ADRENAL HORMONES

SITI ANNISA DEVI TRUSDA

BIOCHEMISTRY DEPARTMENT
UNIVERSITAS ISLAM BANDUNG
Adrenal Steroid Hormones

Review of the Adrenal Gland

Review of Adrenal Steroidogenesis
Production and Roles of Cortisol
Production and Roles of Aldosterone
Adrenal Androgens
 Adrenal Steroids: A Review

• The adrenal glands are located immediately superior

to the kidneys.

• There are three classes of adrenal

steroids:
- mineralocorticoids,
- glucocorticoids, and
- androgens
 Organization of the Adrenal Gland

There is an adrenal cortex and adrenal medulla.

Steroids are made in the 3 zones of the adrenal

cortex:
mineralocorticoids: zona glomerulosa
glucocorticoids: zona fasciculata
androgens: zona reticularis
 Adrenal Steroidogenesis
• The first step is the conversion of cholesterol to
pregnenolone, which occurs in the mitochondria.
• This reaction is carried out by the enzyme,
cytochrome P450 side chain cleavage (P450scc).
This is a rate limiting, nonreversible enzymatic
step in the initiation of steroid biosynthesis.
 Adrenal Steroidogenesis
• Next, pregnenolone can be converted (in smooth ER) into
three different pathways, depending upon whether you
want to make mineralcorticoids, glucocorticoids, or
androgens:

17hydroxylase lyase
pregnenolone 17-hydroxypregnenolone dehydroepiandrosterone
-hydroxysteroid dehydrogenase
progesterone androstenedione
21-hydroxylase

11 -hydroxylase

18 hydroxylase/oxidase glucocorticoids
mineralocorticoids (cortisol)
(aldosterone)
 Adrenal Steroidogenesis
• What determines which pathway is taken?
• Each step of the pathway is regulated by a specific
enzyme.
• Different zones of the adrenal cortex have different
relative activities of enzymes, resulting in different
chemical reactions taking place.
• These enzymes are located in the smooth ER.
 Regulation of Cortisol Release
• Cortisol release is primarily under neuroendocrine control.

hypothalamus

CRF
(-)
pituitary

ACTH

cortisol adrenal gland

Some Additional Information:
 CRF acts through a Gs-protein coupled
receptor, increasing cAMP levels in
pituitary corticotroph cells.
 Recall that ACTH is produced by post-
translational modification of
proopiomelanocortin:
MSH MSH clip LPH Endorphin
}
ACTH
 Action of ACTH on Cortisol Production

• ACTH binds to the ACTH receptor (Gs-coupled), resulting in

increased cyclic AMP and activation of PKA pathway.
• This results in:
- increased conversion of cholesterol esters to free
cholesterol
- increased expression of steroidogenic acute regulatory
protein (cholesterol transfer)
- increased expression of P450 side chain cleavage
- increased production of IGF-II (stimulation of cell growth)
Action of ACTH on Cortisol Production
ACTH

AC
Gs growth
ATP cyclic AMP

cholesterol
ester PKA
IGF-II
P450scc
pregnenolone
free cholesterol

cortisol
 Daily Pattern of Cortisol Release
• Changes in cortisol levels occur about 30 minutes
after corresponding changes in ACTH.
• Cortisol usually increases a few hours after sleep,
then declines (it has a diurnal rhythm).
• Also an increase shortly after wakening in the
morning, and sporadically throughout the day.
• Peaks shortly after we rise in morning.
• Lowest shortly before and shortly after sleep
begins.
• Circadian rhythm may reflect actions required for
the stress of simply living.
Cortisol Effects: Body Responses to the Stress of Living
Cortisol Circadian Rhythm
 Transport of Cortisol

• Recall that the majority of cortisol is bound to

cortisol binding globulin (also called transcortin).
Only 5% of circulating cortisol is in free form.
• Also recall that cortisol bound to CBG is not
biologically available.
• CBG is produced by the liver.
• Production of CBG is increased by estradiol.
 Metabolism of Cortisol
• Cortisol has a half-life of about 90 minutes.
• Cortisol is metabolized in the liver, where it is
conjugated to a glucuronic acid.
- conjugation increases solubility in water
- conjugated hormone is excreted via the
kidneys
 Functions of Cortisol

• Cortisol is the primary glucocorticoid in humans.

• Cortisol is essential for life.
• Roles of cortisol:
- carbohydrate metabolism
- permissive effects on body functions and
development (epinephrine release, lipid
metabolism)
- mineralocorticoid activity at the kidney
(minor)
- anti-inflammatory compound
 Control of Cortisol Secretion: Feedback Loops

• External stimuli
• Hypothalamic
• Anterior Pituitary
• Adrenal cortex
• Tissues

Figure 23-3: The control pathway for cortisol

 Role in Carbohydrate Metabolism
• Cortisol has effects on nutrient metabolism:
- decreases uptake of circulating glucose by muscle and
adipose tissue
- proteolysis of muscle protein for energy
- increases lipolysis in adipose tissue for energy
- at the liver, get gluconeogenesis from free fatty acids
and amino acids produced by actions in muscle and
adipose tissue (increases PEPCK activity)

Overall, increases plasma glucose levels.

 Role in Carbohydrate Metabolism

• Cortisol release is inhibited by high blood

glucose.
 Permissive Actions of Cortisol

• Cortisol is required for:

- synthesis of epinephrine (adrenal medulla)
- normal vasoconstriction (absence leads to
decreased blood pressure)
- normal glomerular filtration rate at kidney
(related to blood pressure)
 Effects of Elevated Cortisol on Bone

• Recall that high glucocorticoid levels cause

decreased bone mass. This is due to:
– antagonizing the effects of vitamin D on
calcium uptake
– inhibiting collagen synthesis
– synergizing with PTH to break down bone
 Mineralocorticoid Activity of Cortisol

• Under normal conditions of blood pressure and

sodium levels, cortisol has some
mineralocorticoid activity (increasing sodium and
water reabsorption at the kidney).
• Cortisol has low affinity for the mineralocorticoid
receptor.
• However, much more cortisol than aldosterone is
secreted.
 Cortisol as an Anti-Inflammatory Agent
• The body responds to bacteria or tissue damage with an
inflammatory response:
- increased production of chemicals such as interleukins
- these cause increased vasodilation and blood flow to the
area
- increased blood flow brings in phagocytes, mast cells,
and lymphocytes
- these cells trigger lysosomal reactions, histamine
release, and collagenase production
• Result: destruction of bacteria; healing and remodeling of
tissue.
 Cortisol as an Anti-Inflammatory Agent
• The painful effects of healing/irritation can be treated
with cortisol.
• Cortisol inhibits inflammatory and immune responses:
- decreased permeability of capillaries (decreasing
entrance of cells to inflammed areas)
- stabilization of lysosomal membranes (decreased release
of lysozomes)
- decreased phagocytosis by white blood cells
- suppression of T lymphocytes (immune cells)
- decreased synthesis of interleukins
- inhibits synthesis of collagenase
• Results in slower, but less painful healing.
 Cortisol and Chronic Stress

• Prolonged exposure to high cortisol levels

can lead to break down of muscle,
excessive epinephrine release,
hyperglycemia, weakening of bone,
destruction of the immune system,
inhibition of reproductive function, and
other complications.
The General Adaptation Syndrome
The General Adaptation Syndrome
The General Adaptation Syndrome
 Permissive Effects of Cortisol on
Development
• Cortisol is required for normal development:
- permissive role in final maturation of many organs
- required for synthesis of digestive enzymes,
surfactant
- required for skeletal growth in children
 Mechanisms of Cortisol Action
• The actions of cortisol are mediated through the
glucocorticoid receptor.
• Intracellular receptor in steroid receptor superfamily.
• Stimulates transcription of target genes by interaction of
bound receptor with GRE in 5’ flanking region.
• Inhibits transcription of some genes by interaction of
receptor with AP1 (jun/fos dimer), decreasing AP1-
mediated gene expression.

AP1
transcription
cortisol GR
AP1
site
 Hormone Effects on Gene Activity

Cortisol
 Cushing’s

• Hypercortisolism
• Can be caused by pituitary or adrenal tumors
• Symptoms include central obesity but thin limbs;
rounded face; weak bones; weak muscles; high
blood pressure; high blood glucose; fragile skin;
mood swings…
 Exogenous
Iatrogenic CRH Hydrocortisone
Cushing’s Syndrome (+)

(-)

ACTH (- ) (-) Cortisol

 ACTH-dependent CRH
(+)
Cushing’s disease

Autonomous ACTH secreting tumour

ACTH (+) (-) Cortisol

 CRH
Adrenocortical tumour (+)

ACTH (-) (+) Cortisol

Autonomous cortisol secreting tumour

 CRH
Ectopic ACTH (+)
syndrome

Ectopic ACTH secreting tumour

(+) Cortisol

ACTH (+)
 CRH (+) Ectopic CRH secreting
Ectopic CRH tumour
producing tumour

ACTH (+) (+) Cortisol

DIAGNOSIS
Urinary free cortisol (UFC)

• Free cortisol may be detected by

– Structurally-based techniques (eg, high
performance liquid chromatography).
– Antibody-based techniques (immunoassays):
less specific since antibodies may cross-react
with other steroids.
• When several UFC collections are normal,
CS is unlikely.
• May use early morning UFC/creatinine
(nmol/l:mmol/l) ratio of greater than 50 is
suggestive of CS.
• Four-fold greater than the upper limit of
normal, is considered diagnostic test.
• Sensitivity of 94.4%, false negative 5.6%
and a false positive 3.3%
– 315 patients with Cushing's syndrome and 479 lean, obese, or
chronically ill patients who did not have Cushing's syndrome
Crapo L, Metabolism 1979

• Diagnostic sensitivity 100% and specificity

98%
– 48 patients with Cushing's syndrome, 95 obese, and 94 normal
subjects
Mengden Clin Invest 1992
• Sensitivity problem
• In patients with mild CS, UFC levels may be consistently normal, .

Trainer Lancet 2000

• 10% to 15% of patients with the Cushing syndrome, at least one of
four 24-hour determinations of urine free cortisol level are within the
normal range
Nieman Endocrine Soc; 1990
• Specificity problem
• Elevated UFC levels may also be found in:
– ETOH, Phenytoin, Phenobarbital, primidon
– Pregnant women
– 40-60% of depressed inpatients
– Patients with Polycystic Ovarian Syndrome (PCOS)
Carroll 1976, Cizza 1996, and Yanovski 1993
Salivary cortisol levels

• Many studies have demonstrated great

promise In the use of this test as a
screening test for CS
– More than 140 patients found an Increased
bedtime salivary cortisol levels yield both a
• Sensitivity of 93%
• Specificity of 100%
Papanicolaou J Clin Endocrinol Metab. 2002
Low-Dose Dexamethasone Suppression Test

• 1mg of dexamethasone at 23.00 hours and

measurement of plasma cortisol at 08.00 or
09.00 hours the next morning.
– High diagnostic accuracy with a sensitivity of
98% using a post-dexamethasone serum
cortisol value of less than 50nmol/l (1.8µg/l)
• Consensus opinion in the United Kingdom: value
of less than 50nmol/l (1.8µg/l) effectively
Excludes the Cushing syndrome

Wood Ann Clin Biochem1997

• False positive results can occur because:
– Failure to take dexamethasone as prescribed.
– Accelerated hepatic metabolism
• Phenytoin, Carbamazepine, Barbiturates,
Aminoglutethimide or Rifampicin), and ETOH.
– Increased concentration of cortisol binding
globulin (CBG)
• Pregnancy or Estrogen treatment.
Dexamethasone-CRH Test
• Dexamethasone (0.5 mg Q 6 hours) is given X8, the first dose at
noon and the last dose at 6:00 a.m.

• Corticotropin-releasing hormone CRH (1µg/kg) is then administered

IV at 8:00 a.m., and plasma cortisol and ACTH levels are obtained
at 15-minute intervals for 1 hour.

• Cortisol level greater than 39 nmol/L (1.4 g/dL) measured 15

minutes after the administration of CRH correctly identifies patients
with the Cushing syndrome, and levels of 39 nmol/L or less (1.4
g/dL) are considered normal.

• ??Normal ACTH response.

– Patients with the Cushing syndrome usually have a peak ACTH
response exceeding 3.3 pmol/L (15 pg/mL) during the test.
• The dexamethasone-CRH test is usually
reserved for patients with equivocal results
on other diagnostic tests and a high index
of suspicion for the Cushing syndrome.
Measurement of ACTH
• IRMA (Immunoradiometric assay) is more
sensitive and specific assay than RIA
(radioimmunoassay) for ACTH.

• Some tumors secrete active Large ACTH

fragments not detected by IRMA; therefore RIA
is preferred for initial evaluation.
Wallach, 7th edi, 2000
Measurement of ACTH
• A suppressed ACTH concentration <10pg/ml at
0900 hours, with concomitant increased cortisol
production indicates adrenal-dependent Cushing
syndrome (ACTH-Independent) caused by
classic negative feedback both at the
hypothalamus (to decrease CRH release) and at
the pituitary (to decrease ACTH release).

• Plasma ACTH levels greater than 4.4 pmol/L (20

pg/mL) imply an ACTH-dependent cause
• Values between 1.1 and 4.4 pmol/L (5 to
20 pg/mL) usually require a CRH
stimulation test.
– Patients with ACTH independent Cushing
syndrome usually have a subnormal peak
ACTH response to CRH stimulation (usually <
6.6 pmol/L [30 pg/mL]).
 Functions of Aldosterone
• Aldosterone plays a major role in mineral and water
balance:
-increases sodium reabsorption in distal tubules of
the kidney
-as sodium is actively reabsorbed, also get passive
absorption of water by osmosis
-also get excretion of potassium (active and passive)
- result: increased retention of sodium and of water

• Aldosterone also increases sodium reabsorption in

sweat glands.
Overview
 Mechanism of Aldosterone Action
• Aldosterone acts through the intracellular
mineralocorticoid receptor.
• Binding of hormone to the receptor results in
expression of gene products that influence the rate of
Na+ transport from urine back into the kidney.
• Na+ and H2O reabsorption:
If H2O reabsorption is impaired, and [Na+]
increases, the increased osmotic pressure will
activate ADH  increase renal H2O reabsorption 
increase H2O intake by stimulating thirst.
• These target genes result in:
- increased availability of ATP to drive active
transport
- increased synthesis of Na+ transporters
- increased leakiness of the membrane to Na+,
so that Na+ is reabsorbed by diffusion
 Influence of Aldosterone on Na+ and H2O
Reabsorption and K+ Excretion
Na+ Na+
Aldosterone H+
H2O H2O

K+ K+
Na+ Na+
K+ K+

epithelial cell distal tubule

 Binding of Cortisol to the Mineralocorticoid
Receptor

• In the laboratory, cortisol can bind very effectively to the

mineralocorticoid receptor, resulting in activation.
• However, in most tissues that express the
mineralocorticoid receptor, most of the cortisol is locally
converted to cortisone by 11-beta hydroxysteroid
dehydrogenase.
• Cortisone does not bind to the mineralocorticoid receptor
very well.
• Deficiency in 11-beta hydroxysteroid dehydrogenase leads
to high blood pressure, due to increased
mineralocorticoid activity of cortisol (genetic defect,
licorice).
 Role of 11-beta Hydroxysteroid
Dehydrogenase in Cortisol Metabolism

cortisol mineralocorticoid receptor

11-beta HSD

increased retention of sodium

cortisone and water from kidney
 Regulation of Aldosterone Production

• The main stimulator of aldosterone production is

angiotensin II, not ACTH.

• ACTH does have a trophic (stimulatory) effect on

the zona glomerulosa, preventing atrophy.
 The Renin-Angiotensin System

• The juxtaglomerular cells of the kidney

release renin in response to decreased
blood pressure, decreased sodium levels,
or sympathetic stimulation.
 The Renin-Angiotensin System

• Renin converts angiotensinogen (from the liver)

into angiotensin I

• Angiotensin I is converted to angiotensin II in a

number of places in the body (ie, lungs)

• Angiotensin II acts on adrenal cells to stimulate

aldosterone release, and is itself a vasoconstrictor
(increases blood pressure)
 The Renin-Angiotensin System

decreased Na+ angiotensinogen

(macula densa) (liver)
(-)
juxtaglomerular renin
cells
decreased
blood pressure angiotensin I
kidney
(-)
tubules
Na+
angiotensin II
H2O

adrenal
aldosterone
 Transport and Metabolism of Aldosterone

• There is no specific aldosterone carrier protein

• As a result, most aldosterone is biologically available, and
only low levels of aldosterone release are required each day
• The circulating half life of aldosterone is very short.

• Aldosterone is metabolized in the liver, conjugated with

glucoronide and excreted in the urine
 Role of Atrial Natriuretic Factor (ANF)
• ANF is produced by atria of the heart, and production is increased
by increased blood pressure.
• ANF inhibits aldosterone production (to inhibit resorption of water)
(blocks actions of angiotensin II).
• ANF also has these effects:
- inhibits renin release (blocks production of angiotensin II)
- decreases AVP release from posterior pituitary
- causes dilation of blood vessels to decrease bp
- acts directly on the kidney to increase loss of water and sodium
- increased glomerular filtration rate
- decreased reabsorption of sodium

Net result: decreased water volume, decreased bp.

 Adrenal Androgens
• Produced in the zona reticularis.
• Production is stimulated by ACTH.
• Main product: androstenedione
• In females, adrenal androgens may play roles in
- development of axillary and pubic hair
- libido (sexual desire)
(In men, androgens from the testis serve these functions)
• Adrenal androgens may also be peripherally converted
(in adipose tissue, for example) into testosterone and
estrone.
 Conn’s Disease
• Primary hyperaldosteronism
• Caused by adrenal tumor
• Main symptom: hypertension (up to 15% of
clinical hypertension cases?)
 Congenital Adrenal Hyperplasia
- Deficiency in 3-HSD (type II), 11-hydroxylase, or
21-hydroxylase activity blocks cortisol formation,
leading to increased ACTH (less negative feedback)
- Result: Increased formation of androstenedione or
DHEA (which is converted to androstenedione by 3-
HSD type I in liver)
17hydroxylase lyase
pregnenolone 17-hydroxypregnenolone dehydroepiandrosterone
-hydroxysteroid dehydrogenase
progesterone androstenedione
21-hydroxylase

11 -hydroxylase

18 hydroxylase/oxidase glucocorticoids
mineralocorticoids (cortisol)
(aldosterone)
ALHAMDULILLAH

HATURNUHUN