Molecular and Cellular Endocrinology 304 (2009) 30–42

Contents lists available at ScienceDirect

Molecular and Cellular Endocrinology

journal homepage: www.elsevier.com/locate/mce

Review

Soy, phytoestrogens and metabolism: A review

Christopher R. Cederroth, Serge Nef ∗
Department of Genetic Medicine and Development, University of Geneva Medical School, 1211 Geneva 4, Switzerland

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

Article history: Of any plant, soy contains the largest concentration of isoflavones, a class of phytoestrogens. Phytoestro-
Received 14 January 2009 gens are structurally similar to estradiol and mimic its effects. Soy and phytoestrogens receive increasing
Accepted 24 February 2009 attention due to the health benefits associated with their consumption. Here we review the data collected
on the effects of soy and phytoestrogens on glucose and lipid metabolism and their possible mechanisms
Keywords: of action. Overall, there is a suggestive body of evidence that soy and dietary phytoestrogens favorably
Dietary soy
alter glycemic control, improve weight and fat loss, lower triglycerides, low density lipoprotein (LDL)
Phytoestrogens
cholesterol and total cholesterol. However, these results must be interpreted with care, and additional
Isoflavones
Genistein
evidence is needed before a firm conclusion can be drawn. In particular, since not all activities related
Endocrine disruptor to soy can be assigned to the estrogenic-like activity, further studies are needed to identify firstly which
Obesity soy constituent(s) improve metabolic parameters when ingested and secondly, which are the mecha-
Glycemia nisms whereby dietary soy improves metabolic-related conditions like obesity and diabetes. Finally, the
potential detrimental effects of soy and phytoestrogens are briefly discussed.
© 2009 Elsevier Ireland Ltd. All rights reserved.

Contents

1. Metabolic diseases and therapeutic alternatives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

2. Soybean composition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
3. Absorption and metabolism of isoflavones . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
4. Soy consumption and phytoestrogen levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
5. Phytoestrogens: complex hormetic compounds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
6. Role of estrogens in metabolism. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
7. Effects of soy protein and phytoestrogens on human metabolism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
8. Actions of soy on metabolism in rodents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
9. Central actions of phytoestrogens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
10. Potentially adverse effects in consuming soy and soy-derived phytoestrogens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
11. Summary and conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

1. Metabolic diseases and therapeutic alternatives to surpass malnutrition and infectious diseases as the most sig-
nificant contributor to worldwide morbidity. In western societies,
Obesity and its related disorders, such as type 2 diabetes, car- for the first time in modern history, life expectancy of newborns is
diovascular diseases (CVD), high blood pressure, dyslipidemia [high declining as a result of these metabolic disorders (Olshansky et al.,
levels of circulating triacylglycerols and low density lipoprotein 2005). The rapid increase in obesity suggests that life-style factors
(LDL) cholesterol, and low levels of high density lipoprotein (HDL) such as high-calorie diets, physical inactivity and potentially envi-
cholesterol], have recently become a major health problem reaching ronmental endocrine disruption, rather than genetics, are the most
pandemic proportions (Engelgau et al., 2004). These diseases, com- plausible causes.
monly referred to as the Metabolic Syndrome (MS), are beginning Although the source of metabolic disorders is often the diet
itself, nutrition can also form part of the solution, in fact provid-
ing health benefits. Usually dietary intervention to control excess
∗ Corresponding author. body weight, hyperglycemia and dyslipidemia has included low
E-mail address: Serge.Nef@medecine.unige.ch (S. Nef). energy and low fat diets, but these are of limited efficacy due to

0303-7207/$ – see front matter © 2009 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.mce.2009.02.027
 C.R. Cederroth, S. Nef / Molecular and Cellular Endocrinology 304 (2009) 30–42
the strict and long-term commitment required. However, long term
health benefits can be gained from dietary proteins and bioac-
tive non-nutrients, called phytochemicals, which could be either
incorporated into the diet or be part of the food itself. These phyto-
chemicals are biologically active plant-derived compounds, which
structurally and functionally mimic estrogens (Dixon, 2004). Phy-
toestrogens are found in numerous fruits and vegetables and are
categorized into three classes, namely the isoflavones, lignans and
coumestans. While phytoestrogens are ubiquitous within the plant
kingdom, isoflavones are mainly found in the soybean—the most
important dietary source of phytoestrogens for humans, cattle and
rodents. Isoflavones have a non-steroidal structure but possess a
phenolic ring that enables them to bind the estrogen receptor (ER)
and act either as estrogen agonists or antagonists (Makela et al.,
1994, 1995).
The fact that isoflavones have been shown to exert estrogenic
effects raises the possibility that this class of phytochemicals
may affect glucose and lipid metabolism. In fact, estradiol itself
is a well known modulator of glucose homeostasis, which also
affects obesity development. For instance, postmenopausal women
develop visceral obesity and insulin resistance and are at an
increased risk of diabetes but estrogen replacement therapy nor-
malizes these abnormalities (Ahmed-Sorour and Bailey, 1980;
Bailey and Ahmed-Sorour, 1980; Gambacciani et al., 1997). From
genetic studies in rodents, it has been shown that these effects
are mediated by estrogen receptors (see below). This has caused
researchers to focus on the identification of Selective Estro-
gen Receptor Modulators (SERMs) that could be of potential
therapeutic interest for the treatment of metabolic disorders,
without having negative effects. Studies in humans and rodents
support the hypothesis that soy proteins or soy-derived phytoe-
strogens may be beneficial for the prevention of obesity and
diabetes (Bhathena and Velasquez, 2002; Velasquez and Bhathena,
2007).
The complex interactions between soy proteins and isoflavones
are fairly well understood. To understand these intricate relation-
ships, one must assess the biological activity of soy components,
both in isolation and in combination. So far, few studies have
shown that pure soy-proteins or soy proteins isolate (SPI) alone
(in absence of isoflavones) can provide beneficial metabolic effects
(Velasquez and Bhathena, 2007). The majority of the studies using
SPI remain difficult to interpret because of the lack of clarity con-
cerning the presence or absence of isoflavones in the diet. On
the other hand, soy-derived phytoestrogens have received more
attention mainly due to their benefits in decreasing age related
diseases (e.g. osteoporosis, cardiovascular disease), or hormono-
dependent cancers (e.g. prostate) (Setchell, 1998; Tham et al.,
1998; Sacks et al., 2006). Concerning metabolism, the American
Food and Drug Administration (FDA) authorized in 1999 the label-
ing of health claims on food containing soy proteins, referring to
the beneficial role of soy protein in reducing the risk of coro-
nary heart disease (CHD). The beneficial effects on metabolism
in humans have been hotly debated, but studies in rodents may
help in identifying the biologically relevant soy components and
the intimate mechanisms involved. The purpose of this review is
to examine the evidence regarding the use of soy and phytoestro-
gens in the prevention of obesity and diabetes mellitus in animals
and humans. We also discuss the mechanisms by which soy and
dietary phytoestrogens may affect glucose and lipid metabolism
and improve the control of body weight and glucose homeosta-
sis. To provide context and the requisite background information,
we begin with a brief overview about soybeans, the nutritional
composition of soy. We also present scientific evidence both in
humans and rodents supporting or refuting the potential ben-
eficial effects of soy and phytoestrogens on glucose and lipid
metabolism.
31
2. Soybean composition
Soybean (Glycine max) is composed of macronutriments such
as lipids, carbohydrates and proteins. Soybean lipids, which are
deprived of cholesterol, contain about 15% of saturated fat, 61% of
polyunsaturated fat, and 24% of monounsaturated fat (USDA, 1979).
Carbohydrates make up about 30% of the seed, with 15% being solu-
ble carbohydrates (sucrose, raffinose, stachyose) and 15% insoluble
carbohydrates (dietary fiber). The protein content of soybean varies
from 36% to 46% depending on the variety (Garcia et al., 1997;
Grieshop and Fahey, 2001; Grieshop et al., 2003). Storage proteins
are predominant, such as the 7S globulin (␤-conglycinin) and 11S
globulin (glycinin), which represent about 80% of total protein con-
tent, as well as less abundant storage proteins such as 2S, 9S, and
15S globulins (Garcia et al., 1997). Interestingly, ␤-conglycinin but
not glycinin is capable of improving serum lipid profiles in mice and
humans, in the absence of phytoestrogens (Moriyama et al., 2004;
Kohno et al., 2006).
Soybean also contains micronutriments, which include
isoflavones, phytate, soyaponins, phytosterol, vitamins and
minerals. Although beneficial effects of micronutriments such as
saponins and phytosterols on cholesterol levels and absorption
have been reported (Oakenfull, 2001; Lukaczer et al., 2006), there
is an increasing body of literature suggesting that isoflavones may
additionally have a beneficial role in lipid and glucose metabolism.
Soybeans are the most abundant source of isoflavones in food.
Studies have shown that there is a large variability in isoflavone
content and composition in soybeans. This is function of the
variety of soy grown, as well as environmental conditions (Wang
and Murphy, 1994; Caldwell et al., 2005). Abiotic and biotic stresses
such as variation in temperature, drought or nutritional status,
pest attack or light conditions may modify isoflavone content
and composition. As a consequence, total isoflavone content may
vary up to 3-fold with growth of the same soy cultivar in different
geographical areas and years (Wang and Murphy, 1994).
3. Absorption and metabolism of isoflavones
The metabolism of isoflavones is rather complex. The two major
isoflavones, genistein and daidzein, are present in soy as ␤-D-
glycosides, namely genistin and daizin (Fig. 1). These glycoside
forms are biologically inactive (Setchell, 1998). Once ingested,
isoflavone glycosides are hydrolyzed by bacterial ␤-glucosidases
in the intestinal wall, resulting in the conversion to their corre-
sponding bioactive aglycones (genistein and daidzein). Only the
aglycone forms are absorbed by the intestinal tract and are there-
fore biologically active. Daidzein can be further metabolized to
equol and O-demethyangolensin, and genistein to p-ethyl phenol.
In fact, genistein, daidzein, equol and O-demethyangolensin are the
major isoflavones detected in the blood and urine of humans and
animals (Setchell, 1998). In rodents, equol is the major circulat-
ing metabolite among isoflavones representing up to 70–90% of
all circulating isoflavones. While all rodents are equol producers,
only 30% of humans are able to metabolize daidzein into equol
(Atkinson et al., 2005). It remains unclear whether the effectiveness
of dietary phytoestrogens in reducing the risks of obesity, diabetes,
and cardiovascular disease in humans correlates with the ability of
individuals to metabolize daidzein into equol. However, it is a likely
source of variability and therefore should be taken into account
when performing clinical trials with soy or dietary phytoestrogens.
4. Soy consumption and phytoestrogen levels
In soybean, isoflavones are tightly associated with proteins.
As mentioned, the abundance of isoflavones varies according
32 C.R. Cederroth, S. Nef / Molecular and Cellular Endocrinology 304 (2009) 30–42
Fig. 1. The molecular structure of isoflavones resembles that of 17␤-estradiol. Isoflavones are found in vegetables and fruits in a biologically inactive glycoside form (genistin,
daidzin and glycitin). After ingestion, ␤-glucosidases from the intestine cleave the glucosyl residue and generate biologically active aglycones (genistein, daidzein and glycitein).
Daidzein can be further metabolized into equol.
to soy variety and culture conditions, but is also dependent
on the way soybeans have been processed. Indeed, isoflavones
can be dissociated from soy-proteins using alcohol extraction
which significantly diminishes the amount of bound-isoflavones
(Bhathena and Velasquez, 2002). This explains the substantial vari-
ability of phytoestrogen content found in soy products [0.1–5 mg
isoflavones/g of soy protein in mature and roasted soybeans,
0.3 mg/g soy protein in green soybeans and tempeh, 0.1–2 mg/g
soy protein in tofu and some soy milk preparations (Bhathena and
Velasquez, 2002)].
Numerous studies have included the investigation of the plasma
concentration of phytoestrogens and their metabolites in humans
and animals consuming a diet with or without soy (Adlercreutz
et al., 1993a; Morton et al., 1994; Coward et al., 1996). In humans
consuming soy-free diets, plasma concentration of isoflavones is
usually in the nanomolar range ≤40 nM (Morton et al., 1994; van
Erp-Baart et al., 2003). In contrast, acute ingestion of dietary soy
leads to a rapid increase in the plasma concentration of isoflavones
up to the micromolar range (Adlercreutz et al., 1993b; Xu et al.,
1994; King and Bursill, 1998; Watanabe et al., 1998). Pharmacoki-
netic studies confirm that healthy adults absorb isoflavones rapidly
and efficiently (Setchell et al., 2001). The fates of daidzein, genis-
tein and their respective ␤-glycosides are similar. The average time
taken after ingesting the aglycones to reach peak plasma concentra-
tions is 4–7 h, which is delayed to 8–11 hours for the corresponding
␤-glycosides. This suggests that the rate-limiting step for absorp-
tion is the initial hydrolysis of the glycosidic moiety. The half-life
for daidzein and genistein was reported to be 9.3 and 7.1 h respec-
tively, indicating that isoflavones or their metabolites are rapidly
excreted. Finally, factors that might influence isoflavone bioavail-
ability include intestinal microflora, food matrix, the administered
dose, intestinal transit time and the chemical composition of the
dietary isoflavones.
5. Phytoestrogens: complex hormetic compounds
In plants, the synthesis of phytoestrogens, such as soy
isoflavones, generally coincides with environmental stresses such
as pest infection, drought or lack of nutrients (Howitz and Sinclair,
2008). Recently it has been suggested that stress-induced plant
compounds upregulate stress resistance pathways in animals. This
phenomenon, called xenohormesis, proposes that chemical clues
from autotrophs (e.g. plants) provide an advance warning about
the deterioration of the environment, allowing heterotrophs (e.g.
mammals) to mount a preemptive defense response while condi-
tions are still favorable (Howitz and Sinclair, 2008). This theory has
been recently adopted to explain the health benefits provided by
stress-induced phytochemicals such as polyphenols, a group which
includes stilbenes (e.g. resveratrol found in red wine and peanuts),
catechins (e.g. epigallocatechin-3-gallate or EGCG found in green
tea), anthocyanidins, and most relevant to this review, isoflavones.
Similarly, we believe that the isoflavones genistein and daidzein
mediate most of their biological effects through the modulation of
key mammalian enzymes and receptors of stress-response path-
ways or estrogen-dependent pathways, rather than through their
well known antioxidant (Vedavanam et al., 1999) or tyrosine kinase
inhibitory properties (Akiyama et al., 1987). The affinity of phy-
toestrogens for estrogen receptors results in effects on a large
number of estrogen-regulated systems, including the cardiovascu-
lar, metabolic, reproductive, skeletal and central nervous systems.
A significant characteristic of isoflavones is their capacity to bind
both to estrogen receptors (ERs) ␣ and ␤ (Kuiper et al., 1997, 1998),
 C.R. Cederroth, S. Nef / Molecular and Cellular Endocrinology 304 (2009) 30–42 33

Table 1
Relative binding affinity (RBA) and relative transactivation activity (RTA) of various estrogenic compounds, isoflavones and endocrine disruptors in comparison with 17␤-
estradiol.

Compound Relative binding affinity Relative transactivation

ER␣ ER␤ ER␣ ER␤

17␤-estradiol 100 100 100 100

Diethylstilbestrol (DES) 236 221 117 69
Tamoxifen 4 3 6 2
Coumestrol 20 140 102 98

Isoflavones Genistein 4 87 198 182

Daidzein 0.1 0.5 97 80
Formononetin <0.01 <0.01 6 2
Biochanin A <0.01 <0.01 36 53
Ipriflavone <0.01 <0.01 11 3

Endocrine disruptors Bisphenol A 0.01 0.01 50 41

o,p -DDT 0.01 0.02 54 10
Nonylphenol 0.05 0.09 62 34
Methoxychlor <0.01 <0.01 9 2

The RBA of each competitor was calculated as the E2:competitor concentration ratio required to reduce the specific radioligand binding by 50% (ratio of IC50 values). The RBA
value for E2 was arbitrarily set at 100. The RTA of each compound was calculated as the ratio of the luciferase reporter gene induction value, at a concentration of 1 ␮M of
the relevant compound, relative to the luciferase reporter gene induction value using 17␤ estradiol at 1 ␮M. The transactivation activity of 17␤ estradiol was arbitrarily set at
100. Data adapted from Kuiper et al. (1998).

with preferential binding of genistein to ER␤ (Table 1). Specific
binding affinity to ERs enables isoflavones to elicit both estrogenic
and antiestrogenic effects depending on the tissue, as well as on
isoflavone and endogenous estradiol levels. For instance, in var-
ious cell lines, in the presence of physiological levels (1 nM) of
estradiol (E2), genistein acts as an anti-estrogen whereas at lev-
els of E2 found in postmenopausal women (0.01 nM), genistein
shows additive agonistic effects (Hwang et al., 2006). Genistein
also has ER-mediated biphasic effects in intestinal cell prolifera-
tion (Chen and Donovan, 2004). Thus, isoflavones, as stress-induced
phytochemicals, may provide health benefits through their selec-
tive estrogen receptor modulator (SERM) activities.
6. Role of estrogens in metabolism
Studies in humans and rodents have shown that ERs are impor-
tant mediators of the action of estrogen on lipid and glucose
metabolism. Estrogens have been reported to affect adiposity either
directly, by modulating lipogenesis, lipolysis or adipogenesis, or
indirectly, by modulating appetite or energy expenditure (Cooke
and Naaz, 2004). As mentioned above, the concept of estrogens
modulating metabolic features derived originally from the obser-
vation that postmenopausal women develop visceral obesity and
insulin resistance as a result of low levels of estrogens. Interest-
ingly, hormonal replacement therapy normalizes these symptoms
(Ahmed-Sorour and Bailey, 1980; Bailey and Ahmed-Sorour, 1980).
Estrogens also play an important role in glucose homeostasis, and
are known to modulate insulin sensitivity (Godsland, 2005). Vari-
ations in human glucose homeostasis are observed during the
menstrual cycle, and diabetes becomes more resistant to treatment
during the luteal phase (Case and Reid, 2001). Relevant genetic
evidence that estrogen modifies metabolism can also be found
in humans: individuals with mutations in the aromatase gene, an
enzyme that converts androgens into estrogens, display truncal
obesity, insulin resistance and hyperlipidemia (Carani et al., 1997).
Studies in rodents are far more advanced, and confirm the impor-
tance of ER␣ in modulating both lipid and glucose metabolism. Mice
lacking either ER␣ or the aromatase gene have increased adipos-
ity and fatty livers (Heine et al., 2000; Jones et al., 2000; Takeda
et al., 2003), confirming the role of estrogens in the regulation
of adiposity. In aromatase knockout mice (ArKO), liver steatosis
is associated with a decreased expression of genes responsible
for fatty acid oxidation, but can be normalized with E2 treatment
(Nemoto et al., 2000). The function of ER␤ is less clear—to date no
overt metabolic phenotype has been described in mice lacking this
receptor (␤ERKO), under normal dietary conditions (Ohlsson et al.,
2000). Recently, it was found that, when challenged with a high fat
diet, ␤ERKO mice increase their weight and adiposiy to a greater
extend than do wild-type mice (Foryst-Ludwig et al., 2008).
Estrogens have also been shown to modulate glucose
metabolism in rodents. For instance in models of type 2 diabetes,
ovariectomy induces hyperglycemia, whereas in males, estrogen
perfusion reverses diabetes (Louet et al., 2004). Similarly, mice lack-
ing either functional ER␣ or aromatase display glucose intolerance
and insulin resistance (Heine et al., 2000; Jones et al., 2000; Takeda
et al., 2003). Indeed, ER␣ and ER␤ are both expressed in muscle
cells and are known to modulate the expression of the glucose
transporter 4 (Glut4), a key molecule for the import of glucose in
cells (Barros et al., 2006). The endocrine pancreatic function is also
directly modulated by estrogen levels. In rats, circulating insulin
levels have been shown to vary during the estrous cycle (Bailey
and Matty, 1972). Estrogens and endocrine disruptors such as
Bisphenol-A (BPA) have been reported to modulate insulin content
in ␤-cells via ER␣, but not via ER␤ (Alonso-Magdalena et al., 2008).
These molecules also protect ␤-cells from oxidative stress, apopto-
sis, and streptozotocin-induced injuries in both genders, through
ER␣ (Le May et al., 2006). In addition, several studies reported that
estrogens increase glucose-stimulated insulin secretion in isolated
islets (Nadal et al., 1998; Adachi et al., 2005; Alonso-Magdalena et
al., 2006, 2008; Le May et al., 2006; Martensson et al., 2008).
Finally, estrogens have been reported to modulate energy home-
ostasis indirectly, through the central nervous system (CNS) and
the hypothalamus. By sensing endocrine and metabolic signals, the
hypothalamus engages distinct effector pathways, which result in
behavioral, endocrine and metabolic changes, to maintain energy
homeostasis. Several hormones are capable of influencing energy
intake and expenditure. For instance, leptin and insulin modulate
the activity of hypothalamic neurons in the arcuate nucleus, ulti-
mately leading to changes in food intake (Schwartz et al., 2000). It
has been shown recently that estrogens are among the hormones
that modulate the energy balance. Deficiency in aromatase or ER␣,
or ovariectomy, decreases physical activity and energy expendi-
ture (Heine et al., 2000; Cooke et al., 2001; Meli et al., 2004),
whereas treatment with E2 increases locomotor activity through
an ER␣-dependent mechanism (Ogawa et al., 2003). Both estro-
gen receptors are found in the hypothalamic nuclei and modulate
34 C.R. Cederroth, S. Nef / Molecular and Cellular Endocrinology 304 (2009) 30–42
food intake (Liang et al., 2002) and locomotor activity (Ogawa et
al., 2003). Specific silencing of ER␣ in the hypothalamus of female
rodents leads to similar phenotypes to those observed in consti-
tutive mutant mice, including obesity, hyperphagia and reduced
energy expenditure (Musatov et al., 2007). Although it remains
unknown whether or not these phenotypes are associated with
changes in the expression of hypothalamic neuropeptides, these
results show that hypothalamic ER␣ is essential in the regulation of
energy balance. Supporting a direct effect of E2 on the hypothalamic
control of adiposity, it was shown that E2 modulates hypothalamic
synapticity by bypassing the leptin receptor to act directly on the
downstream Stat3 signaling in the hypothalamus, thus decreasing
body weight (Gao et al., 2007).
Overall, estrogens appear to be crucial regulators of metabolic
functions by directly and indirectly (via the CNS) modulating energy
homeostasis. Whether or not dietary soy and phytoestrogens have
effects on energy homeostasis through estrogen-mimics is still a
matter of controversial debate, but is clearly a plausible hypothesis.
7. Effects of soy protein and phytoestrogens on human
metabolism
The low frequency of obesity and related metabolic disorders
in Asian populations has drawn attention towards soy, which is a
characteristic component in asiatic diets. We searched the PubMed
literature database for epidemiological and clinical studies evaluat-
ing the effects of soy or isolated isoflavones on human metabolism,
and the main results are summarized in Tables 2–5.
Epidemiological studies have shown that type-2 diabetes is four
times less prevalent in Japanese people in Tokyo than in Japanese-
Americans in Seattle (Fujimoto et al., 1987, 1991). Consumption of
more than 12.6 grams of soy protein per day is associated with a
lower risk of glycosuria, a strong predictor of diabetes (Yang et al.,
2004). Similarly, several studies have reported that isoflavone con-
sumption by postmenopausal women correlated with lower body
mass index (BMI), and higher HDL levels (Guthrie et al., 2000;
Goodman-Gruen and Kritz-Silverstein, 2001, 2003).
Clinical studies also suggest that soy protein or isoflavones may
improve metabolic parameters. For instance, a metaanalysis of 38
trials (Anderson et al., 1995) as well as more recent reports (Crouse
et al., 1999; Takatsuka et al., 2000; Teixeira et al., 2000; Gardner et
al., 2001; Jayagopal et al., 2002; Greany et al., 2004) demonstrated
a significant reduction in plasma concentrations of total and LDL
cholesterol in humans exposed to soy proteins. In addition, post-
menopausal Japanese women treated for 24 weeks with isoflavones
exhibited a lower fat mass (Wu et al., 2006). Obese patients treated
with soy protein isolates for 12 weeks had lower body weight and
BMI, with decreased cholesterol and LDL levels in the blood (Allison
et al., 2003). Additionally, a 6-month clinical trial was conducted
Table 2
Epidemiological studies evaluating the effects of soy or isoflavones on human metabolism
Model Number of Dose
individuals (total)
Pre and postmenopausal women 323 Soy protein intake >12.61 g/day
Pre and postmenopausal women 944 Fermented soy bean
Postmenopausal women 208 Genistein intake >1 mg/day
Postmenopausal women 939 Isoflavone intake >0.236 mg/day
In these studies, serum isoflavone levels were not evaluated. Abbreviations: W, weight; F/BMI, fat or body mass index; G, serum glucose; I, serum insulin; TC, total cholesterol;
LDL, low density lipoprotein; HDL, high density lipoprotein; TG, triglycerides; FFA, free fatty acids; ND, not determined. Parameters listed here that do not appear in the table
were not analyzed in the article in question.
to compare the effects of isoflavones with that of conjugated estro-
gens on blood glucose, insulin, and lipid profiles in postmenopausal
Taiwanese women. The study revealed that during fasting both glu-
cose and insulin levels were significantly reduced by soy isoflavones
(100 mg/day) and conjugated estrogens (0.625 mg/day) (Cheng et
al., 2004).
In contrast to the above mentioned trials, a significant number
of studies reported an absence of beneficial effects of soy on classi-
cal metabolic parameters such as body weight, serum lipid profiles,
fat mass, blood glucose and insulin profiles (Yamashita et al., 1998;
Anderson and Hoie, 2005; Li et al., 2005; Hall et al., 2006; Ikeda
et al., 2006; Anderson et al., 2007). These discrepancies make it
difficult to draw firm conclusions regarding the beneficial effect
of soy on glucose and lipid metabolism. When comparing these
different clinical trials, the underlying causes of conflicting results
are probably related to the variability of experimental designs and
exposition protocols (route of administration, composition, dose,
and duration), the capacity of individuals to produce equol and
the genetic susceptibility. Clearly more standardized studies are
needed to further evaluate these putative beneficial effects.
8. Actions of soy on metabolism in rodents
The current scientific evidence concerning the role of soy and
isoflavones in rodents is based on studies where animals have been
exposed either to purified isoflavones (injected or supplemented
in the diet itself) or soy protein isolates (SPIs) (for a complete
list of studies, see Tables 6–10). The difficulty in analyzing SPI
studies arises from the fact that information on isoflavone levels
and composition are quite often incomplete, making interpreta-
tions problematic and comparisons hazardous. On the other hand,
supplementation of the diet with soy proteins is more relevant
than injection-based studies. This does, however, raise questions
as to which compound is responsible for the observed effects and
whether the relative benefits of soy are not in fact due to the poor
performance of casein itself, which is usually used as control pro-
tein.
In comparison to human studies which mainly focus on serum
lipid analysis because of the clinical importance of atherosclero-
sis and the risk in cardiovascular diseases, reports in rodents are
rather oriented towards the assessment of soy-derived compounds
on weight and fat loss, and in fewer cases, insulin sensitivity. Still
concerning the effects of soy or isoflavones on serum lipid profiles,
most rodent studies that have assessed these parameters under
healthy or diabetic states point towards an improvement in total
cholesterol (TC) and/or triglyceride (TG) levels after consumption of
dietary soy, SPI, isoflavones or genistein (Kirk et al., 1998; Nagasawa
et al., 2003; Ae Park et al., 2006; Penza et al., 2006; Cederroth et al.,
2008; Nordentoft et al., 2008; Torre-Villalvazo et al., 2008). Only
Metabolic Effects References
For >12.61 g/day: Lower risk in glycosuria in Yang et al. (2004)
postmenopausal women with BMI
<25 kg/m2
No effects on W or F/BMI Ikeda et al. (2006)
No effects on G, I, TC, LDL, TG Goodman-Gruen and
Kritz-Silverstein (2001)
Lower F/BMI, increased HDL
No effects on F/BMI, TC, LDL, HDL, Lower TG de Kleijn et al. (2002)
 C.R. Cederroth, S. Nef / Molecular and Cellular Endocrinology 304 (2009) 30–42 35

Table 3

Clinical studies performed using dietary supplementation with soy protein isolates (SPI)

Model Number of Duration SPI and isoflavones intake Serum isoflavone Metabolic effects References
individuals

Young healthy normolipidemic Total = 22 13 days High isoflavone SPI Genistein: 0.75 No effects on TC, LDL, Sanders et al. (2002)
men and women (56 mg/day) vs low TG
isoflavone SPI (2 mg/day)
Daidzein: 0.3 Increased HDL
Equol: 0.1

Obese women 22 (21) 16 weeks SPI (15 g/day) ND No effects on W, Anderson et al. (2007)
(150 mg/day isoflavones) F/BMI, G, I, TC, LDL,
vs casein (15 g/day) HDL, TG
(10 mg/day isoflavones)
Postmenopausal women 100 (102) 12 months SPI (25.6 g/day) ND No effects on F/BMI Kok et al. (2005)
(99 mg/day isoflavones)
vs milk
Moderate 31 (33) 12 weeks SPI with isoflavone ND No effects on W, Gardner et al. (2001)
hypercholesterolemic (80 mg) or without vs F/BMI, HDL, TG
postmenopausal women milk (42 g/day)
Lower TC, LDL

Moderate Total = 146 9 weeks SPI (25 g/day) (62 mg/day ND No effects on HDL, TG Crouse et al. (1999)
hypercholesterolinemic men isoflavones) vs Casein
and women (25 g/day)
Lower TC, LDL

Normocholesterolinemic 71 (72) 6 weeks SPI with isoflavones ND No effects on W, Greany et al. (2004)
(44.3 mg/day) vs milk F/BMI, TC, LDL, HDL,
TG
Mildly hypercholesterolinemic 71 (72) 6 weeks SPI with isoflavones ND No effects on W, F/BMI Greany et al. (2004)
(44.3 mg/day) vs milk
Lower TC, LDL, TG
Increased HDL

Obese men and women 50 (50) 12 weeks SPI vs control ND No effects on HDL Allison et al. (2003)
Lower W, BMI, TC, LDL

Overweight and obese men Total = 90 12 weeks SPI vs milk ND No effects on W, G, TC, Anderson and Hoie (2005)
and women LDL, HDL, TG
Obese male and women 17 (19) 16 weeks SPI vs meat ND No effects on W, Yamashita et al. (1998)
F/BMI, G, TC, LDL, HDL,
TG
Obese men and women 46 (36) 3 or 6 months SPI vs individual dietary ND No effects on I, TC, Li et al. (2005)
intervention LDL, HDL, TG
Lower W, F/BMI, G
12 months SPI vs individual dietary ND No effects on W, Li et al. (2005)
intervention F/BMI, G, I, TC, LDL,
HDL, TG

Abbreviations: SPI, soy protein isolate; W, weight; F/BMI, fat or body mass index; G, serum glucose; I, serum insulin; TC, total cholesterol; LDL, low density lipoprotein; HDL,
high density lipoprotein; TG, triglycerides; FFA, free fatty acids; ND, not determined. Parameters that do not appear in the table were not analyzed in the article in question.
Serum isoflavone values are expressed in ␮M. For control groups, the number of individuals (n) is shown in parentheses.

one study shows an absence of effects on total cholesterol and TG
after consumption of SPI (Aoyama et al., 2000a,b).
As mentioned above, most studies have evaluated the effects
of soy-derived compounds on adiposity. For instance, Long-Evans
rats or ovariectomized (OVX) ddY mice fed with a soy-rich diet
have reduced weight and less adipose deposition than those fed
on a soy-free diet (Wu et al., 2004; Bu and Lephart, 2005). Simi-
larly, Long-Evans rats or CD-1 male mice fed with dietary soy (with
serum levels of genistein and daidzein reaching 0.5 ␮M and equol
about 10 ␮M) have 50% decreased adipose weight (Lephart et al.,
2004a; Cederroth et al., 2007). In addition, dietary supplementa-
tion with 500–1500 ppm of genistein, with a serum equivalent of
about 2 ␮M, decreases fat pad weights by 50% in C57/BL6 mice (Naaz
et al., 2003). The effects on adiposity are dose-dependent, since the
magnitude of adipose weight reduction correlates with increasing
doses of soy-derived phytoestrogens (Lephart et al., 2004b). Simi-
larly, subcutaneous injections of genistein (8–200 mg/kg/day) for 21
days in C57/BL6 ovariectomized mice decrease adipose gain (Naaz
et al., 2003). In contrast, in a study where male mice were exposed
to daily oral doses of genistein of up to 50 mg/kg/day, fat mass was
increased. However fat mass decreased at doses of 200 mg/kg/day
(Penza et al., 2006). Overall, these results suggest that high phar-
macological doses of phytoestrogens yielding serum levels in the
micromolar range inhibit adipose tissue deposition.
The mechanisms by which dietary soy and phytoestrogens
reduce adiposity are unclear. Recently it was found that increased
energy expenditure and locomotor activity coupled with a marked
shift towards the use of lipids as fuel source were two important
contributing factors for the leanness observed in mice exposed to
dietary phytoestrogens (Cederroth et al., 2007). This decrease in
fat abundance correlates with an increased activation, in periph-
eral tissues, of the AMP-activated protein kinase (AMPK) and its
downstream target Acetyl-CoA carboxylase (ACC) (Cederroth et al.,
2008), two enzymes which are key regulators of fatty acid oxida-
tion (Winder and Hardie, 1996; Ruderman and Prentki, 2004). In
these mice, two targets of AMPK, peroxisome proliferator activated
receptor (PPAR)␥ co-activator (PGC)-1␣ and (PPAR)␣, which regu-
late mitochondrial and peroxisomal metabolism respectively (Zong
et al., 2002; Lin et al., 2005), were preferentially upregulated in
adipose tissue (Cederroth et al., 2008). Additionally, in vitro expo-
sure of cultured 3T3-L1 adipocytes to genistein has been reported
to significantly activate AMPK and ACC (Hwang et al., 2005). Fur-
36 C.R. Cederroth, S. Nef / Molecular and Cellular Endocrinology 304 (2009) 30–42

Table 4

Clinical studies performed using dietary supplementation with phytoestrogens or isoflavones

Model Number of Duration Isoflavone intake Serum or urinary isoflavone Metabolic effects References
individuals

Postmenopausal women Total = 136 6 months Isoflavones Serum genistein: 0.32 (0.28) No effects on TC, TG Wu et al. (2006)
(75 mg/day) vs
placebo
Daidzein: 0.89 (0.27) Lower F/BMI
Equol: 0.19 (0.07) Increased HDL
Glycitein: 0.19 (0.06)

Postmenopausal women Total = 34 2 months Isoflavones Urinary genistein: 12.2 (0.65)

(50 mg/day) vs
placebo
Daidzein: 8.1 (0.22)
Equol: No effects on W, Weickert et al. (2006)
F/BMI, G, I
- Equol producers: 5.8 (0.237)
- Non-producers: 0.245 (0.160)

Healthy Postmenopausal women Total = 117 8 weeks Isoflavones vs Urinary genistein: 7.27 (0.42) No effects on G, I, TC, Hall et al. (2006)
placebo LDL, HDL, TG
Daidzein: 5.76 (0.22)
Equol:
- Equol producers: 2.61 (0.13)
- Non-producers: 0.15 (0.094)

Postmenopausal women 38 (40) 3 months Isoflavones ND No effects in TC, HDL Dalais et al. (2003)
(118 mg/day) vs
placebo
Lower LDL, TG

Postmenopausal women with Total = 32 12 weeks Phytoestrogens ND No effects on W, G, Jayagopal et al. (2002)
type 2 diabetes vs placebo HDL, TG
Lower I, TC, LDL

Abbreviations: SPI, soy protein isolate; W, weight; F/BMI, fat or body mass index; G, serum glucose; I, serum insulin; TC, total cholesterol; LDL, low density lipoprotein; HDL,
high density lipoprotein; TG, triglycerides; FFA, free fatty acids; ND, not determined. Parameters that do not appear in the table were not analyzed in the article in question.
Serum isoflavone values are expressed in ␮M. Urinary isoflavone values are expressed in mM. For control groups the number of individuals (n) and the corresponding serum
or urinary isoflavone values are shown in parentheses.

thermore, in vitro studies using isolated rat adipocytes have shown
that genistein stimulates lipolysis through the inhibition of cAMP
phosphodiesterases (Szkudelska et al., 2000). Similarly, it has been
recently shown that an isoflavone-free peptide mixture from black
soybean (BSP) significantly activates both AMPK and ACC in C2C12
myocytes, in a dose dependent manner (Jang et al., 2008). Interest-
ingly, a high fat diet containing 10% of this same BSP fed to mice
for 13 weeks, led to AMPK activation in WAT of mice compared to
control animals.
Fewer studies have evaluated the effects of soy and phytoe-
strogens on glucose homeostasis. Rats fed with SPI exhibit lower
glucose and insulin levels in the plasma (Hurley et al., 1998) as well
as increased peripheral insulin sensitivity (Lavigne et al., 2000).
In rat models of obesity and type 2 diabetes (SHR/N-cp rats and
ZDFxSHHF rats), treatment with isoflavones decreases glycemia or
circulating insulin levels (Ali et al., 2005; Davis et al., 2005). Sim-
ilarly, in mice models of obesity and diabetes, treatment with soy
protein or genistein lowers glycemia (Aoyama et al., 2000a; Ae Park
et al., 2006).
Potential mechanisms by which dietary soy may improve
glucose metabolism have been recently proposed. Dietary soy
improves insulin sensitivity by increasing glucose uptake prefer-
entially in skeletal muscles (Cederroth et al., 2008). This improved
insulin responsiveness in mice exposed to dietary soy may take
place, at least in part, by an improvement of the PI(3)K-Akt sig-
naling by the dietary soy-activated AMPK (Cederroth et al., 2008).
Whether these effects are due to soy proteins or phytoestrogens
remains unknown.
The effects of soy and phytoestrogens on the pancreas are less
known. Recently, an in vivo study has evaluated ␤-cell function
after exposure of CD-1 male mice to dietary soy. Dietary soy lowers
pancreatic insulin content, while high glucose-stimulated insulin
secretion is unchanged, suggesting that insulin secretion is ame-
liorated (Cederroth et al., 2008). Although it is not clear whether
these beneficial effects are due to the isoflavones themselves or
other components of soy proteins, recent reports indicate that the
isoflavone genistein may itself have a direct beneficial effect on
pancreatic ␤-cells. For example, genistein has been shown to stim-

Table 5

Clinical studies performed using dietary supplementation with ␤-conglycitin

Model Number of individuals Duration Design Metabolic effects References

Hyperlipidemic male and women 69 (69) 12 weeks Beta-conglycinin vs No effects on W, F/BMI, G, I, TC, Kohno et al. (2006)
casein LDL decreased TG, FFA
increased HDL
Obese male and women 24 (22) 12 weeks Beta-conglycitnn vs No effects on TC, LDL, HDL, TG Kohno et al. (2006)
casein lower W, F/BMI, FFA

Abbreviations: SPI, soy protein isolate; W, weight; F/BMI, fat or body mass index; G, serum glucose; I, serum insulin; TC, total cholesterol; LDL, low density lipoprotein; HDL,
high density lipoprotein; TG, triglycerides; FFA, free fatty acids; ND, not determined. Parameters that do not appear in the table were not analyzed in the article in question.
For control groups, the number of individuals (n) is shown in parentheses.
Table 6

Animal studies performed using dietary supplementation with dietary soy

Species Duration (days) Dose (ppm) Total serum Number of Weight Food intake Fat mass Glycemia Circulating insulin Reference
isoflavone levels animals

Male rats
Long-Evans Life long 600* ∼7.07 ND Decreased Increased Decreased ND Decreased Lephart et al. (2004a)
Sprague–Dawley Life long 600* ∼8.7 57 (57) Decreased No effect ND ND ND Weber et al. (2001)

Female OVX rats

OVX Long-Evans 42 days 600* ∼3.3 ND Decreased Increased Decreased ND ND Bu et al. (2005)

Male mice
CD-1 Life long 600* ∼10 12 (12) Decreased Increased Decreased ND ND Cederroth et al. (2007)
CD-1 Life long 600* ∼10 5–14 Decreased ND Decreased No effect Decreased a , b Cederroth et al. (2008)

C.R. Cederroth, S. Nef / Molecular and Cellular Endocrinology 304 (2009) 30–42
(5–19)
Female mice
CD-1 Life long 600* ∼10 12 (12) Decreased ND Decreased ND ND Cederroth et al. (2007)

Studies using dietary soy are distinguished from SPI in that they include analysis of serum isoflavone abundance, while all the presented studies in rodents using SPI, presence or absence of isoflavones has not been measured. For
clarity, data on serum lipid profiles are not presented. Abbreviations: OVX, ovariectomized; ND, not determined. Symbols: * Total isoflavone content.
a
Increased glucose tolerance.
b
Increased insulin sensitivity.
Total serum isoflavone values are expressed in ␮M. For control groups, the number of individuals (n) is shown in parentheses.

Table 7

Animal studies performed using dietary supplementation with soy protein isolates (SPI)

Species Duration (days) Dose (% SPI) Number of animals Weight Food intake Fat mass Glycemia Circulating insulin Reference
(controls)

Male rats
Sprague–Dawley 28 22.6 10 (10) Decreased ND Decreased Decreased Decreased Hurley et al. (1998)
Wistar 28 22.9 20 (20) No effect No effect ND Decreased Decreased a , b Lavigne et al. (2000)
Wistar 10 22.4 8 (8) No effect No effect No effect No effect No effect Nagasawa et al. (2003)
Sprague–Dawley 28 41.1 8 (6) No effect No effect No effect Decreased ND Aoyama et al. (2000b)
Sprague–Dawley 180 30 8 (8) Decreased ND No effect No effect No effect Torre-Villalvazo et al. (2008)
Sprague–Dawley on a HFD 180 30 8 (8) Decreased ND Decreased No effect No effect Torre-Villalvazo et al. (2008)

Male mice
Obese yellow KK-Ay 14 41.6 10 (10) No effect No effect Decreased No effect ND Aoyama et al. (2000a)
Obese yellow KK-Ay 28 41.1 12 (12) No effect ND Decreased ND ND Aoyama et al. (2000b)
Obese yellow KK-Ay 63 50 10 (10) Decreased No effect Decreased Decreased Decreased Nordentoft et al. (2008)
CD-1 Life long ND# 10 (13) Decreased Increased Decreased ND ND a Ruhlen et al. (2008)

Female mice
CD-1 Life long ND# 10 (13) Decreased ND Decreased ND ND Ruhlen et al. (2008)

For clarity, data on serum lipid profiles are not presented. In the following studies, serum isoflavone levels were not analyzed. Abbreviations: OVX, ovariectomized; SPI, soy protein isolate; ND, not determined.
a
Increased glucose tolerance.
b
Increased insulin sensitivity.
#
Estrogenic activity tested in vitro.
For control groups, the number of individuals (n) is shown in parentheses.

37
38 C.R. Cederroth, S. Nef / Molecular and Cellular Endocrinology 304 (2009) 30–42

Dolinoy et al. (2006)

Dolinoy et al. (2006)

ulate insulin secretion in the insulin-secreting cell lines INS-1 and

Ae Park et al. (2006)

Penza et al. (2006)

Naaz et al. (2003)

Naaz et al. (2003)
Naaz et al. (2003)
Naaz et al. (2003)
MIN6 and in mouse pancreatic islets, at nano- and micromolar con-
centrations (Liu et al., 2006). In addition, genistein has been also
reported to prevent cytokine-induced cell damage in rat insuli-
Reference

noma cells (RIN cells) (Kim et al., 2007). Genistein and daidzein have
been shown to prevent diabetes onset in non-obese diabetic (NOD)
mice by preserving pancreatic ␤ cell function (Choi et al., 2008).
Similarly, Lee (2006) recently reported that in streptozotocin-
induced diabetic rats, supplementation with genistein (600 mg/kg
Circulating

No effectc

of diet) cause a 2 fold increase in plasma insulin levels. Collectively,

insulin

NDa,b

these data suggest that dietary soy and phytoestrogens may have
ND

ND

ND
ND
ND
ND
therapeutic significance in reducing the severity of diabetes, and
improving ␤-cell survival and function. However, further research
is needed, first to confirm and then to analyze in detail, how phy-
Decreased
Glycemia

toestrogens exert anti-hyperglycemic actions through pancreatic

␤-cells.
ND

ND

ND

ND
ND
ND
ND

9. Central actions of phytoestrogens

Decreased
Decreased
Decreased
Increased

Whether or not soy-derived compounds modulate energy

No effect
Fat mass

expenditure via the CNS is unclear, but recent data suggest that
ND
ND

ND

some effects may be centrally mediated. In mice or rats fed soy-rich

For clarity, data on serum lipid profiles are not presented. Abbreviations: OVX, ovariectomized; ND, not determined. Symbols: * Total isoflavone content.

diets, feeding behavior and locomotor activity were significantly

affected, suggesting that the central regulation of energy balance
Food intake

in the hypothalamus may be modulated by soy or phytoestro-

No effect
No effect

gens (Lephart et al., 2003; Cederroth et al., 2007). For example, in

mice exposed to dietary soy, the increased locomotor activity and
ND

ND

ND
ND
ND
ND

preferential use of lipids as fuel source is associated with a 40%

decrease in mRNA levels of AgRP in the hypothalamus (Cederroth
et al., 2007). AgRP is a key neuropeptide regulating energy expen-
Decreased

Decreased
No effect
No effect

diture in the arcuate nucleus of the hypothalamus (Stutz et al.,

Serum genistein values are expressed in uM. For control groups, the number of individuals (n) for is shown in parentheses.
Weight

2005). Mice lacking AgRP, or having 50% reduction of AgRP lev-

ND
ND
ND
ND

els in the hypothalamus, display an increased metabolic rate, lipid

utilization and locomotor activity (Makimura et al., 2002; Wortley
et al., 2005). In a similar study, rats exposed to the same soy-based
Number of

diet exhibited higher expression of transcripts for NPY (an approx-

animals

10 (10)

imately 40% increase) in the arcuate and paraventricular nuclei of

4 (4)
4 (4)
4 (4)
4 (4)

the hypothalamus. NPY is an orexigenic neuropeptide known to

ND

ND
20

stimulate food intake both in rodents and humans (Schwartz et al.,

2000). Recently, it has been reported that an isoflavone-free pep-
tide mixture from black soybean (BSP) significantly decreased food
Serum genistein

intake in rats and leptin-deficient ob/ob mice (Jang et al., 2008).

The authors presented evidence that BSP has anorectic effects in
2.55 (0.08)
3.81 (0.08)
1.79 (0.08)
1.02 (0.08)

association with the induction of hypothalamic STAT3 phosphory-

levels

lation, a major pathway involved in suppression of food intake and

0.09
ND
ND

ND
Animal studies performed using dietary supplementation with genistein

energy expenditure (Bates and Myers, 2003). Further studies are

needed to ascertain whether or not soy peptides or dietary phy-
toestrogens act directly on the hypothalamus to improve lipid and
Dose ppm

glucose metabolism.
800*
2000*
250*

250*

300*
500*
1000*
1500*

10. Potentially adverse effects in consuming soy and

soy-derived phytoestrogens
18, during gestation

18, during gestation

The extent to which soy food and its bioactive component

Duration (days)

genistein pose potential health risks is still a matter of debate

(Setchell, 2006). In fact, the initial recognition and identification
No effects on insulin sensitivity.
No effects on glucose tolerance.

of phytoestrogens as bioactive compounds was made in the 1940s

12 days

Increased glucose tolerance.

when it was found that formononetin, an isoflavone present in red

42
15

clover (Trifolium pratense L.), caused a devastating infertility syn-

drome in sheep grazing in clover pasture (Bennetts et al., 1946).
In another study, high levels of phytoestrogens were found in the
Female OVX Mice
Db/Db-C57BL/KSJ

leaves of stunted desert annuals in a dry year, leading ultimately

Female Mice

to impaired reproduction when ingested by the California quail

Male mice

(Lophortyx californicus). In wet years these quails bred normally and

C57BL/6

C57BL/6
Species
Table 8

phytoestrogens were largely absent in these herbs (Leopold et al.,

Avy /a

Avy /a

1976).
 C.R. Cederroth, S. Nef / Molecular and Cellular Endocrinology 304 (2009) 30–42 39

Table 9

Animal studies performed using subcutaneous injections or gavage of genistein

Species Duration Dose Serum Number of Weight Food intake Fat mass Reference
(days) genistein levels animals

Subcutaneous injections
Female OVX mice
C57BL/6 21 20 mg/kg/day ND 10 (10) No effect No effect No effect Naaz et al. (2003)
80 mg/kg/day ND 10 (10) No effect No effect Decreased Naaz et al. (2003)
200 mg/kg/day ND 10 (10) No effect No effect Decreased Naaz et al. (2003)

Oral gavage
Male mice
C57BL/6 mice 15 50 ␮g/kg 35 (28) 20 (20) No effect No effect No effecta,b Penza et al. (2006)
500 ␮g/kg 66 (28) 20 (20) No effect No effect Increaseda,b Penza et al. (2006)
5,000 ␮g/kg 74 (28) 20 (20) No effect No effect Increaseda,b Penza et al. (2006)
50,000 ␮g/kg 98 (28) 20 (20) No effect No effect Increaseda,b Penza et al. (2006)
200,000 ␮g/kg 223 (28) 20 (20) Decreased Decreased Decreased a,c Penza et al. (2006)

Female mice
C57BL/6 15 50 ␮g/kg 35 (28) 20 (20) No effect No effect No effecta,b Penza et al. (2006)
500 ␮g/kg 66 (28) 20 (20) No effect No effect No effecta,b Penza et al. (2006)
5,000 ␮g/kg 74 (28) 20 (20) No effect No effect No effecta,b Penza et al. (2006)
50,000 ␮g/kg 98 (28) 20 (20) No effect No effect No effect a,b Penza et al. (2006)
200,000 ␮g/kg 223 (28) 20 (20) Decreased Decreased Decreaseda,c Penza et al. (2006)

For clarity, data on serum lipid profiles are not presented. In the following studies, serum glucose and insulin were not analyzed. Abbreviations: OVX, ovariectomized; ND, not
determined.
a
No effects on glucose tolerance.
b
No effects on insulin sensitivity.
c
Increased insulin sensitivity.
Serum genistein values are expressed in nM. For control groups, the number of individuals (n) and the corresponding serum genistein values are shown in parentheses.

Numerous studies in rodents clearly show that purified genis-
tein (delivered by subcutaneous or intraperitoneal injections) has
detrimental effects. The incidence of uterine carcinoma increased
by 31% in neonatal mice treated subcutaneously with 50 mg/kg/day
of genistein for a 5-day period (Newbold et al., 2001). Female mice
that received subcutaneous injections of 50 mg/kg/day postnatally,
while remaining fertile, could not deliver pups and had multi-
oocyte follicles (MOFs) (Jefferson et al., 2005, 2007). In adult male
rats, exposure to dietary soy decreased androgen levels and prostate
weight (Weber et al., 2001). Finally, it has been shown in female
mice that dietary phytoestrogens accelerated the time of vaginal
opening in immature CD-1 mice (Thigpen et al., 2003). However,
with the exception of dietary soy experiments, most of these find-
ings may have little relevance to humans consuming soy food, in
particular because the route of administration differs (oral versus
injection) resulting in the bypass of the intestines, which provide
a limiting barrier to the bioavailability of isoflavones. This is par-
ticularly relevant since the two main isoflavones, genistein and
daidzein, are present in soy as ␤-D-glycosides, namely genistin and
diadzin (Fig. 1). Both genistin and daidzin are biologically inactive
and require the hydrolyzation of the glycosidic bond by glucosi-
dases of the intestinal bacteria to produce the biologically active
aglycone forms.
In humans, the use of soy or purified phytoestrogens in women
at high risk of, or diagnosed with, breast cancer as well as in infants
fed with soy-based formula are legitimate areas of concern. Con-
cerning breast cancer, caution is warranted since the available data
are conflicting, and there is evidence for both inhibitory and stim-
ulatory effects of dietary soy on breast cancer cell growth (Duffy
et al., 2007). In vivo data from animals suggest that genistein may
interfere with the inhibitory effect of tamoxifen on breast cancer
cell growth (Ju et al., 2002; Liu et al., 2005), while epidemiolog-
ical studies in humans show that exposure in early childhood or
early adolescence protects against the development of breast can-
cer as an adult (Wu et al., 2002). Concerning soy-based formula,
caution should prevail, even though it has been consumed by mil-
lions of infants over the past decades without apparent detrimental
effects. Infancy is a very sensitive period for endocrine disruption,
and exposure to significant levels of phytoestrogens may ultimately
lead to adult onset diseases.
11. Summary and conclusion
Current evidence from animal and human studies suggests that
diets rich in soy and phytoestrogens have beneficial effects on many
aspects of diabetes and obesity. In animal studies, soy and phy-
toestrogens are effective at reducing adipose tissue and improving
glucose uptake. However, available data from human studies do not
offer clear support, and further research is required before a firm
conclusion can be made about the benefits of soy and phytoestro-

Table 10

Animal studies performed using dietary supplementation with soy proteins (isoflavone-free)

Species Duration Dose (%) Number of Weight Fat mass Glycemia Circulating Reference
(days) animals insulin

ICR 28 23.7 (␤-conglycinin) 10 (10) Decreased No effect Decreased Decreased Moriyama et al. (2004)
ICR 28 21.9 (glycinin) 10 (10) Decreased No effect No effect No effect Moriyama et al. (2004)
yellow KK-Ay 28 23.7 (␤-conglycinin) 10 (10) Decreased No effect No effect Decreased Moriyama et al. (2004)
yellow KK-Ay 28 21.9 (glycinin) 10 (10) Decreased No effect No effect No effect Moriyama et al. (2004)
Obese C57BL/6 91 Black soy protein 9 (9) Decreased No effect ND ND Jang et al. (2008)

For clarity sakes, data on serum lipid profiles are not presented. Food intake was not measured in these studies. In the following studies, serum glucose and insulin were not
analyzed. For control groups, the number of individuals (n) is shown in parentheses.
40 C.R. Cederroth, S. Nef / Molecular and Cellular Endocrinology 304 (2009) 30–42
gens in the context of adiposity control and glucose metabolism.
The specific soy protein components that may lead to metabolic
improvements have yet to be determined. Phytoestrogens appear
to have beneficial actions both on glucose and lipid metabolism
but additional micronutriments such as saponins, phytosterols,
trypsin inhibitors, as well as amino acid and protein composi-
tion, may have additive or synergistic effects. Additional studies
both in humans and animals will be required to identify precisely
which components and constituents have beneficial roles. Unfortu-
nately, comparisons between different animal or clinical studies are
hampered by the lack of standardization of soy nomenclature, the
different formulations, doses, routes of exposure, time and duration
of exposure as well as by major differences in the subsequent analy-
ses performed to evaluate the effects and elucidate the mechanisms
by which phytoestrogens and soy potentially improve glucose and
lipid metabolism. All of these variables make it difficult to compare
and evaluate the putative beneficial effects of soy and phytoestro-
gens on metabolism. Clearly more standardized studies, involving
both basic research and clinical trials, are needed. Given the rapidly
increasing prevalence and societal impact of metabolic disorders,
such studies should have high priority.
Acknowledgments
We thank Prof. J.-D. Vassalli for critical comments on the
manuscript. Authors were funded by grants from the Swiss National
Science Foundation, Foundation Gertrude von Meissner, Fondation
Ernst & Lucie Schmidheiny, the Sir Jules Thorn Charitable Over-
seas Trust Reg., Schaan and the Cloëtta foundation. Serge Nef is a
founder of Amazentis S.A. and a member of its scientific advisory
board.
References
Adachi, T., Yasuda, K., Mori, C., Yashinaga, M., Aoki, N., Tsujimoto, G., Tsuda, K.,
2005. Promoting insulin secretion in pancreatic islets by means of bisphenol-
A and nonylphenol via intracellular estrogen-receptors. Food Chem. Toxicol. 43,
713–719.
Adlercreutz, H., Fotsis, T., Lampe, J., Wahala, K., Makela, T., Brunow, G., Hase, T., 1993a.
Quantitative determination of lignans and isoflavonoids in plasma of omniv-
orous and vegetarian women by isotope dilution gas chromatography–mass
spectrometry. Scand. J. Clin. Lab. Invest. 215 (Suppl.), 5–18.
Adlercreutz, H., Markkanen, H., Watanabe, S., 1993b. Plasma concentrations of phyto-
oestrogens in Japanese men. Lancet 342, 1209–1210.
Ae Park, S., Choi, M.S., Cho, S.Y., Seo, J.S., Jung, U.J., Kim, M.J., Sung, M.K., Park, Y.B., Lee,
M.K., 2006. Genistein and daidzein modulate hepatic glucose and lipid regulating
enzyme activities in C57BL/KsJ-db/db mice. Life Sci. 79, 1207–1213.
Ahmed-Sorour, H., Bailey, C.J., 1980. Role of ovarian hormones in the long-term con-
trol of glucose homeostasis. Interaction with insulin, glucagon and epinephrine.
Horm. Res. 13, 396–403.
Akiyama, T., Ishida, J., Nakagawa, S., Ogawara, H., Watanabe, S., Itoh, N., Shibuya,
M., Fukami, Y., 1987. Genistein, a specific inhibitor of tyrosine-specific protein
kinases. J. Biol. Chem. 262, 5592–5595.
Ali, A.A., Velasquez, M.T., Hansen, C.T., Mohamed, A.I., Bhathena, S.J., 2005. Modula-
tion of carbohydrate metabolism and peptide hormones by soybean isoflavones
and probiotics in obesity and diabetes. J. Nutr. Biochem. 16, 693–699.
Allison, D.B., Gadbury, G., Schwartz, L.G., Murugesan, R., Kraker, J.L., Heshka, S.,
Fontaine, K.R., Heymsfield, S.B., 2003. A novel soy-based meal replacement for-
mula for weight loss among obese individuals: a randomized controlled clinical
trial. Eur. J. Clin. Nutr. 57, 514–522.
Alonso-Magdalena, P., Morimoto, S., Ripoll, C., Fuentes, E., Nadal, A., 2006. The estro-
genic effect of bisphenol A disrupts pancreatic beta-cell function in vivo and
induces insulin resistance. Environ. Health Perspect. 114, 106–112.
Alonso-Magdalena, P., Ropero, A.B., Carrera, M.P., Cederroth, C.R., Baquie, M., Gau-
thier, B.R., Nef, S., Stefani, E., Nadal, A., 2008. Pancreatic insulin content regulation
by the estrogen receptor ER alpha. PLoS ONE 3, e2069.
Anderson, J.W., Johnstone, B.M., Cook-Newell, M.E., 1995. Meta-analysis of the effects
of soy protein intake on serum lipids. N. Engl. J. Med. 333, 276–282.
Anderson, J.W., Hoie, L.H., 2005. Weight loss and lipid changes with low-energy
diets: comparator study of milk-based versus soy-based liquid meal replacement
interventions. J. Am. Coll. Nutr. 24, 210–216.
Anderson, J.W., Fuller, J., Patterson, K., Blair, R., Tabor, A., 2007. Soy compared to
casein meal replacement shakes with energy-restricted diets for obese women:
randomized controlled trial. Metabolism 56, 280–288.
Aoyama, T., Fukui, K., Nakamori, T., Hashimoto, Y., Yamamoto, T., Takamatsu, K.,
Sugano, M., 2000a. Effect of soy and milk whey protein isolates and their
hydrolysates on weight reduction in genetically obese mice. Biosci. Biotechnol.
Biochem. 64, 2594–2600.
Aoyama, T., Fukui, K., Takamatsu, K., Hashimoto, Y., Yamamoto, T., 2000b. Soy protein
isolate and its hydrolysate reduce body fat of dietary obese rats and genetically
obese mice (yellow KK). Nutrition 16, 349–354.
Atkinson, C., Frankenfeld, C.L., Lampe, J.W., 2005. Gut bacterial metabolism of the soy
isoflavone daidzein: exploring the relevance to human health. Exp. Biol. Med.
(Maywood) 230, 155–170.
Bailey, C.J., Matty, A.J., 1972. Glucose tolerance and plasma insulin of the rat in relation
to the oestrous cycle and sex hormones. Horm. Metab. Res. 4, 266–270.
Bailey, C.J., Ahmed-Sorour, H., 1980. Role of ovarian hormones in the long-term
control of glucose homeostasis. Effects of insulin secretion. Diabetologia 19,
475–481.
Barros, R.P., Machado, U.F., Warner, M., Gustafsson, J.A., 2006. Muscle GLUT4 regula-
tion by estrogen receptors ERbeta and ERalpha. Proc. Natl. Acad. Sci. U.S.A. 103,
1605–1608.
Bates, S.H., Myers Jr., M.G., 2003. The role of leptin receptor signaling in feeding and
neuroendocrine function. Trends Endocrinol. Metab. 14, 447–452.
Bennetts, H.W., Underwood, E.J., Shier, F.L., 1946. A specific breeding problem of
sheep on subterranean clover pastures in Western Australia. Austr. Vet. J. 22,
2–12.
Bhathena, S.J., Velasquez, M.T., 2002. Beneficial role of dietary phytoestrogens in
obesity and diabetes. Am. J. Clin. Nutr. 76, 1191–1201.
Bu, L., Setchell, K.D., Lephart, E.D., 2005. Influences of dietary soy isoflavones on
metabolism but not nociception and stress hormone responses in ovariec-
tomized female rats. Reprod. Biol. Endocrinol. 3, 58.
Bu, L.H., Lephart, E.D., 2005. Effects of dietary phytoestrogens on core body temper-
ature during the estrous cycle and pregnancy. Brain Res. Bull. 65, 219–223.
Caldwell, C.R., Britz, S.J., Mirecki, R.M., 2005. Effect of temperature, elevated car-
bon dioxide, and drought during seed development on the isoflavone content
of dwarf soybean [Glycine max (L.) Merrill] grown in controlled environments. J.
Agric. Food Chem. 53, 1125–1129.
Carani, C., Qin, K., Simoni, M., Faustini-Fustini, M., Serpente, S., Boyd, J., Korach, K.S.,
Simpson, E.R., 1997. Effect of testosterone and estradiol in a man with aromatase
deficiency. N. Engl. J. Med. 337, 91–95.
Case, A.M., Reid, R.L., 2001. Menstrual cycle effects on common medical conditions.
Compr. Ther. 27, 65–71.
Cederroth, C.R., Vinciguerra, M., Kuhne, F., Madani, R., Doerge, D.R., Visser, T.J., Foti,
M., Rohner-Jeanrenaud, F., Vassalli, J.D., Nef, S., 2007. A phytoestrogen-rich diet
increases energy expenditure and decreases adiposity in mice. Environ. Health
Perspect. 115, 1467–1473.
Cederroth, C.R., Vinciguerra, M., Gjinovci, A., Kuhne, F., Klein, M., Cederroth, M., Caille,
D., Suter, M., Neumann, D., James, R.W., Doerge, D.R., Wallimann, T., Meda, P., Foti,
M., Rohner-Jeanrenaud, F., Vassalli, J.D., Nef, S., 2008. Dietary phytoestrogens
activate AMP-activated protein kinase with improvement in lipid and glucose
metabolism. Diabetes 57, 1176–1185.
Chen, A.C., Donovan, S.M., 2004. Genistein at a concentration present in soy infant
formula inhibits Caco-2BBe cell proliferation by causing G2/M cell cycle arrest.
J. Nutr. 134, 1303–1308.
Cheng, S.Y., Shaw, N.S., Tsai, K.S., Chen, C.Y., 2004. The hypoglycemic effects of
soy isoflavones on postmenopausal women. J. Women Health (Larchmt) 13,
1080–1086.
Choi, M.S., Jung, U.J., Yeo, J., Kim, M.J., Lee, M.K., 2008. Genistein and daidzein prevent
diabetes onset by elevating insulin level and altering hepatic gluconeogenic and
lipogenic enzyme activities in non-obese diabetic (NOD) mice. Diabetes Metab.
Res. Rev. 24, 74–81.
Cooke, P.S., Heine, P.A., Taylor, J.A., Lubahn, D.B., 2001. The role of estrogen and
estrogen receptor-alpha in male adipose tissue. Mol. Cell. Endocrinol. 178, 147–
154.
Cooke, P.S., Naaz, A., 2004. Role of estrogens in adipocyte development and function.
Exp. Biol. Med. (Maywood) 229, 1127–1135.
Coward, L., Kirk, M., Albin, N., Barnes, S., 1996. Analysis of plasma isoflavones
by reversed-phase HPLC-multiple reaction ion monitoring-mass spectrometry.
Clin. Chim. Acta 247, 121–142.
Crouse 3rd, J.R., Morgan, T., Terry, J.G., Ellis, J., Vitolins, M., Burke, G.L., 1999. A ran-
domized trial comparing the effect of casein with that of soy protein containing
varying amounts of isoflavones on plasma concentrations of lipids and lipopro-
teins. Arch. Intern. Med. 159, 2070–2076.
Dalais, F.S., Ebeling, P.R., Kotsopoulos, D., McGrath, B.P., Teede, H.J., 2003. The effects
of soy protein containing isoflavones on lipids and indices of bone resorption in
postmenopausal women. Clin. Endocrinol. (Oxf.) 58, 704–709.
Davis, J., Iqbal, M.J., Steinle, J., Oitker, J., Higginbotham, D.A., Peterson, R.G., Banz,
W.J., 2005. Soy protein influences the development of the metabolic syndrome
in male obese ZDFxSHHF rats. Horm. Metab. Res. 37, 316–325.
de Kleijn, M.J., van der Schouw, Y.T., Wilson, P.W., Grobbee, D.E., Jacques, P.F., 2002.
Dietary intake of phytoestrogens is associated with a favorable metabolic car-
diovascular risk profile in postmenopausal U.S. women: the Framingham study.
J. Nutr. 132, 276–282.
Dixon, R.A., 2004. Phytoestrogens. Annu. Rev. Plant Biol. 55, 225–261.
Dolinoy, D.C., Weidman, J.R., Waterland, R.A., Jirtle, R.L., 2006. Maternal genistein
alters coat color and protects Avy mouse offspring from obesity by modifying
the fetal epigenome. Environ. Health Perspect. 114, 567–572.
Duffy, C., Perez, K., Partridge, A., 2007. Implications of phytoestrogen intake for breast
cancer. CA Cancer J. Clin. 57, 260–277.
Engelgau, M.M., Geiss, L.S., Saaddine, J.B., Boyle, J.P., Benjamin, S.M., Gregg, E.W.,
Tierney, E.F., Rios-Burrows, N., Mokdad, A.H., Ford, E.S., Imperatore, G., Narayan,
 C.R. Cederroth, S. Nef / Molecular and Cellular Endocrinology 304 (2009) 30–42
K.M., 2004. The evolving diabetes burden in the United States. Ann. Intern. Med.
140, 945–950.
Foryst-Ludwig, A., Clemenz, M., Hohmann, S., Hartge, M., Sprang, C., Frost, N., Krikov,
M., Bhanot, S., Barros, R., Morani, A., Gustafsson, J.A., Unger, T., Kintscher, U., 2008.
Metabolic actions of estrogen receptor beta (ERbeta) are mediated by a negative
cross-talk with PPARgamma. PLoS Genet. 4, e1000108.
Fujimoto, W.Y., Leonetti, D.L., Kinyoun, J.L., Newell-Morris, L., Shuman, W.P., Stolov,
W.C., Wahl, P.W., 1987. Prevalence of diabetes mellitus and impaired glu-
cose tolerance among second-generation Japanese-American men. Diabetes 36,
721–729.
Fujimoto, W.Y., Leonetti, D.L., Bergstrom, R.W., Kinyoun, J.L., Stolov, W.C., Wahl, P.W.,
1991. Glucose intolerance and diabetic complications among Japanese-American
women. Diabetes Res. Clin. Pract. 13, 119–129.
Gambacciani, M., Ciaponi, M., Cappagli, B., Piaggesi, L., De Simone, L., Orlandi, R.,
Genazzani, A.R., 1997. Body weight, body fat distribution, and hormonal replace-
ment therapy in early postmenopausal women. J. Clin. Endocrinol. Metab. 82,
414–417.
Gao, Q., Mezei, G., Nie, Y., Rao, Y., Choi, C.S., Bechmann, I., Leranth, C., Toran-Allerand,
D., Priest, C.A., Roberts, J.L., Gao, X.B., Mobbs, C., Shulman, G.I., Diano, S., Hor-
vath, T.L., 2007. Anorectic estrogen mimics leptin’s effect on the rewiring of
melanocortin cells and Stat3 signaling in obese animals. Nat. Med. 13, 89–94.
Garcia, M.C., Torre, M., Marina, M.L., Laborda, F., 1997. Composition and characteri-
zation of soyabean and related products. Crit. Rev. Food Sci. Nutr. 37, 361–391.
Gardner, C.D., Newell, K.A., Cherin, R., Haskell, W.L., 2001. The effect of soy pro-
tein with or without isoflavones relative to milk protein on plasma lipids
in hypercholesterolemic postmenopausal women. Am. J. Clin. Nutr. 73, 728–
735.
Godsland, I.F., 2005. Oestrogens and insulin secretion. Diabetologia 48, 2213–2220.
Goodman-Gruen, D., Kritz-Silverstein, D., 2001. Usual dietary isoflavone intake is
associated with cardiovascular disease risk factors in postmenopausal women.
J. Nutr. 131, 1202–1206.
Goodman-Gruen, D., Kritz-Silverstein, D., 2003. Usual dietary isoflavone intake and
body composition in postmenopausal women. Menopause 10, 427–432.
Greany, K.A., Nettleton, J.A., Wangen, K.E., Thomas, W., Kurzer, M.S., 2004. Probi-
otic consumption does not enhance the cholesterol-lowering effect of soy in
postmenopausal women. J. Nutr. 134, 3277–3283.
Grieshop, C.M., Fahey Jr., G.C., 2001. Comparison of quality characteristics of soybeans
from Brazil, China, and the United States. J. Agric. Food Chem. 49, 2669–2673.
Grieshop, C.M., Kadzere, C.T., Clapper, G.M., Flickinger, E.A., Bauer, L.L., Frazier, R.L.,
Fahey Jr., G.C., 2003. Chemical and nutritional characteristics of United States
soybeans and soybean meals. J. Agric. Food Chem. 51, 7684–7691.
Guthrie, J.R., Ball, M., Murkies, A., Dennerstein, L., 2000. Dietary phytoestrogen intake
in mid-life Australian-born women: relationship to health variables. Climacteric
3, 254–261.
Hall, W.L., Vafeiadou, K., Hallund, J., Bugel, S., Reimann, M., Koebnick, C., Zunft,
H.J., Ferrari, M., Branca, F., Dadd, T., Talbot, D., Powell, J., Minihane, A.M., Cas-
sidy, A., Nilsson, M., Dahlman-Wright, K., Gustafsson, J.A., Williams, C.M., 2006.
Soy-isoflavone-enriched foods and markers of lipid and glucose metabolism in
postmenopausal women: interactions with genotype and equol production. Am.
J. Clin. Nutr. 83, 592–600.
Heine, P.A., Taylor, J.A., Iwamoto, G.A., Lubahn, D.B., Cooke, P.S., 2000. Increased adi-
pose tissue in male and female estrogen receptor-alpha knockout mice. Proc.
Natl. Acad. Sci. U.S.A. 97, 12729–12734.
Howitz, K.T., Sinclair, D.A., 2008. Xenohormesis: sensing the chemical cues of other
species. Cell 133, 387–391.
Hurley, C., Richard, D., Deshaies, Y., Jacques, H., 1998. Soy protein isolate in the pres-
ence of cornstarch reduces body fat gain in rats. Can. J. Physiol. Pharmacol. 76,
1000–1007.
Hwang, C.S., Kwak, H.S., Lim, H.J., Lee, S.H., Kang, Y.S., Choe, T.B., Hur, H.G., Han, K.O.,
2006. Isoflavone metabolites and their in vitro dual functions: they can act as
an estrogenic agonist or antagonist depending on the estrogen concentration. J.
Steroid Biochem. Mol. Biol. 101, 246–253.
Hwang, J.T., Park, I.J., Shin, J.I., Lee, Y.K., Lee, S.K., Baik, H.W., Ha, J., Park, O.J., 2005.
Genistein, EGCG, and capsaicin inhibit adipocyte differentiation process via acti-
vating AMP-activated protein kinase. Biochem. Biophys. Res. Commun. 338,
694–699.
Ikeda, Y., Iki, M., Morita, A., Kajita, E., Kagamimori, S., Kagawa, Y., Yoneshima, H., 2006.
Intake of fermented soybeans, natto, is associated with reduced bone loss in
postmenopausal women: Japanese Population-Based Osteoporosis (JPOS) Study.
J. Nutr. 136, 1323–1328.
Jang, E.H., Moon, J.S., Ko, J.H., Ahn, C.W., Lee, H.H., Shin, J.K., Park, C.S., Kang, J.H.,
2008. Novel black soy peptides with antiobesity effects: activation of leptin-like
signaling and AMP-activated protein kinase. Int. J. Obes. (Lond.) 32, 1161–1170.
Jayagopal, V., Albertazzi, P., Kilpatrick, E.S., Howarth, E.M., Jennings, P.E., Hepburn,
D.A., Atkin, S.L., 2002. Beneficial effects of soy phytoestrogen intake in post-
menopausal women with type 2 diabetes. Diabetes Care 25, 1709–1714.
Jefferson, W.N., Padilla-Banks, E., Newbold, R.R., 2005. Adverse effects on female
development and reproduction in CD-1 mice following neonatal exposure to
the phytoestrogen genistein at environmentally relevant doses. Biol. Reprod. 73,
798–806.
Jefferson, W.N., Padilla-Banks, E., Newbold, R.R., 2007. Disruption of the female repro-
ductive system by the phytoestrogen genistein. Reprod. Toxicol. 23, 308–316.
Jones, M.E., Thorburn, A.W., Britt, K.L., Hewitt, K.N., Wreford, N.G., Proietto, J., Oz,
O.K., Leury, B.J., Robertson, K.M., Yao, S., Simpson, E.R., 2000. Aromatase-deficient
(ArKO) mice have a phenotype of increased adiposity. Proc. Natl. Acad. Sci. U.S.A.
97, 12735–12740.
41
Ju, Y.H., Doerge, D.R., Allred, K.F., Allred, C.D., Helferich, W.G., 2002. Dietary genis-
tein negates the inhibitory effect of tamoxifen on growth of estrogen-dependent
human breast cancer (MCF-7) cells implanted in athymic mice. Cancer Res. 62,
2474–2477.
Kim, E.K., Kwon, K.B., Song, M.Y., Seo, S.W., Park, S.J., Ka, S.O., Na, L., Kim, K.A., Ryu,
D.G., So, H.S., Park, R., Park, J.W., Park, B.H., 2007. Genistein protects pancreatic
beta cells against cytokine-mediated toxicity. Mol. Cell. Endocrinol. 278, 18–28.
King, R.A., Bursill, D.B., 1998. Plasma and urinary kinetics of the isoflavones daidzein
and genistein after a single soy meal in humans. Am. J. Clin. Nutr. 67, 867–872.
Kirk, E.A., Sutherland, P., Wang, S.A., Chait, A., LeBoeuf, R.C., 1998. Dietary isoflavones
reduce plasma cholesterol and atherosclerosis in C57BL/6 mice but not LDL
receptor-deficient mice. J. Nutr. 128, 954–959.
Kohno, M., Hirotsuka, M., Kito, M., Matsuzawa, Y., 2006. Decreases in serum tri-
acylglycerol and visceral fat mediated by dietary soybean beta-conglycinin. J.
Atheroscler. Thromb. 13, 247–255.
Kok, L., Kreijkamp-Kaspers, S., Grobbee, D.E., Lampe, J.W., van der Schouw, Y.T., 2005.
Soy isoflavones, body composition, and physical performance. Maturitas 52,
102–110.
Kuiper, G.G., Carlsson, B., Grandien, K., Enmark, E., Haggblad, J., Nilsson, S., Gustafs-
son, J.A., 1997. Comparison of the ligand binding specificity and transcript tissue
distribution of estrogen receptors alpha and beta. Endocrinology 138, 863–
870.
Kuiper, G.G., Lemmen, J.G., Carlsson, B., Corton, J.C., Safe, S.H., van der Saag, P.T.,
van der Burg, B., Gustafsson, J.A., 1998. Interaction of estrogenic chemicals and
phytoestrogens with estrogen receptor beta. Endocrinology 139, 4252–4263.
Lavigne, C., Marette, A., Jacques, H., 2000. Cod and soy proteins compared with
casein improve glucose tolerance and insulin sensitivity in rats. Am. J. Physiol.
Endocrinol. Metab. 278, E491–500.
Le May, C., Chu, K., Hu, M., Ortega, C.S., Simpson, E.R., Korach, K.S., Tsai, M.J.,
Mauvais-Jarvis, F., 2006. Estrogens protect pancreatic beta-cells from apopto-
sis and prevent insulin-deficient diabetes mellitus in mice. Proc. Natl. Acad. Sci.
U.S.A. 103, 9232–9237.
Lee, J.S., 2006. Effects of soy protein and genistein on blood glucose, antioxidant
enzyme activities, and lipid profile in streptozotocin-induced diabetic rats. Life
Sci. 79, 1578–1584.
Leopold, A.S., Erwin, M., Oh, J., Browning, B., 1976. Phytoestrogens: adverse effects
on reproduction in California quail. Science 191, 98–100.
Lephart, E.D., Rhees, R.W., Setchell, K.D., Bu, L.H., Lund, T.D., 2003. Estrogens and
phytoestrogens: brain plasticity of sexually dimorphic brain volumes. J. Steroid
Biochem. Mol. Biol. 85, 299–309.
Lephart, E.D., Porter, J.P., Lund, T.D., Bu, L., Setchell, K.D., Ramoz, G., Crowley, W.R.,
2004a. Dietary isoflavones alter regulatory behaviors, metabolic hormones and
neuroendocrine function in Long-Evans male rats. Nutr. Metab. (Lond.) 1, 16.
Lephart, E.D., Setchell, K.D., Handa, R.J., Lund, T.D., 2004b. Behavioral effects of
endocrine-disrupting substances: phytoestrogens. Ilar. J. 45, 443–454.
Li, Z., Hong, K., Saltsman, P., DeShields, S., Bellman, M., Thames, G., Liu, Y., Wang, H.J.,
Elashoff, R., Heber, D., 2005. Long-term efficacy of soy-based meal replacements
vs an individualized diet plan in obese type II DM patients: relative effects on
weight loss, metabolic parameters, and C-reactive protein. Eur. J. Clin. Nutr. 59,
411–418.
Liang, Y.Q., Akishita, M., Kim, S., Ako, J., Hashimoto, M., Iijima, K., Ohike, Y., Watanabe,
T., Sudoh, N., Toba, K., Yoshizumi, M., Ouchi, Y., 2002. Estrogen receptor beta is
involved in the anorectic action of estrogen. Int. J. Obes. Relat. Metab. Disord. 26,
1103–1109.
Lin, J., Handschin, C., Spiegelman, B.M., 2005. Metabolic control through the PGC-1
family of transcription coactivators. Cell. Metab. 1, 361–370.
Liu, B., Edgerton, S., Yang, X., Kim, A., Ordonez-Ercan, D., Mason, T., Alvarez, K.,
McKimmey, C., Liu, N., Thor, A., 2005. Low-dose dietary phytoestrogen abrogates
tamoxifen-associated mammary tumor prevention. Cancer Res. 65, 879–886.
Liu, D., Zhen, W., Yang, Z., Carter, J.D., Si, H., Reynolds, K.A., 2006. Genistein acutely
stimulates insulin secretion in pancreatic beta-cells through a cAMP-dependent
protein kinase pathway. Diabetes 55, 1043–1050.
Louet, J.F., LeMay, C., Mauvais-Jarvis, F., 2004. Antidiabetic actions of estrogen: insight
from human and genetic mouse models. Curr. Atheroscler. Rep. 6, 180–185.
Lukaczer, D., Liska, D.J., Lerman, R.H., Darland, G., Schiltz, B., Tripp, M., Bland, J.S.,
2006. Effect of a low glycemic index diet with soy protein and phytosterols on
CVD risk factors in postmenopausal women. Nutrition 22, 104–113.
Makela, S., Davis, V.L., Tally, W.C., Korkman, J., Salo, L., Vihko, R., Santti, R., Korach,
K.S., 1994. Dietary estrogens act through estrogen receptor-mediated processes
and show no antiestrogenicity in cultured breast cancer cells. Environ. Health
Perspect. 102, 572–578.
Makela, S., Santti, R., Salo, L., McLachlan, J.A., 1995. Phytoestrogens are partial estro-
gen agonists in the adult male mouse. Environ. Health Perspect. 103 (Suppl 7),
123–127.
Makimura, H., Mizuno, T.M., Mastaitis, J.W., Agami, R., Mobbs, C.V., 2002. Reducing
hypothalamic AGRP by RNA interference increases metabolic rate and decreases
body weight without influencing food intake. BMC Neurosci. 3, 18.
Martensson, U.E.A., Salehi, A.S., Windahl, S., Gomez, M.F., Swärd, K., Daszkiewicz-
Nilsson, J., Wendt, A., Andersson, N., Hellstrand, P., Grände, P.-O., Owman, C.,
Rosen, C.J., Adamo, M.L., Lundquist, I., Rorsman, P., Nilsson, B.-O., Ohlsson, C.,
Olde, B., Leeb-Lundberg, L.M.F., 2008. Deletion of the G protein-coupled recep-
tor GRP30 impairs glucose tolerance, reduces bone growth, increases blood
pressure, and eliminates estradiol-stimulated insulin release in female mice.
Endocrinology, 2008–2623, 10. 1210/en.
Meli, R., Pacilio, M., Raso, G.M., Esposito, E., Coppola, A., Nasti, A., Di Carlo, C., Nappi,
C., Di Carlo, R., 2004. Estrogen and raloxifene modulate leptin and its receptor in
42 C.R. Cederroth, S. Nef / Molecular and Cellular Endocrinology 304 (2009) 30–42
hypothalamus and adipose tissue from ovariectomized rats. Endocrinology 145,
3115–3121.
Moriyama, T., Kishimoto, K., Nagai, K., Urade, R., Ogawa, T., Utsumi, S., Maruyama, N.,
Maebuchi, M., 2004. Soybean beta-conglycinin diet suppresses serum triglyc-
eride levels in normal and genetically obese mice by induction of beta-oxidation,
downregulation of fatty acid synthase, and inhibition of triglyceride absorption.
Biosci. Biotechnol. Biochem. 68, 352–359.
Morton, M.S., Wilcox, G., Wahlqvist, M.L., Griffiths, K., 1994. Determination of lignans
and isoflavonoids in human female plasma following dietary supplementation.
J. Endocrinol. 142, 251–259.
Musatov, S., Chen, W., Pfaff, D.W., Mobbs, C.V., Yang, X.J., Clegg, D.J., Kaplitt, M.G.,
Ogawa, S., 2007. Silencing of estrogen receptor alpha in the ventromedial nucleus
of hypothalamus leads to metabolic syndrome. Proc. Natl. Acad. Sci. U.S.A. 104,
2501–2506.
Nadal, A., Rovira, J.M., Laribi, O., Leon-Quinto, T., Andreu, E., Ripoll, C., Soria, B., 1998.
Rapid insulinotropic effect of 17beta-estradiol via a plasma membrane receptor.
FASEB J. 12, 1341–1348.
Naaz, A., Yellayi, S., Zakroczymski, M.A., Bunick, D., Doerge, D.R., Lubahn, D.B.,
Helferich, W.G., Cooke, P.S., 2003. The soy isoflavone genistein decreases adipose
deposition in mice. Endocrinology 144, 3315–3320.
Nagasawa, A., Fukui, K., Kojima, M., Kishida, K., Maeda, N., Nagaretani, H., Hibuse, T.,
Nishizawa, H., Kihara, S., Waki, M., Takamatsu, K., Funahashi, T., Matsuzawa, Y.,
2003. Divergent effects of soy protein diet on the expression of adipocytokines.
Biochem. Biophys. Res. Commun. 311, 909–914.
Nemoto, Y., Toda, K., Ono, M., Fujikawa-Adachi, K., Saibara, T., Onishi, S., Enzan,
H., Okada, T., Shizuta, Y., 2000. Altered expression of fatty acid-metabolizing
enzymes in aromatase-deficient mice. J. Clin. Invest. 105, 1819–1825.
Newbold, R.R., Banks, E.P., Bullock, B., Jefferson, W.N., 2001. Uterine adenocarcinoma
in mice treated neonatally with genistein. Cancer Res. 61, 4325–4328.
Nordentoft, I., Jeppesen, P.B., Hong, J., Abudula, R., Hermansen, K., 2008. Increased
insulin sensitivity and changes in the expression profile of key insulin regulatory
genes and beta cell transcription factors in diabetic KKAy-mice after feeding with
a soy bean protein rich diet high in isoflavone content. J. Agric. Food Chem. 56,
4377–4385.
Oakenfull, D., 2001. Soy protein, saponins and plasma cholesterol. J. Nutr. 131,
2971–2972.
Ogawa, S., Chan, J., Gustafsson, J.A., Korach, K.S., Pfaff, D.W., 2003. Estrogen increases
locomotor activity in mice through estrogen receptor alpha: specificity for the
type of activity. Endocrinology 144, 230–239.
Ohlsson, C., Hellberg, N., Parini, P., Vidal, O., Bohlooly, Y.M., Rudling, M., Lindberg,
M.K., Warner, M., Angelin, B., Gustafsson, J.A., 2000. Obesity and disturbed
lipoprotein profile in estrogen receptor-alpha-deficient male mice. Biochem.
Biophys. Res. Commun. 278, 640–645.
Olshansky, S.J., Passaro, D.J., Hershow, R.C., Layden, J., Carnes, B.A., Brody, J., Hayflick,
L., Butler, R.N., Allison, D.B., Ludwig, D.S., 2005. A potential decline in life
expectancy in the United States in the 21st century. N. Engl. J. Med. 352,
1138–1145.
Penza, M., Montani, C., Romani, A., Vignolini, P., Pampaloni, B., Tanini, A., Brandi, M.L.,
Alonso-Magdalena, P., Nadal, A., Ottobrini, L., Parolini, O., Bignotti, E., Calza, S.,
Maggi, A., Grigolato, P.G., Di Lorenzo, D., 2006. Genistein affects adipose tissue
deposition in a dose-dependent and gender-specific manner. Endocrinology 147,
5740–5751.
Ruderman, N., Prentki, M., 2004. AMP kinase and malonyl-CoA: targets for therapy
of the metabolic syndrome. Nat. Rev. Drug. Discov. 3, 340–351.
Ruhlen, R.L., Howdeshell, K.L., Mao, J., Taylor, J.A., Bronson, F.H., Newbold, R.R.,
Welshons, W.V., vom Saal, F.S., 2008. Low phytoestrogen levels in feed increase
fetal serum estradiol resulting in the “fetal estrogenization syndrome” and obe-
sity in CD-1 mice. Environ. Health Perspect. 116, 322–328.
Sacks, F.M., Lichtenstein, A., Van Horn, L., Harris, W., Kris-Etherton, P., Winston, M.,
2006. Soy protein, isoflavones, and cardiovascular health: an American Heart
Association Science Advisory for professionals from the Nutrition Committee.
Circulation 113, 1034–1044.
Sanders, T.A., Dean, T.S., Grainger, D., Miller, G.J., Wiseman, H., 2002. Moderate
intakes of intact soy protein rich in isoflavones compared with ethanol-extracted
soy protein increase HDL but do not influence transforming growth factor beta(1)
concentrations and hemostatic risk factors for coronary heart disease in healthy
subjects. Am. J. Clin. Nutr. 76, 373–377.
Schwartz, M.W., Woods, S.C., Porte Jr., D., Seeley, R.J., Baskin, D.G., 2000. Central
nervous system control of food intake. Nature 404, 661–671.
Setchell, K.D., 1998. Phytoestrogens: the biochemistry, physiology, and implications
for human health of soy isoflavones. Am. J. Clin. Nutr. 68, 1333S–1346S.
Setchell, K.D., Brown, N.M., Desai, P., Zimmer-Nechemias, L., Wolfe, B.E., Bras-
hear, W.T., Kirschner, A.S., Cassidy, A., Heubi, J.E., 2001. Bioavailability of pure
isoflavones in healthy humans and analysis of commercial soy isoflavone sup-
plements. J. Nutr. 131, S1362–S1375.
Setchell, K.D., 2006. Assessing risks and benefits of genistein and soy. Environ. Health
Perspect. 114, A332–A333.
Stutz, A.M., Morrison, C.D., Argyropoulos, G., 2005. The Agouti-related protein and
its role in energy homeostasis. Peptides 26, 1771–1781.
Szkudelska, K., Nogowski, L., Szkudelski, T., 2000. Genistein affects lipogenesis and
lipolysis in isolated rat adipocytes. J. Steroid Biochem. Mol. Biol. 75, 265–271.
Takatsuka, N., Nagata, C., Kurisu, Y., Inaba, S., Kawakami, N., Shimizu, H., 2000.
Hypocholesterolemic effect of soymilk supplementation with usual diet
in premenopausal normolipidemic Japanese women. Prev. Med. 31, 308–
314.
Takeda, K., Toda, K., Saibara, T., Nakagawa, M., Saika, K., Onishi, T., Sugiura, T., Shizuta,
Y., 2003. Progressive development of insulin resistance phenotype in male mice
with complete aromatase (CYP19) deficiency. J. Endocrinol. 176, 237–246.
Teixeira, S.R., Potter, S.M., Weigel, R., Hannum, S., Erdman Jr., J.W., Hasler, C.M., 2000.
Effects of feeding 4 levels of soy protein for 3 and 6 wk on blood lipids and
apolipoproteins in moderately hypercholesterolemic men. Am. J. Clin. Nutr. 71,
1077–1084.
Tham, D.M., Gardner, C.D., Haskell, W.L., 1998. Clinical review 97: potential health
benefits of dietary phytoestrogens: a review of the clinical, epidemiological, and
mechanistic evidence. J. Clin. Endocrinol. Metab. 83, 2223–2235.
Thigpen, J.E., Haseman, J.K., Saunders, H.E., Setchell, K.D., Grant, M.G., Forsythe, D.B.,
2003. Dietary phytoestrogens accelerate the time of vaginal opening in imma-
ture CD-1 mice. Comp. Med. 53, 607–615.
Torre-Villalvazo, I., Tovar, A.R., Ramos-Barragan, V.E., Cerbon-Cervantes, M.A., Torres,
N., 2008. Soy protein ameliorates metabolic abnormalities in liver and adipose
tissue of rats fed a high fat diet. J. Nutr. 138, 462–468.
United States Department of Agriculture, 1979. Composition of foods: fats and oils.
In: USDA Handbook 8-4. USDA, Washington, DC.
van Erp-Baart, M.A., Brants, H.A., Kiely, M., Mulligan, A., Turrini, A., Sermoneta, C.,
Kilkkinen, A., Valsta, L.M., 2003. Isoflavone intake in four different European
countries: the VENUS approach. Br. J. Nutr. 89 (Suppl 1), S25–S30.
Vedavanam, K., Srijayanta, S., O’Reilly, J., Raman, A., Wiseman, H., 1999. Antioxi-
dant action and potential antidiabetic properties of an isoflavonoid-containing
soyabean phytochemical extract (SPE). Phytother. Res. 13, 601–608.
Velasquez, M.T., Bhathena, S.J., 2007. Role of dietary soy protein in obesity. Int. J. Med.
Sci. 4, 72–82.
Wang, H.J., Murphy, P.A., 1994. Isoflavone composition of American and Japanese
soybeans in Iowa: effects of variety, crop year, and location. J. Agric. Food Chem.
42, 1674–1677.
Watanabe, S., Yamaguchi, M., Sobue, T., Takahashi, T., Miura, T., Arai, Y., Mazur, W.,
Wahala, K., Adlercreutz, H., 1998. Pharmacokinetics of soybean isoflavones in
plasma, urine and feces of men after ingestion of 60 g baked soybean powder
(kinako). J. Nutr. 128, 1710–1715.
Weber, K.S., Setchell, K.D., Stocco, D.M., Lephart, E.D., 2001. Dietary soy-
phytoestrogens decrease testosterone levels and prostate weight without
altering LH, prostate 5alpha-reductase or testicular steroidogenic acute reg-
ulatory peptide levels in adult male Sprague–Dawley rats. J. Endocrinol. 170,
591–599.
Weickert, M.O., Reimann, M., Otto, B., Hall, W.L., Vafeiadou, K., Hallund, J., Ferrari, M.,
Talbot, D., Branca, F., Bugel, S., Williams, C.M., Zunft, H.J., Koebnick, C., 2006. Soy
isoflavones increase preprandial peptide YY (PYY), but have no effect on ghrelin
and body weight in healthy postmenopausal women. J. Negat. Results. Biomed.
5, 11.
Winder, W.W., Hardie, D.G., 1996. Inactivation of acetyl-CoA carboxylase and activa-
tion of AMP-activated protein kinase in muscle during exercise. Am. J. Physiol.
270, E299–304.
Wortley, K.E., Anderson, K.D., Yasenchak, J., Murphy, A., Valenzuela, D., Diano, S.,
Yancopoulos, G.D., Wiegand, S.J., Sleeman, M.W., 2005. Agouti-related protein-
deficient mice display an age-related lean phenotype. Cell. Metab. 2, 421–
427.
Wu, A.H., Wan, P., Hankin, J., Tseng, C.C., Yu, M.C., Pike, M.C., 2002. Adolescent and
adult soy intake and risk of breast cancer in Asian-Americans. Carcinogenesis
23, 1491–1496.
Wu, J., Wang, X., Chiba, H., Higuchi, M., Nakatani, T., Ezaki, O., Cui, H., Yamada, K.,
Ishimi, Y., 2004. Combined intervention of soy isoflavone and moderate exercise
prevents body fat elevation and bone loss in ovariectomized mice. Metabolism
53, 942–948.
Wu, J., Oka, J., Higuchi, M., Tabata, I., Toda, T., Fujioka, M., Fuku, N., Teramoto,
T., Okuhira, T., Ueno, T., Uchiyama, S., Urata, K., Yamada, K., Ishimi, Y., 2006.
Cooperative effects of isoflavones and exercise on bone and lipid metabolism
in postmenopausal Japanese women: a randomized placebo-controlled trial.
Metabolism 55, 423–433.
Xu, X., Wang, H.J., Murphy, P.A., Cook, L., Hendrich, S., 1994. Daidzein is a more
bioavailable soymilk isoflavone than is genistein in adult women. J. Nutr. 124,
825–832.
Yamashita, T., Sasahara, T., Pomeroy, S.E., Collier, G., Nestel, P.J., 1998. Arterial compli-
ance, blood pressure, plasma leptin, and plasma lipids in women are improved
with weight reduction equally with a meat-based diet and a plant-based diet.
Metabolism 47, 1308–1314.
Yang, G., Shu, X.O., Jin, F., Elasy, T., Li, H.L., Li, Q., Huang, F., Zhang, X.L., Gao, Y.T.,
Zheng, W., 2004. Soyfood consumption and risk of glycosuria: a cross-sectional
study within the Shanghai Women’s Health Study. Eur. J. Clin. Nutr. 58, 615–
620.
Zong, H., Ren, J.M., Young, L.H., Pypaert, M., Mu, J., Birnbaum, M.J., Shulman, G.I.,
2002. AMP kinase is required for mitochondrial biogenesis in skeletal mus-
cle in response to chronic energy deprivation. Proc. Natl. Acad. Sci. U.S.A. 99,
15983–15987.