-oxo-
DHEA appear to have an increased proton leak across the inner
mitochondrial membrane, similar to thyroid hormone [140]. Fur-
thermore, plasma levels of DHEA and its metabolites, such as
3b,17a-AED, may be altered in thyroid disorders [141].
Previous studies have attempted to establish a relationship be-
tween thyroid laboratory parameters, including major thyroid
auto-antibodies and DHEA and its 7-hydroxylated metabolites,
which are believed to act as immunoprotective agents [80,142].
Recently, it was demonstrated that administration of 7-oxo-DHEA,
one of the candidates of steroid replacement therapy, to healthy
male volunteers temporarily influenced the actual levels of thyroid
hormones [143].
Other data clearly demonstrates a negative relationship be-
tween the levels of the immunoprotective steroids (7a-HO- and
7b-HO-DHEA) and the extent of the autoimmune process, at least
in hypothyroidism [144].
3.6. DHEA metabolites and adipogenesis
Adipocytes have been intensively studied in recent years, and
the mouse embryo-derived 3T3-L1 cell line has often been used
as a model system for the study of preadipocyte differentiation,
maturation, and cellular processes [145].
Adipose tissue contains significant quantities of DHEA (10-
times greater than in the circulation) and possesses the appropri-
ate enzymatic machinery for the metabolism of DHEA. Thus, inves-
tigators have examined a role for DHEA in adipose tissue biology,
and despite the low circulating levels of DHEA/DHEAS in rodents,
many have used murine cell lines or in vivo rodent models as their
experimental systems. DHEA has been reported to inhibit 3T3-L1
adipogenesis and proliferation [146].
The anti-adipogenic effect of DHEA on human omental pre-adi-
pocytes in vitro is in agreement with the significant decrease in
abdominal fat in elderly subjects treated with DHEA for 6 months
[4].
The effect of DHEA and 7-oxygenated metabolites (7-oxo-DHEA,
7a-OH-DHEA, 7b-OH-DHEA, androst-5ene-3b, 7b,17b-triol and
androst-5ene-3b,7a,17b-triol) on the expression, accumulation,
and catalytic activity of key lipid-metabolizing enzymes in 3T3-
L1 cells as the experimental model was examined [147]. This work
showed that 7-oxo-DHEA has an opposite effect on adipocyte dif-
ferentiation and thermogenesis. These disparate effects suggest
that DHEA and 7-oxo-DHEA may be targeting different aspects of
the differentiation program.
Another study identified and quantitated DHEA metabolites
in adipose cells (3T3-L1) by LC–MS analysis. These results
demonstrated that adipocytes rapidly converted DHEA to and-
rost-5-ene-3b,17b-diol. 7a-Hydroxy-DHEA was interconverted
with 7-oxo-DHEA and 7b-hydroxy-DHEA and the corresponding
17b reduced products (Fig. 9). DHEA and its derivatives were
detected only in the culture medium, suggesting that DHEA is
metabolized via peripheral or integral cell membrane enzymes.
Alternatively, there may be efficient mechanisms at play for the
secretion of the steroids to the aqueous media rather than reten-
tion in the lipid-rich cell. Interestingly, the authors identified
3b,17b-AED as the major metabolite of DHEA [148].
3.7. DHEA metabolites and the skin
Similar to the classical steroidogenic organs, such as gonads and
adrenal glands, the skin and its appendages, including hair follicles,
sebaceous glands, and eccrine/apocrine glands, are armed with all
of the necessary enzymes required for androgen synthesis and
metabolism [149]. However, after cessation of estrogen secretion
by the ovaries at menopause, practically all sex steroids acting in
the skin are synthesized locally from DHEA [150]. Immunohisto-
chemical studies have demonstrated the presence of both CYP7B1
and 11beta-HSD1 in human skin and other tissues [17].
Some effects of DHEA on human skin, including collagen gene
expression and protein synthesis, have already been described
[151–155]. For example, recent findings strongly suggest the pos-
sibility that DHEA exerts anti-aging effects in the skin through
stimulation of collagen biosynthesis and improved structural orga-
nization of the dermis while modulating keratinocyte metabolism
[154].
The major product of DHEA metabolism in EpiSkin is DHEAS
(88%), while the other metabolites are 7a-HO-DHEA (8.2%), 4-adi-
one (1.3%), 3b,17b-AED (1.3%), dihydrotestosterone (DHT) (1.4%)
and androsterone (ADT) (2.3%) [153]. However, the role of oxygen-
ated DHEA metabolites in the prevention of skin aging has not been
clearly established.
3.8. DHEA metabolites and the cardiovascular system
Previous work has shown that left ventricular performance, car-
diac output, and organ blood flow in the liver, small intestine, and
kidney decreased significantly after trauma-induced hemorrhage
[156–158].
One study indicated that 3b,17b-AED administration after trau-
ma-induced hemorrhage improves cardiovascular function and or-
gan blood flow and decreases cytokine production in male animals.
This improvement of cardiovascular function may be associated
with attenuation in IL-6 levels and an increase in plasma nitrite/ni-
trate levels. Because 3b,17b-AED produced the above-mentioned
effect, 3b,17b-AED appears to be a useful adjuvant for restoring de-
pressed cardiovascular function and attenuating cytokine produc-
tion after trauma-induced hemorrhage [159,160]. To the best of
our knowledge, only the effect of 3b,17b-AED has been studied.
3.9. DHEA metabolites and estrogenic receptors
Estrogens exert a number of biological effects throughout the
body, most of which are mediated via the estrogen receptors ERa
 L.E. Kihel / Steroids 77 (2012) 10–26
O
HO
HO
3β,17β−AED
DHEA
O
HO OH
HO
7α-HO-DHEA
3β,7α,17α−AET
O O
HO O HO
7-oxo-DHEA 7β-HO-DHEA
Fig. 9. Proposed metabolic route for metabolism of DHEA in differentiating 3T3-L1 adipocytes. Modified scheme from Marwah et al. [148].
and b. Estrogens play important functions in the female as well as
the male reproductive system and in several non-reproductive tis-
sues. For instance, it has been proposed that ERa might stimulate
proliferation whereas ERb inhibits proliferation [161,162]. DHEA
and its metabolites have been evaluated as ligand activators of
ERa and ERb [163,164]. In some studies, it has been proposed that
activation of ERb by 5a-androstane-3b,17b-diol (3b,17b-AAD) may
be important for suppression of prostate growth [165].
Because 3b,17b-AAD is metabolized by CYP7B1, it has been sug-
gested that this enzyme might decrease ERb action via control of
3b,17b-AAD levels. In contrast, other investigators have reported
that a steroid metabolite formed by CYP7B1, 7a-hydroxy-DHEA,
activates ERb and that CYP7B1 catalysis therefore should increase
ERb action [166]. Furthermore, a cholesterol derivative metabo-
lized by CYP7B1, 27-hydroxycholesterol, was recently found to ex-
ert pro-estrogenic actions in some cells but suppress ER activation
in others [167,168]. Recent data indicate that low concentrations of
3b,17b-AED can trigger ER-mediated responses equally well for
both ERa and b and that CYP7B1-mediated conversion of 3b,17b-
AED into a 7a-hydroxy-metabolite will result in a loss of activity
[169].
3.10. DHEA metabolites and androgenic receptors
A ligand sensitive androgen receptor (AR) is involved in the reg-
ulation of prostate growth, muscle and bone mass, and spermato-
genesis in males. AR agonists are therapeutically useful in the
treatment of osteoporosis, cachexia, contraception, and androgen
deficiency [168,170], and antagonists are used for the treatment
of prostate cancer. It has been shown that 3b,17b-AED is a strong
activator of AR target genes, but DHEA and 7-oxo-DHEA cannot
activate AR target genes in the absence or presence of the co-acti-
vator ARA70 in DU145 cells co-transfected with a reporter gene
and expression plasmid (wtAR, mtAR877, or mtAR708) [171–
21
OH O
HO
Testosterone
OH
4-Adione
OH
OH
HO OH
OH
3β,7β,17β−AET
173]. Other studies have suggested that when there is low metab-
olism of DHEA or if 4-adione and 3b,17b-AED are produced, andro-
genic effects are observed, while other metabolites (DHEA-S,
7a-HO-DHEA, 7b-HO-DHEA, and 7-oxo-DHEA) essentially lack
androgenic activities [174].
3.11. DHEA metabolites and the human prostate
The prostate is an androgen-dependent organ, the growth,
maintenance and secretory functions of which are stimulated by
the continuous presence of androgens and growth factors. In hu-
mans, the adrenal glands secrete a large amount of the precursor
steroid DHEA, which can be converted into androst-4-ene-3,17-
dione or androst-5-ene-3b,17b-diol and then testosterone (T) and
further metabolized into the potent androgen 5a-dihydrotestos-
terone in prostatic tissues [175].
Various steroidogenic enzymes in the prostate are involved in
the biosynthesis and metabolism of steroid precursors, such as
DHEA [176], and steroidogenesis could differ in different species,
such as human, dog, rat and mouse prostate tissues, as well as pri-
mary cells and prostatic cancer cell lines [177]. The in vitro DHEA
metabolites formed from human prostate homogenates were
identified by using LC–MS and GC–MS techniques. Androst-
5-ene-3b,17b-diol was the main DHEA metabolite identified.
7a-hydroxy-DHEA, 7a-hydroxy-DHEA, 7-oxo-DHEA, androst-4-
ene-3,17-dione, testosterone, and 5a-dihydrotestosterone were
identified as well [178].
Androgens are involved in many diseases, especially benign
prostatic hyperplasia and prostate cancer. However, few reports
have investigated the levels of adrenal androgens in prostate can-
cer tissues after androgen deprivation [179].
In recent studies, analyses of steroids were performed using LC–
MS in castration-resistant prostate cancer tissues after androgen
deprivation therapy. The levels of DHEA, androst-4-ene-3,
22 L.E. Kihel / Steroids 77 (2012) 10–26
17-dione, androst-5-ene-3b,17b-diol, testosterone and dihydrotes-
tosterone in prostate cancer were detected by using LC–MS/MS for
a precise simultaneous quantitative analysis. They found that the
levels of DHEA were high in prostate cancer tissues, irrespective
of androgen deprivation therapy. These authors suggested that
DHEA plays a significant role in the synthesis of T and DHT in pros-
tate cancer tissues after androgen deprivation therapy [180].
It should be noted that androst-5-ene-3b,17b-diol is found at a
high concentration in prostate cancer tissue, even after androgen
deprivation therapy. Indeed, its androgen activity is not inhibited
by the antiandrogens currently used to treat prostate cancer pa-
tients [173].
DHEA can directly activate the androgen receptor (AR) or estro-
gen receptor beta (ERb) in the prostatic epithelium or the AR or
estrogen receptor alpha (ERa) in the prostatic stroma. Indirect ef-
fects are conferred by DHEA metabolism of androgenic ligands
(including androstenedione, testosterone and dihydrotestosterone)
or estrogenic ligands (including 7-OH-DHEA, 3b-adiol, or 17b-
estradiol) [181]. Alternatively, DHEA metabolites may act on ERb
in the prostate, potentially antagonizing androgenic effects on
prostate cancer growth, such as DHEA metabolism into 7a-hydro-
xy-DHEA (7HD), a known ligand for ERb [182]. The complexity of
the balance between androgenic and estrogenic effects, whether
as direct ligands or DHEA metabolites on the prostate, is matched
by the complexity of estrogen action through ERa and ERb. The
intratissular balance of androgen and estrogen levels is most
important for prostatic development and differentiation [183–
185]. Many questions remain to be clarified on the regulation of
DHEA metabolism in the prostate. Oxidative metabolism of DHEA
has not been fully investigated either in primary cells or prostatic
cancer cell lines [186–189]. This unresolved question remains:
what biochemical mechanism promotes the androgenic and estro-
genic metabolism of DHEA?
4. Conclusion
In recent years, significant interest has arisen from the hypoth-
eses that declining DHEA concentrations in adults may serve as an
indicator of a number of pathological conditions associated with
aging, many of which are correlated with changes in lipid metabo-
lism. However, the precise role of DHEA and its metabolites re-
mains uncertain in spite of many epidemiological and biological
studies. Many of the functions initially attributed to DHEA from
observations in humans and animals are now thought to be prop-
erties of its oxygenated metabolites. However, the genomic effects
of DHEA and its major metabolites remain controversial because
the metabolism of DHEA varies in different cells and tissues. Recent
studies have described DHEA metabolism in different tissues, such
as in rodent and human livers, and identification of an extensive
number of oxygenated DHEA metabolites has been achieved. How-
ever, the major metabolic pathway for DHEA in extra-hepatic tis-
sues has not fully been elucidated. Indeed, the study of the
biochemical pathway of DHEA metabolites in the brain is an attrac-
tive alternative for exploring the pathways involved in the patho-
genesis of neurodegenerative diseases.
Further characterization of the physiological and molecular
mechanisms of DHEA and its metabolites would help to clarify
their potential roles in cellular metabolism. Moreover, elucidating
the molecular mechanisms of DHEA and its oxygenated metabo-
lites in preventing the initiation of aging of the brain and periphe-
ral tissues would offer immense opportunities for the development
of future treatments.
For instance, the diversity of highly oxygenated steroids identi-
fied in the human liver raises the possibility of naturally occurring
diols, triols and tetrols and perhaps even more highly oxygenated
species with as yet undescribed chemical and biological activities.
Once considered to be physiologically inactive, these species repre-
sent a novel pharmacological target for designing new DHEA ana-
logs with potent biological activity that might be devoid of
unwanted estrogenic or androgenic side effects. These data should
be explored for the development of a specific synthetic derivative
that is less labile to metabolism and may at last deliver the benefits
of oxygenated DHEA metabolites observed in animals and humans.
Thus, synthetic derivatives of oxygenated DHEA metabolites could
be used as a possible platform to derive novel pharmaceutical
products.
References
[1] Cameron DR, Braunstein GD. The use of dehydroepiandrosterone therapy in
clinical practice. Treat Endocrinol 2005;4(2):95–114.
[2] Bovenberg SA, van Uum SH, Hermus AR. Dehydroepiandrosterone
administration in humans: evidence based? Neth J Med 2005;63(8):300–4.
[3] Oberbeck R, Kobbe P. Dehydroepiandrosterone (DHEA): a steroid with
multiple effects. Is there any possible option in the treatment of critical
illness? Curr Med Chem 2010;17(11):1039–47.
[4] Villareal DT, Holloszy JO. Effect of DHEA on abdominal fat and insulin action
in elderly women and men: a randomized controlled trial. J Am Med Assoc
2004;292(18):2243–8.
[5] Talaei A, Amini M, Siavash M, Zare M. The effect of dehydroepiandrosterone
on insulin resistance in patients with impaired glucose tolerance. Hormones
2010;9(4):326–31.
[6] Tok EC, Ertunc D, Oz U, Camdeviren H, Ozdemir G, Dilek S. The effect of
circulating androgens on bone mineral density in postmenopausal women.
Maturitas 2004;48(3):235–42.
[7] Juhasz-Vedres G, Rozsa E, Rakos G, et al. Dehydroepiandrosterone sulfate is
neuroprotective when administered either before or after injury in a focal
cortical cold lesion model. Endocrinology 2006;147:683–6.
[8] Wang L, Hao Q, Wang YD, Wang WJ, Li DJ. Protective effects of
dehydroepiandrosterone on atherosclerosis in ovariectomized rabbits via
alleviating inflammatory injury in endothelial cells. Atherosclerosis
2011;214(1):47–57.
[9] Celec P, Starka L. Dehydroepiandrosterone - is the fountain of youth drying
out? Physiol Res 2003;52:397–407.
[10] Zheng P. Neuroactive steroid regulation of neurotransmitter release in the
CNS: action, mechanism and possible significance. Prog Neurobiol
2009;89(2):134–52.
[11] Labrie F, Luu-the V, Labrie C, Simard J. DHEA and its transformation into
androgens and estrogens in peripheral target tissues: Intracrinology. Front
Neuroendocrinol 2001;22:185–212.
[12] Labrie F. DHEA, important source of sex steroids in men and even more in
women. Prog Brain Res 2010;182:97–148.
[13] Fitzpatrick JL, Ripp SL, Smith NB, Pierce WM, Prough RA. Metabolism of DHEA
by cytochromes P450 in rat and human liver microsomal fractions. Arch
Biochem Biophys 2001;389:278–87.
[14] Marwah A, Marwah P, Lardy H, Ergosteroids VI. Metabolism of
dehydroepiandrosterone by rat liver in vitro: a liquid chromatographic-
mass spectrometric study. J Chromatogr B Analyt Technol Biomed Life Sci
2002;767(2):285–99.
[15] Chalbot S, Morfin R. Human liver S9 fractions: metabolism of
dehydroepiandrosterone, epiandrosterone, and related 7-hydroxylated
derivatives. Drug Metab Dispos 2005;33:563–9.
[16] Kim SB, Chalbot S, Pompon D, Jo DH, Morfin R. The human cytochrome
P4507B1: catalytic activity studies. J Steroid Biochem Mol Biol
2004;92:383–9.
[17] Hennebert O, Chalbot S, Alran S, Morfin R. Dehydroepiandrosterone 7alpha-
hydroxylation in human tissues: possible interference with type 1 11beta-
hydroxysteroid dehydrogenase-mediated processes. J Steroid Biochem Mol
Biol 2007;104:326–33.
[18] Trap C, Nato F, Chalbot S, Kim SB, Lafaye P, Morfin R. Immunohistochemical
detection of the human cytochrome P4507B1: production of a monoclonal
antibody after cDNA immunization. J Neuroimmunol 2005;159:41–7.
[19] Muller C, Pompon D, Urban P, Morfin R. Inter-conversion of 7alpha- and
7beta-hydroxy-dehydroepiandrosterone by the human 11beta-
hydroxysteroid dehydrogenase type 1. J Steroid Biochem Mol Biol
2006;99:215–22.
[20] Morfin R, Courchay G. Pregnenolone and dehydroepiandrosterone as
precursors of native 7-hydroxylated metabolites which increase the
immune response in mice. J Steroid Biochem Mol Biol 1994;50(1–2):91–100.
[21] Hennebert O, Montes M, Favre-Reguillon A, Chermette H, Ferroud C, Morfin R.
Epimerase activity of the human 11beta-hydroxysteroid dehydrogenase type
1 on 7-hydroxylated C19-steroids. J Steroid Biochem Mol Biol 2009;114(1–
2):57–63.
[22] Lootens L, Van Eenoo P, Meuleman P, Leroux-Roels G, Delbeke FT. The uPA(+/
+)-SCID mouse with humanized liver as a model for in vivo metabolism of 4-
androstene-3, 17-dione. Drug Metab Dispos 2009;37(12):2367–74.
 L.E. Kihel / Steroids 77 (2012) 10–26
[23] Katoh M, Tateno C, Yoshizato K, Yokoi T. Chimeric mice with humanized liver.
Toxicology 2008;246(1):9–17.
[24] Kerkhof EG, Graaf IA, Jager MH, Groothuis GM. Induction of phase I and II drug
metabolism in rat small intestine and colon in vitro. Drug Metab Dispos
2007;35:898–907.
[25] Kerkhof EG, Ungell AB, Sjoberg AK, Jager MH, Hilgendorf C, Graaf IAM,
Groothuis GMM. Innovative methods to study human intestinal drug
metabolism in vitro: precision-cut slices compared with Ussing Chamber
preparations. Drug Metab Dispos 2006;34:1893–902.
[26] Wang S, Rijk JC, Riethoff-Poortman JH, Van Kuijk S, Peijnenburg AA, Bovee TF.
Bovine liver slices combined with an androgen transcriptional activation
assay: an in-vitro model to study the metabolism and bioactivity of steroids.
Anal Bioanal Chem 2010;397(2):631–41.
[27] Lardy H, Marwah A, Marwah P. Transformations of DHEA and its metabolites
by rat liver. Lipids 2002;37(12):1187–91.
[28] Stevens JC, Hines RN, Gu C, Koukouritaki SB, Manro JR, Tandler PJ, Zaya MJ.
Developmental expression of the major human hepatic CYP3A enzymes. J
Pharmacol Exp Ther 2003;307:573–82.
[29] Yoshida S, Honda A, Matsuzaki Y, Fukushima S, Tanaka N, Takagiwa A, et al.
Anti-proliferative action of endogenous dehydroepiandrosterone metabolites
on human cancer cell lines. Steroids 2003;68(1):73–83.
[30] Robinzon B, Miller KK, Prough RA. Biosynthesis of [3H]7 alpha-hydroxy-, 7
beta-hydroxy-, and 7-oxo-dehydroepiandrosterone using pig liver
microsomal fractions. Anal Biochem 2004;333(1):128–35.
[31] Baker ME. Co-evolution of steroidogenic, steroid-inactivating enzymes,
adrenal, sex steroid receptors. Mol cell Endocrinol 2004;215(1–2):55–62.
[32] Nashev LG, Chandsawangbhuwana C, Balazs Z, Atanasov AG, Dick B, Frey FJ,
Baker ME, Odermatt A. Hexose-6-phosphate dehydrogenase modulates
11beta-hydroxysteroid dehydrogenase type 1-dependent metabolism of 7-
keto- and 7beta-hydroxy-neurosteroids. PLoS One 2007;2(6):e561.
[33] Rijk JCW, Bovee TFH, Groot MJ, Peijnenburg AACM, Nielen MWF. Evidence of
the indirect hormonal activity of prohormones using liver S9 metabolic
bioactivation and an androgen bioassay. Anal Bioanal Chem
2008;392(3):417–25.
[34] Robinzon B, Michael KK, Ripp SL, Stephen J, Winters SJ, Prough RA.
Glucocorticoids inhibit interconversion of 7-hydroxy- and 7-oxo-
metabolites of dehydroepiandrosterone (DHEA): a role for 11b-
hydroxysteroid dehydrogenases? Arch Biochem Biophys 2003;412(2):251–8.
[35] Robinzon B, Prough RA. Interactions between dehydroepiandrosterone and
glucocorticoid metabolism in pig kidney: nuclear and microsomal 11beta-
hydroxysteroid dehydrogenases. Arch Biochem Biophys 2005;442(1):33–40.
[36] Miller KK, Cai J, Ripp SL, Pierce Jr WM, Rushmore TH, Prough RA. Stereo- and
regioselectivity account for the diversity of dehydroepiandrosterone (DHEA)
metabolites produced by liver microsomal cytochromes P450. Drug Metab
Dispos 2004;32(3):305–13.
[37] Bellemare V, Faucher F, Breton R, Luu-the V. Characterization of 17alpha-
hydroxysteroid dehydrogenase activity (17alpha-HSD) and its involvement in
the biosynthesis of epitestosterone. BMC Biochem 2005;6:12.
[38] Odermatt A, Nashev LG. The glucocorticoid-activating enzyme 11beta-
hydroxysteroid dehydrogenase type 1 has broad substrate specificity:
Physiological and toxicological considerations. J Steroid Biochem Mol Biol
2010;119(1–2):1–13.
[39] Ahlem CN, Page TM, Auci DL, Kennedy MR, Mangano K, Nicoletti F, Ge Y,
Huang Y, White SK, Villegas S, Conrad D, Wang A, Reading CL, Frincke JM.
Novel components of the human metabolome: the identification,
characterization and anti-inflammatory activity of two 5-androstene
tetrols. Steroids 2011;76(1–2):145–55.
[40] Pringle A, Schmidt W, Deans J, Wulfert E, Reymann K, Sundstrom L. 7-
Hydroxylated epiandrosterone (7-OH-EPIA) reduces ischemia-induced
neuronal damage both in vivo and in vitro. Eur J Neurosci 2003;18:117–24.
[41] Dudas B, Hanin I, Rose M, Wulfert E. Protection against inflammatory
neurodegeneration and glial cell death by 7beta-hydroxy epiandrosterone, a
novel neurosteroid. Neurobiol Dis 2004;15:262–8.
[42] Hennebert O, Pernelle C, Ferroud C, Morfin R. 7Alpha- and 7beta-hydroxy-
epiandrosterone as substrates and inhibitors for the human 11beta-
hydroxysteroid dehydrogenase type 1. J Steroid Biochem Mol Biol
2007;105:159–65.
[43] Hennebert O, Le Mee S, Pernelle C, Morfin R. 5Alpha-androstane-3beta,
7alpha, 17beta-triol and 5alpha-androstane-3beta, 7beta, 17beta-triol as
substrates for the human 11beta-hydroxysteroid dehydrogenase type 1.
Steroids 2007;72(13):855–64.
[44] Akwa Y, Morfin R, Robel P, Baulieu EE. Neurosteroids metabolism: 7a-
hydroxylation of dehydroepiandrosterone and pregnenolone by rat brain
microsomes. Biochem J 1992;288:959–64.
[45] Morfin R, Courchay G. Pregnenolone and dehydroepiandrosterone as
precursors of active 7-hydroxylated metabolites which increase the
immune response in mice. J Steroid Biochem Mol Biol 1994;50:91–100.
[46] Chalbot S, Trap C, Monin JP, Morfin R. Use of bioconversion for the
preparation of [4–14C]-labeled 7alpha- and 7beta-hydroxylated derivatives
of dehydroepiandrosterone and epiandrosterone. Steroids 2002;67:1121–7.
[47] Chalbot S, Morfin R. Neurosteroids: metabolism in human intestine
microsomes. Steroids 2005;70:319–26.
[48] Inai Y, Nagai K, Ukena K, Oishi T, Tsutsui K. Seasonal changes in neurosteroid
concentrations in the amphibian brain and environmental factors regulating
their changes. Brain Res 2003;959:214–25.
23
[49] Do Rego JL, Tremblay Y, Luu-The V, Repetto E, Castel H, Vallarino M, et al.
Immunohistochemical localization and biological activity of the
steroidogenic enzyme cytochrome P450 17alpha-hydroxylase/C17, 20-lyase
(P450C17) in the frog brain and pituitary. J Neurochem 2007;100(1):251–68.
[50] Tsutsui K, Matsunaga M, Miyabara H, Ukena K. Neurosteroid biosynthesis in
the quail brain: a review. J Exp Zool A Comp Exp Biol 2006;305(9):733–42.
[51] Schumacher M, Akwa Y, Guennoun R, Robert F, Labombarda F, Desarnaud F,
et al. Steroid synthesis and metabolism in the nervous system: trophic and
protective effects. J Neurocytol 2000;29(5–6):307–26.
[52] Mellon SH, Griffin LD. Neurosteroids: biochemistry and clinical significance.
Trends Endocrinol Metab 2002;13:35–43.
[53] Hojo Y, Hattori TA, Enami T, Furukawa A, Suzuki K, Ishii HT, Mukai H,
Morrison JH, Janssen WG, Kominami S, Harada N, Kimoto T, Kawato S. Adult
male rat hippocampus synthesizes estradiol from pregnenolone by
cytochromes P45017a and P450 aromatase localized in neurons. Proc. Natl
Acad Sci USA 2004;101:865–70.
[54] Cascio C, Brown RC, Hales DB, Papadopoulos V. Pathways of
dehydroepiandrosterone formation in rat brain glia. J Steroid Biochem Mol
Biol 2001;75:177–86.
[55] Liu Y, Pocivavsek A, Papadopoulos V. Dehydroepiandrosterone formation is
independent of cytochrome P450 17alpha-hydroxylase/17, 20 lyase activity
in the mouse brain. J Steroid Biochem Mol Biol 2009;115(3–5):86–90.
[56] Morfin R, Starka L. Neurosteroid 7-hydroxylation products in the brain. Int
Rev Neurobiol 2001;46:79–95.
[57] Steckelbroeck S, Watzka M, Lütjohann D, Makiola P, Nassen A, Hans VHJ, et al.
Characterization of the dehydroepiandrosterone (DHEA) metabolism via
oxysterol 7a-hydroxylase and 17-ketosteroid reductase activity in the human
brain. J Neurochem 2002;83:713–26.
[58] Stoffel-Wagner B. Neurosteroid metabolism in the human brain. Eur J
Endocrinol 2001;145:669–79.
[59] Li A, Bigelow JC. The 7-hydroxylation of dehydroepiandrosterone in rat brain.
Steroids 2010;75(6):404–10.
[60] Rose KA, Stapleton G, Dott K, Kieny MP, Best R, Schwarz M, et al. Cyp7b, a
novel brain cytochrome P450, catalyzes the synthesis of neurosteroids 7a-
hydroxy dehydroepiandrosterone and 7a-hydroxy pregnenolone. Proc Natl
Acad Sci USA 1997;94:4925–30.
[61] Weill-Engerer S, David JP, Sazdovitch V, Liere P, Schumacher M, Delacourte A,
et al. In vitro metabolism of dehydroepiandrosterone (DHEA) to 7alpha-
hydroxy-DHEA and Delta5-androstene-3beta, 17beta-diol in specific regions
of the aging brain from Alzheimer’s and non-demented patients. Brain Res
2003;969(1–2):117–25.
[62] Brown RC, Cascio C, Papadopoulos V. Neurosteroids: oxidative stress-
mediated dehydroepiandrosterone formation in Alzheimer’s disease
pathology. Neurobiol Aging 2000;21:S238.
[63] Yang HY, Lee QP, Rettie AE, Juchau MR. Functional cytochrome P4503A
isoforms in human embryonic tissues: expression during organogenesis. Mol
Pharmacol 1994;46:922–8.
[64] Steckelbroeck S, Watzka M, Reissinger A, Wegener-Toper P, Bidlingmaier F,
Bliesener N, Hans VH, Clusmann H, Ludwig M, Siekmann L, Klingmüller D.
Characterisation of estrogenic 17beta-hydroxysteroid dehydrogenase
(17beta-HSD) activity in the human brain. J Steroid Biochem Mol Biol
2003;86(1):79–92.
[65] Mindnich R, Möller G, Adamski J. The role of 17 beta-hydroxysteroid
dehydrogenases. Mol Cell Endocrinol 2004;218(1–2):7–20.
[66] Mindnich R, Deluca D, Adamski J. Identification and characterization of 17
beta-hydroxysteroid dehydrogenases in the zebrafish, Danio rerio. Mol Cell
Endocrinol 2004;215(1–2):19–30.
[67] Lathe R. Steroid and sterol 7-hydroxylation: ancient pathways. Steroids
2002;67:967–77.
[68] Zwain IH, Yen SS. Neurosteroidogenesis in astrocytes, oligodendrocytes, and
neurons of cerebral cortex of rat brain. Endocrinology 1999;140:3843–52.
[69] Gottfried-Blackmore A, Sierra A, Jellinck PH, McEwen BS, Bulloch K. Brain
microglia express steroid-converting enzymes in the mouse. J Steroid
Biochem Mol Biol 2008;109(1–2):96–107.
[70] Ebner MJ, Corol DI, Havlíková H, Honour JW, Fry JP. Identification of
neuroactive steroids and their precursors and metabolites in adult male rat
brain. Endocrinology 2006;147(1):179–90.
[71] Mathur C, Prasad VV, Raju VS, Welch M, Lieberman S. Steroids and their
conjugates in the mammalian brain. Proc Natl Acad Sci USA 1993;90:85–8.
[72] Liere P, Akwa Y, Weill Engerer S, Eychenne B, Pianos A, Robel P, et al.
Validation of an analytical procedure to measure trace amounts of
neurosteroids in brain tissue by gas chromatography–mass spectrometry. J
Chromatogr B Biomed Sci Appl 2000;739:301–12.
[73] Li A, May MP, Bigelow JC. An LC/MS method for the quantitative
determination of 7alpha-OH DHEA and 7beta-OH DHEA: an application for
the study of the metabolism of DHEA in rat brain. Biomed Chromatogr
2010;24(8):833–7.
[74] Tomlinson JW, Bujalska I, Stewart PM, Cooper MS. The role of 11 beta-
hydroxysteroid dehydrogenase in central obesity and osteoporosis. Endocr
Res 2000;26(4):711–22.
[75] Muller C, Hennebert O, Morfin R. The native anti-glucocorticoid paradigm. J
Steroid Biochem Mol Biol 2006;100:95–105.
[76] Doostzadeh J, Urban P, Pompon D, Morfin R. Pregnenolone-7 beta-
hydroxylating activities of yeast-expressed mouse cytochrome P450-1A1
and mouse-tissue microsomes. Eur J Biochem 1996;242(3):641–7.
24 L.E. Kihel / Steroids 77 (2012) 10–26
[77] Kim SB, Hill M, Kwak YT, Hampl R, Jo DH, Morfin R. Neurosteroids:
cerebrospinal fluid levels for Alzheimer’s disease and vascular dementia
diagnostics. J Clin Endocrinol Metab 2003;88:5199–206.
[78] Naylor JC, Hulette CM, Steffens DC, Shampine LJ, Ervin JF, Payne VM, et al.
Cerebrospinal fluid dehydroepiandrosterone levels are correlated with brain
dehydroepiandrosterone levels, elevated in Alzheimer’s disease, and related
to neuropathological disease stage. J Clin Endocrinol Metab
2008;93(8):3173–8.
[79] Starka L, Hill M, Kancheva R, Novak Z, Chrastina J, Pohanka M, et al. 7-
Hydroxylated derivatives of dehydroepiandrosterone in the human
ventricular cerebrospinal fluid. Neuro Endocrinol Lett 2009;30(3):368–72.
[80] Morfin R. Involvement of steroids and cytochromes P450 species in the
triggering of immune defense. J Steroid Biochem Mol Biol 2002;80:273–90.
[81] Trincal M, Loeper D, Pompon D, Hauw JJ, Morfin R. DHEA metabolism in the
brain production and effects of the 7a-hydroxylated derivative. In: Morfin R,
editor, DHEA and the brain. London: E-publishing Taylor and Francis; 2002.
pp. 117–128.
[82] Gordon G, Mackow MC, Levy HR. On the mechanism of interaction of steroids
with human glucose 6-phosphate dehydrogenase. Arch Biochem Biophys
1995;318:25–9.
[83] Huynh PN, Loria RM. Contrasting effects of alpha- and beta-androstenediol on
oncogenic myeloid cell lines in vitro. J Leukoc Biol 1997;62:258–67.
[84] Graf MR, Jia W, Loria RM. The neuro-steroid, 3beta androstene 17alpha diol
exhibits potent anti-tumour effects on human malignant glioma and
lymphoma cells through different programmed cell death pathways. Br J
Cancer 2007;97:619–27.
[85] Graf MR, Jia W, Johnson RS, Dent P, Mitchell C, Loria RM. Autophagy and the
functional roles of Atg5 and beclin-1 in the anti-tumor effects of 3b-
androstene 17a-diol neuro-steroid on malignant glioma cells. J Steroid
Biochem Mol Biol 2009;115(3–5):137–45.
[86] Loria RM. Immune up-regulation and tumor apoptosis by androstene
steroids. Steroids 2002;67:953–66.
[87] Jia W, Loria RM, Park MA, Yacoub A, Dent P, Graf MR. The neuro-steroid, 5-
androstene 3b, 17a diol; induces endoplasmic reticulum stress and
autophagy through PERK/eIF2a signaling in malignant glioma cells and
transformed fibroblasts. Int J Biochem Cell Biol 2010;42(12):2019–29.
[88] Graf MR, Jia W, Lewbart ML, Loria RM. The anti-tumor effects of androstene
steroids exhibit a strict structure-activity relationship dependent upon the
orientation of the hydroxyl group on carbon-17. Chem Biol Drug Des
2009;74(6):625–9.
[89] Jellinck PH, Lee SJ, McEwen BS. Metabolism of dehydroepiandrosterone by rat
hippocampal cells in culture: possible role of aromatization and 7-
hydroxylation in neuroprotection. J Steroid Biochem Mol Biol
2001;78(4):313–7.
[90] Jellinck PH, Croft G, McEwen BS, Gottfried-Blackmore A, Jones G, Byford V,
Bulloch K. Metabolism of dehydroepiandrosterone by rodent brain cell lines:
relationship between 7-hydroxylation and aromatization. J Steroid Biochem
Mol Biol 2005;93(1):81–6.
[91] Jellinck PH, Kaufmann M, Gottfried-Blackmore A, Croft G, Byford V, McEwen
BS, Jones G, Bulloch K. Dehydroepiandrosterone (DHEA) metabolism in the
brain: identification by liquid chromatography/mass spectrometry of the
delta-4-isomer of DHEA and related steroids formed from androstenedione
by mouse BV2 microglia. J Steroid Biochem Mol Biol 2006;98(1):41–7.
[92] Migues PV, Johnston AN, Rose SP. Dehydroepiandosterone and its sulphate
enhance memory retention in day-old chicks. Neuroscience
2002;109:243–51.
[93] Jellinck PH, Kaufmann M, Gottfried-Blackmore A, McEwen BS, Jones G,
Bulloch K. Selective conversion by microglia of dehydroepiandrosterone to 5-
androstenediol-A steroid with inherent estrogenic properties. J Steroid
Biochem Mol Biol 2007;107(3–5):156–62.
[94] Sujkovic E, Mileusnic R, Fry JP, Rose SP. Temporal effects of
dehydroepiandrosterone sulfate on memory formation in day-old chicks.
Neuroscience 2007;148(2):375–84.
[95] Aldred S, Mecocci P. Decreased dehydroepiandrosterone (DHEA) and
dehydroepiandrosterone sulfate (DHEAS) concentrations in plasma of
Alzheimer’s disease (AD) patients. Arch Gerontol Geriatr 2010;51(1):e16–8.
[96] Shi J, Schulze S, Lardy HA. The effect of 7-oxo-DHEA acetate on memory in
young and old C57BL/6 mice. Steroids 2000;65(3):124–9.
[97] Yau JL, Noble J, Graham M, Seckl JR. Central administration of a cytochrome
P450–7B product 7 alpha-hydroxypregnenolone improves spatial memory
retention in cognitively impaired aged rats. J Neurosci 2006;26(43):
11034–40.
[98] Wolf OT, Neumann O, Hellhammer DH, Geiben AC, Strasburger CJ,
Dressendorfer RA, et al. Effects of a two-week physiological
dehydroepiandrosterone substitution on cognitive performance and well-
being in healthy elderly women and men. J Clin Endocrinol Metab
1997;82:2363–7.
[99] Yamada S, Akishita M, Fukai S, Ogawa S, Yamaguchi K, Matsuyama J, Kozaki K,
Toba K, Ouchi Y. Effects of dehydroepiandrosterone supplementation on
cognitive function and activities of daily living in older women with mild to
moderate cognitive impairment. Geriatr Gerontol Int 2010;10(4):280–7.
[100] Alhaj HA, Massey AE, McAllister-Williams RH. Effects of DHEA administration
on episodic memory, cortisol and mood in healthy young men: a double-blind,
placebo-controlled study. Psychopharmacology (Berl) 2006;188(4):541–51.
[101] Makrantonaki E, Schönknecht P, Hossini AM, Kaiser E, Katsouli MM, Adjaye J,
Schröder J, Zouboulis CC. Skin and brain age together: the role of hormones in
the ageing process. Exp Gerontol 2010;45(10):801–13.
[102] Qiu C, De Ronchi D, Fratiglioni L. The epidemiology of the dementias: an
update. Curr Opin Psychiatry 2007;20:380–5.
[103] Schonknecht P, Pantel J, Kruse A, Schroder J. Prevalence and natural course of
aging-associated cognitive decline in a population-based sample of young-
old subjects. Am J psychiatry 2005;162(11):2071–7.
[104] Schwartz AG, Pashko LL. Dehydroepiandrosterone, glucose-6-phosphate
dehydrogenase, and longevity. Ageing Res Rev 2004;3:171–87.
[105] Mamelak M. Alzheimer’s disease, oxidative stress and
gammahydroxybutyrate. Neurobiol Aging 2007;28:1340–60.
[106] Chalbot S, Morfin R. Dehydroepiandrosterone metabolites and their
interactions in humans. Drug Metabol Drug Interact. 2006;22:1–23.
[107] Yau JL, Rasmuson S, Andrew R, Graham M, Noble J, Olsson T, et al.
Dehydroepiandrosterone 7-hydroxylase CYP7B: predominant expression in
primate hippocampus and reduced expression in Alzheimer disease.
Neuroscience 2003;121:307–14.
[108] Schumacher M, Weill-Engerer S, Liere P, Robert F, Franklin RJ, Garcia-Segura
LM, et al. Steroid hormones and neurosteroids in normal and pathological
aging of the nervous system. Prog Neurobiol 2003;71(1):3–29.
[109] Tagawa N, Sugimoto Y, Yamada J, Kobayashi Y. Strain differences of
neurosteroid levels in mouse brain. Steroids 2006;71(9):776–84.
[110] Pélissier MA, Trap C, Malewiak MI, Morfin R. Antioxidant effects of
dehydroepiandrosterone and 7a-hydroxy-dehydroepiandrosterone in the
rat colon, intestine and liver. Steroids 2004;69(2):137–44.
[111] Pélissier MA, Muller C, Hill M, Morfin R. Protection against dextran sodium
sulfate-induced colitis by dehydroepiandrosterone and 7alpha-hydroxy-
dehydroepiandrosterone in the rat. Steroids 2006;71(3):240–8.
[112] Hennebert O, Pelissier MA, Le Mée S, Wülfert E, Morfin R. Anti-inflammatory
effects and changes in prostaglandin patterns induced by 7b-hydroxy-
epiandrosterone in rats with colitis. J Steroid Biochem Mol Biol
2008;110:255–62.
[113] Davidson J, Wulfert E, Rotondo D. 7beta-hydroxy-epiandrosterone
modulation of 15-deoxy-delta12, 14-prostaglandin J2, prostaglandin D2
and prostaglandin E2 production from human mononuclear cells. J Steroid
Biochem Mol Biol 2008;112(4–5):220–7.
[114] Le Mée S, Hennebert O, Ferrec C, Wülfert E, Morfin R. 7b-Hydroxy-
epiandrosterone-mediated regulation of the prostaglandin synthesis
pathway in human peripheral blood monocytes. Steroids 2008;73:1148–59.
[115] Schmidt M, Kreutz M, Loffler G, Scholmerich J, Straub RH. Conversion of
dehydroepiandrosterone to downstream steroid hormones in macrophages. J
Endocrinol 2000;164:161–9.
[116] Dulos J, Verbraak E, Bagchus WM, Boots AM, Kaptein A. Severity of murine
collagen-induced arthritis correlates with increased CYP7B activity:
enhancement of dehydroepiandrosterone metabolism by interleukin-1beta.
Arthritis Rheum 2004;50(10):3346–53.
[117] Dulos J, van der Vleuten MA, Kavelaars A, Heijnen CJ, Boots AM. CYP7B
expression and activity in fibroblast-like synoviocytes from patients with
rheumatoid arthritis: regulation by proinflammatory cytokines. Arthritis
Rheum 2005;52(3):770–8.
[118] Dulos J, Boots AH. DHEA metabolism in arthritis: a role for the p450 enzyme
Cyp7b at the immune-endocrine crossroad. Ann NY Acad Sci
2006;1069:401–13.
[119] Hill M, Havlíková H, Vrbíková J, Kancheva R, Kancheva L, Pouzar V, Cerný I,
Stárka L. The identification and simultaneous quantification of 7-
hydroxylated metabolites of pregnenolone, dehydroepiandrosterone, 3beta,
17betaandrostenediol, and testosterone in human serum using gas
chromatography-mass spectrometry. J Steroid Biochem Mol Biol
2005;96:187–200.
[120] Offner H, Zamora A, Drought H, Matejuk A, Auci DL, Morgan EE, Vandenbark
AA, Reading CL. A synthetic androstene derivative and a natural androstene
metabolite inhibit relapsing-remitting EAE. J Neuroimmunol
2002;130:128–39.
[121] Auci DL, Ahlem C, Li M, Trauger R, Dowding C, Paillard F, Frincke J, Reading CL.
The immunobiology and therapeutic potential of androstene hormones and
their synthetic derivatives: novel anti-inflammatory and immune regulating
steroid hormones. Mod Asp Immunobiol 2003;3:64–70.
[122] Marcu AC, Kielar ND, Paccione KE, Barbee RW, Carter H, Ivatury RR,
Diegelmann RF, Ward KR, Loria RM. Androstenetriol improves survival in a
rodent model of traumatic shock. Resuscitation 2006;71:379–86.
[123] Offner H, Firestein GS, Boyle DL, Pieters R, Frincke JM, Garsd A, White SK,
Reading CL, Auci DL. An orally bioavailable synthetic analog of an active
dehydroepiandrosterone metabolite reduces established disease in rodent
models of rheumatoid arthritis. J Pharmacol Exp Ther 2009;329(3):1100–9.
[124] Schmidt M, Naumann H, Weidler C, Schellenberg M, Anders S, Straub RH.
Inflammation and sex hormone metabolism. Ann NY Acad Sci
2006;1069:236–46.
[125] Auci DL, Reading CL, Frincke JM. 7-Hydroxy androstene steroids and a novel
synthetic analogue with reduced side effects as a potential agent to treat
autoimmune diseases. Autoimmun Rev 2009;8:369–72.
[126] Canning MO, Grotenhuis K, de Wit HJ, Drexhage HA. Opposing effects of
dehydroepiandrosterone and dexamethasone on the generation of
monocyte-derived dendritic cells. Eur J Endocrinol 2000;143:687–95.
 L.E. Kihel / Steroids 77 (2012) 10–26
[127] Harding G, Mak YT, Evans B, Cheung J, Macdonald D, et al. The effects of
dexamethasone and dehydroepiandrosterone (DHEA) on cytokines and
receptor expression in a human osteoblastic cell line: Potential steroid-
sparing role for DHEA. Cytokine 2006;36:57–68.
[128] Wang L, Wang YD, Wang WJ, Zhu Y, Li DJ. Dehydroepiandrosterone improves
murine osteoblast growth and bone tissue morphometry via mitogen-
activated protein kinase signaling pathway independent of either androgen
receptor or estrogen receptor. J Mol Endocrinol 2007;38:467–79.
[129] Hartkamp A, Geenen R, Godaert GL, Bijl M, Bijlsma JW, Derksen RH. The effect
of dehydroepiandrosterone on lumbar spine bone mineral density in patients
with quiescent systemic lupus erythematosus. Arthritis Rheum
2004;50(11):3591–5.
[130] Sánchez-Guerrero J, Fragoso-Loyo HE, Neuwelt CM, Wallace DJ, Ginzler EM,
Sherrer YR, et al. Effects of prasterone on bone mineral density in women
with active systemic lupus erythematosus receiving chronic glucocorticoid
therapy. J Rheumatol 2008;35(8):1567–75.
[131] Von Mühlen D, Laughlin GA, Kritz-Silverstein D, Bergstrom J, Bettencourt R.
Effect of dehydroepiandrosterone supplementation on bone mineral density,
bone markers, and body composition in older adults: the DAWN trial.
Osteoporos Int 2008;19(5):699–707.
[132] Urban NH, Chamberlin B, Ramage S, Roberts Z, Loria RM, Beckman MJ. Effects
of a/b-androstenediol immune regulating hormones on bone remodeling and
apoptosis in osteoblasts. J Steroid Biochem Mol Biol 2008;110(3–5):223–9.
[133] Boyce BF, Xing L. Biology of RANK, RANKL, and osteoprotegerin. Arthritis Res
Ther 2007;9(Suppl 1):S1.
[134] Malik AK, Khaldoyanidi S, Auci DL, Miller SC, Ahlem CN, Reading CL, Page T,
Frincke JM. 5-Androstene-3b, 7b, 17b-triol (b-AET) slows thermal injury
induced osteopenia in mice. relation to aging and osteoporosis. PLoS One
2010;5(10):e13566.
[135] Su CY, Lardy H. Induction of hepatic mitochondrial glycerophosphate
dehydrogenase in rats by dehydroepiandrosterone. J Biochem
1991;110(2):207–13.
[136] Bobyleva V, Kneer N, Bellei M, Batelli D, Lardy H. Concerning the mechanism
of increased thermogenesis in rats treated with dehydroepiandrosterone. J
Bionerg Biomembr 1993;25(3):313–21.
[137] Marwah P, Marwah A, Kneer N, Lardy H. Ergosteroids IV: synthesis and
biological activity of steroid glucuronosides, ethers, and alkylcarbonates.
Steroids 2001;66(7):581–95.
[138] Ihler G, Chami-Stemman H. 7-Oxo-DHEA and Raynaud’s phenomenon. Med
Hypotheses 2003;60:391–7.
[139] Lardy H, Partridge B, Kneer N, Wei Y. Ergosteroids: induction of thermogenic
enzymes in liver of rats treated with steroids derived from
dehydroepiandrosterone. Proc Natl Acad Sci USA 1995;92(14):6617–9.
[140] Bobyleva V, Bellei M, Kneer N, Lardy H. The effects of the ergosteroid 7-oxo-
dehydroepiandrosterone on mitochondrial membrane potential: possible
relationship to thermogenesis. Arch Biochem Biophys 1997;341(1):122–8.
[141] Tagawa N, Takano T, Fukata S, Kuma K, Tada H, Izumi Y, et al. Serum
concentrations of androstenediol and androstenediol sulfate in patients with
hyperthyroidism and hypothyroidism. Endocr J 2001;48(3):345–54.
[142] Sulcová J, Hampl R, Hill M, Stárka L, Novácek A. Delayed effects of short-term
transdermal application of 7-oxo-dehydroepiandrosterone on its
metabolites, some hormonal steroids and relevant proteohormones in
healthy male volunteers. Clin Chem Lab Med 2005;43(2):221–7.
[143] Hampl R, Sulcová J, Bílek R, Hill M. How short-term transdermal treatment of
men with 7-oxo-dehydroepiandrosterone influence thyroid function. Physiol
Res 2006;55(1):49–54.
[144] Drbalová K, Matucha P, Matejková-Behanová M, Bílek R, Kríz L, Kazihnitková
H, Hampl R. Immunoprotective steroids and SHBG in non-treated
hypothyroidism and their relationship to autoimmune thyroid disorders.
Physiol Res 2008;57:S119–25.
[145] Ntambi JM, Kim YC. Adipocyte differentiation and gene expression. J Nutr
2000;130(12):3122S–6S.
[146] Rice SP, Zhang L, Grennan-Jones F, Agarwal N, Lewis MD, Rees DA, Ludgate M.
Dehydroepiandrosterone (DHEA) treatment in vitro inhibits adipogenesis in
human omental but not subcutaneous adipose tissue. Mol Cell Endocrinol
2010;320(1–2):51–7.
[147] Gomez FE, Miyazaki M, Kim YC, Marwah P, Lardy HA, Ntambi JM, Fox BG.
Molecular differences caused by differentiation of 3T3–L1 preadipocytes in
the presence of either dehydroepiandrosterone (DHEA) or 7-oxo-DHEA.
Biochemistry 2002;41(17):5473–82.
[148] Marwah A, Gomez FE, Marwah P, Ntambi JM, Fox BG, Lardy H. Redox
reactions of dehydroepiandrosterone and its metabolites in differentiating
3T3–L1 adipocytes: A liquid chromatographic–mass spectrometric study.
Arch Biochem Biophys 2006;456(1):1–7. Erratum in: Arch Biochem Biophys
2007; 465(1):302.
[149] Chen W, Thiboutot D, Zouboulis CC. Cutaneous androgen metabolism: basic
research and clinical perspectives. J Invest Dermatol 2002;119:992–1007.
[150] Labrie F, Cusan L, Gomez JL, Martel C, Bérubé R, Bélanger P, et al. Changes in
serum DHEA and eleven of its metabolites during 12-month percutaneous
administration of DHEA. J Steroid Biochem Mol Biol 2008;110(1–2):1–9.
[151] Labrie, Luu-the V, Labrie C, Pelletier G, El-Alfy M. Intracrinology and the skin.
Horm Res 2000;54:218–29.
[152] Lee KS, Oh KY, Kim BC. Effects of dehydroepiandrosterone on collagen and
collagenase gene expression by skin fibroblasts in culture. J Dermatol Sci
2000;23:103–10.
25
[153] Shin MH, Rhie GE, Park CH, Kim KH, Cho KH, Eun HC, et al. Modulation of
collagen metabolism by the topical application of dehydroepiandrosterone to
human skin. J Invest Dermatol 2005;124:315–23.
[154] Calvo E, Luu-The V, Morissette J, Martel C, Labrie C, Bernard B, Bernerd F,
Deloche C, Chaussade V, Leclaire J, Labrie F. Pangenomic changes induced by
DHEA in the skin of postmenopausal women. J Steroid Biochem Mol Biol
2008;112(4–5):186–93.
[155] Luu-the V, Ferraris C, Duche D, Bélanger P, Leclaire J, Labrie F. Steroid
metabolism and profile of steroidogenic gene expression in Episkin: high
similarity with human epidermis. J Steroid Biochem Mol Biol 2007;107(1–
2):30–6.
[156] Mizushima Y, Wang P, Jarrar D, Cioffi WG, Bland KI, Chaudry IH. Estradiol
administration after trauma-hemorrhage improves cardiovascular and
hepatocellular functions in male animals. Ann Surg 2000;232:673–9.
[157] Ba ZF, Wang P, Koo DJ, Cioffi WG, Bland KI, Chaudry IH. Alterations in tissue
oxygen consumption and extraction after trauma and hemorrhagic shock.
Crit Care Med 2000;28:2837–42.
[158] Ba ZF, Kuebler JF, Rue III LW, Bland KI, Wang P, Chaudry IH. Gender dimorphic
tissue perfusion response after acute hemorrhage and resuscitation: role of
vascular endothelial cell function. Am J Physiol Heart Circ Physiol
2003;284:H2162–9.
[159] Shimizu T, Choudhry MA, Szalay L, Rue III LW, Bland KI, Chaudry IH. Salutary
effects of androstenediol on cardiac function and splanchnic perfusion after
trauma-hemorrhage. Am J Physiol Regul Integr Comp Physiol
2004;287(2):R386–90.
[160] Shimizu T, Szalay L, Choudhry MA, Schwacha MG, Rue III LW, Bland KI,
Chaudry IH. Mechanism of salutary effects of androstenediol on hepatic
function after trauma-hemorrhage: role of endothelial and inducible nitric
oxide synthase. Am J Physiol Gastrointest Liver Physiol
2005;288(2):G244–50.
[161] Nilsson S, Mäkelä S, Treuter E, Tujague M, Thomsen J, Andersson G, et al.
Mechanisms of estrogen action. Physiol Rev 2001;81:1535–65.
[162] Matthews J, Gustafsson JÅ. Estrogen signaling: a subtle balance between ERa
and ERb. Mol Interv 2003;3:281–92.
[163] Maggiolini M, Donze O, Jeannin E, Ando S, Picard D. Adrenal androgens
stimulate the proliferation of breast cancer cells as direct activators of
estrogen receptor alpha. Cancer Res 1999;59:4864–9.
[164] Chen F, Knecht K, Birzin E, Fisher J, Wilkinson H, Mojena M, et al. Direct
agonist/antagonist functions of dehydroepiandrosterone (DHEA).
Endocrinology 2005;146:4568–96.
[165] Weihua Z, Lathe R, Warner M, Gustafsson JÅ. An endocrine pathway in the
prostate, ERb, AR, 5a-androstane-3b, 17b-diol, and CYP7B1, regulates
prostate growth. Proc Natl Acad Sci USA 2002;99:13589–94.
[166] Martin C, Ross M, Chapman KE, Andrew R, Bollina P, Seckl JR, Habib FK. CYP7B
generates a selective estrogen receptor b agonist in human prostate. J Clin
Endocrinol Metab 2004;89:2928–35.
[167] Umetani M, Domoto H, Gormley AK, Yuhanna IS, Cummins CL, Javitt NB, et al.
27-Hydroxycholesterol is an endogenous SERM that inhibits the
cardiovascular effects of estrogen. Nat Med 2007;13:1185–92.
[168] DuSell CD, Umetani M, Shaul PW, Mangelsdorf DJ, McDonnell DP. 27-
Hydroxycholesterol is an endogenous selective estrogen receptor modulator.
Mol Endocrinol 2008;22:65–77.
[169] Pettersson H, Lundqvist J, Norlin M. Effects of CYP7B1-mediated catalysis on
estrogen receptor activation. Biochim Biophys Acta 2010;1801(9):1090–7.
[170] Negro-Vilar A. Selective androgen receptor modulators (SARMs): a novel
approach to androgen therapy for the new millennium. J Clin Endocrinol
Metab 1999;84:3459–62.
[171] Miyamoto H, Yeh S, Lardy H, Messing E, Chang C. D5-Androstenediol is a
natural hormone with androgenic activity in human prostate cancer cells.
Proc Natl Acad Sci USA 1998;95(19):11083–8.
[172] Chang HC, Miyamoto H, Marwah P, Lardy H, Yeh S, Huang KE, Chang C.
Suppression of Delta(5)-androstenediol-induced androgen receptor
transactivation by selective steroids in human prostate cancer cells. Proc
Natl Acad Sci USA 1999;96(20):11173–7.
[173] Marwah P, Marwah A, Lardy HA, Miyamoto H, Chang C. C19-Steroids as
androgen receptor modulators: Design, discovery, and structure-activity
relationship of new steroidal androgen receptor antagonists. Bioorg Med
Chem 2006;14(17):5933–47.
[174] Mo Q, Lu SF, Simon NG. Dehydroepiandrosterone and its metabolites:
differential effects on androgen receptor trafficking and transcriptional
activity. J Steroid Biochem Mol Biol 2006;99(1):50–8.
[175] Pelletier G. Expression of steroidogenic enzymes and sex-steroid receptors in
human prostate. Best Pract Res Clin Endocrinol Metab 2008;22:223–8.
[176] Luu-the V, Bélanger A, Labrie F. Androgen biosynthetic pathways in the
human prostate. Best Pract Res Clin Endocrinol Metab 2008;22(2):207–21.
[177] Leskelä S, Honrado E, Montero-Conde C, Landa I, Cascón A, Letón R, et al.
Cytochrome P450 3A5 is highly expressed in normal prostate cells but absent
in prostate cancer. Endocr Relat Cancer 2007;14(3):645–54.
[178] Mitamura K, Nakagawa T, Shimada K, Namiki M, Koh E, Mizokami A, et al.
Identification of dehydroepiandrosterone metabolites formed from human
prostate homogenate using liquid chromatography–mass spectrometry and
gas chromatography–mass spectrometry. J Chromatogr A 2002; 961:97–105.
[179] Mohler JL, Gregory CW, Ford OH, Kim D, Weaver CM, Petrusz P, et al. The
androgen axis in recurrent prostate cancer. Clin Cancer Res
2004;10(1):440–8.
26 L.E. Kihel / Steroids 77 (2012) 10–26
[180] Arai S, Miyashiro Y, Shibata Y, Tomaru Y, Kobayashi M, Honma S, Suzuki K.
Effect of castration monotherapy on the levels of adrenal androgens in
cancerous prostatic tissues. Steroids 2011;76(3):301–8.
[181] Widstrom RL, Dillon JS. Is there a receptor for dehydroepiandrosterone or
dehydroepiandrosterone sulfate? Semin Reprod Med 2004;22:289–98.
[182] Martin C, Ross M, Chapman KE, Andrew R, Bollina P, Seckl JR, Habib FK. CYP7B
generates a selective estrogen receptor beta agonist in human prostate. J Clin
Endocrinol Metab 2004;89:2928–35.
[183] McPherson SJ, Ellem SJ, Simpson ER, Patchev V, Fritzemeier KH, Risbridger GP.
Essential role for estrogen receptor beta in stromal-epithelial regulation of
prostatic hyperplasia. Endocrinology 2007;148:566–74.
[184] McPherson SJ, Ellem SJ, Risbridger GP. Estrogen-regulated development and
differentiation of the prostate. Differentiation 2008;76:660–70.
[185] Risbridger GP, Ellem SJ, McPherson SJ. Estrogen action on the prostate gland:
a critical mix of endocrine and paracrine signaling. J Mol Endocrinol
2007;39:183–8.
[186] Carruba G. Estrogen and prostate cancer: an eclipsed truth in an androgen-
dominated scenario. J Cell Biochem 2007;102:899–911.
[187] Hsing AW, Chu LW, Stanczyk FZ. Androgen and prostate cancer: is the
hypothesis dead? Cancer Epidemiol Biomarkers Prev 2008;17:2525–30.
[188] Arnold JT, Blackman MR. Does DHEA exert direct effects on androgen and
estrogen receptors, and does it promote or prevent prostate cancer?
Endocrinology 2005;146:4565–7.
[189] Arnold JT. DHEA metabolism in prostate: For better or worse? Mol Cell
Endocrinol 2009;301(1–2):83–8.