August 2005: 272283

Special Article

Soybean Foods and Their Benefits: Potential Mechanisms of

Action
Adetayo O. Omoni, MSc, and Rotimi E. Aluko, PhD
Isoflavones have been proposed to be the active component responsible for the beneficial effects of soybean foods, and appear to work in conjunction with
the proteins to protect against cancer, cardiovascular
disease, and osteoporosis. Most of the research activities on the benefits of soybean foods have focused on
the role these isoflavones play in disease prevention or
treatment; however, there is also some evidence that
the benefits are attributable to certain peptides or
protein fractions from soybeans. This review will
focus on some of the potential mechanisms whereby
soybeans exert their protective effects against heart
disease, cancer, and osteoporosis.
Key words: soybean, isoflavones, peptides, cardiovascular disease, cancer
2005 International Life Sciences Institute
doi: 10.1301/nr.2005.aug.272283

INTRODUCTION
Soybean foods have generated a lot of interest recently as a result of evidence that populations consuming
large amounts of soybeans have a lower risk of some
chronic diseases, most notably heart disease and cancer.1-3 Soybean (Glycine max) is an ancient legume that
is traditionally used to prepare both fermented and nonfermented foods, and is a staple among Asian populations. Soybeans are extremely versatile and can be made
into a variety of foods. Asians consume an average of 20
to 80 g of traditional soy foods daily, the most common
of which are tofu, miso, and tempeh.4,5 Americans consume much less soy, only about 1 to 3 g daily,5,6 and this
is mostly in processed forms such as soy drinks, breakfast cereals, energy bars, and soy burgers.
Ms. Omoni and Dr. Aluko are with the Department
of Human Nutritional Sciences, University of Manitoba, Winnipeg, Manitoba, Canada.
Address for correspondence: Dr. Rotimi E. Aluko,
Department of Human Nutritional Sciences, University
of Manitoba, H515 Duff Roblin, Winnipeg, Manitoba,
Canada, R3T 2N2; Phone: 204-474-9555; Fax: 204474-7593; E-mail: alukor@cc.umanitoba.ca.
272

The consumption of soy foods may reduce the risk

of cardiovascular disease and cancer, and this effect is
seen particularly among Asian populations, in whom
consumption of soy foods is high, compared with Western populations, who eat smaller amounts of soy.1-3 In
addition to cancer and heart disease, data suggest that soy
may also reduce the risk of osteoporosis and help to
alleviate menopausal symptoms,3,7,8 both of which are
major health concerns for women. The aim of this review
is to discuss current knowledge relating to the possible
mechanisms by which some of the bioactive components
in soybeans protect against cardiovascular diseases, cancer, and osteoporosis.
BIOACTIVE COMPONENTS OF SOY AND SOY
FOODS
Soy foods contain an array of biologically active
compounds called phytochemicals that may confer important health benefits.9,10 These include: saponins,
phytates, protease inhibitors, phenolic acids, and lecithin,
all known for their anti-cancer potential;4,6 phytosterols,
which are recognized for their cholesterol-lowering effects; isoflavones, which are known for several health
benefits11; and omega-3 fatty acids, which are well recognized for their cardioprotective effects. Of all the
bioactive components of soybeans, isoflavones have attracted the most attention.4,12
Soy Isoflavones
Soybeans, specifically the isolated soy proteins, are
considered a rich source of isoflavones.1,7,13 A 25 g
portion of soy protein contains approximately 50 mg of
isoflavones, although the amounts vary depending on the
variety of bean and the growing conditions.8 Asian
populations consume as much as 80 mg of total isoflavones daily, while in the United States, consumption is
usually not more than 5 mg/d.14,15 Isoflavones exist in
soybean and unfermented soy foods mostly as glycosides
(genistin, diadzin, and, to a lesser extent, glycetin). In
fermented soy foods, isoflavones are in the aglycone
form (genistein, daidzein, and glycetein).16 Genistein is
Nutrition Reviews, Vol. 63, No. 8

the most abundant isoflavone in soybean17 and is proposed to be the most biologically active.6 Soy isoflavones exert both estrogenic and anti-estrogenic effects,
depending on the tissue in which they are acting.12,16,18
They are structurally and functionally similar to 17estradiol, the most potent mammalian estrogen, and are
thus called phytoestrogens.19 They also have non-hormonal effects, including signal transduction and antioxidant activity.19 Isoflavones have been proposed to be the
active component responsible for the beneficial effects of
soy foods.8,20 They appear to work in conjunction with
soy protein to exert anti-carcinogenic, anti-atherogenic,
and anti-osteoporotic effects,7,21 and most of the research
on the benefits of soy foods have focused on the role that
they play in disease prevention or treatment.19,22 There is
also some evidence that these benefits are attributable to
certain peptides or protein fractions from soybeans.
Biotransformation of Isoflavones in the
Intestine
After ingestion, soy isoflavones are biotransformed
in the intestinal tract, a process that is highly dependent
on intestinal bacterial metabolism.9,16,19,23,24 The glycosides diadzin and genistin cannot be absorbed intact into
the peripheral circulation of healthy adults; they have to
be changed to the aglycones genistein and diadzein via
the action of intestinal -glucosidases (Figure 1),16, 23,25
and can be further metabolized into both estrogenic and
anti-estrogenic metabolites.16 The extent to which isoflavone glycosides are bioavailable is therefore dependent
on gut microflora.
POTENTIAL MECHANISMS OF ACTION
Soy Isoflavones and Cardiovascular Disease
Of all the acclaimed benefits of soy foods, perhaps
the most conclusive one is its protective effects against
cardiovascular disease, which is one of the leading
causes of death worldwide,26,27 with elevated levels of
plasma low-density lipoproteins (LDL) and triglycerides

Figure 1. Biotransformation of soy isoflavones in the intestine.

Nutrition Reviews, Vol. 63, No. 8

presenting a higher risk.26 By contrast, high-density

lipoproteins (HDL) are beneficial.28 Postmenopausal
women are at greater risk for cardiovascular disease
because natural or surgical menopause is associated with
elevated levels of circulating total and LDL cholesterol.29 The reduction in blood total and LDL cholesterol
concentrations with the consumption of products containing soy protein has been shown repeatedly in humans
and several animal models (Table 1).30-32 However, the
exact component responsible for this action has yet to be
clearly defined. Isoflavones have been proposed to be the
active ingredient responsible for the hypocholesterolemic effects of soy.14, 33 A previous study suggested
that isoflavone-rich soy protein is considerably more
effective than isoflavone-depleted soy protein, though
this finding is controversial.7
The cholesterol-lowering effect is one of several
proposed mechanisms by which soy reduces the risk of
heart disease.34 In a study to evaluate the effects of
isoflavone-rich and isoflavone-depleted soy protein on
plasma lipid concentrations in postmenopausal, moderately hypercholesterolemic women, isoflavone-rich soy
protein lowered total and LDL cholesterol more than soy
protein depleted of isoflavones, though no significant
differences were observed in HDL cholesterol or triacylglycerol concentrations. This difference in total and
LDL cholesterol lowering between the two soy protein
supplements suggests an effect attributable to the isoflavone-containing fraction.34
Similar results were observed among 18 postmenopausal women with normal and moderately elevated
cholesterol levels in a randomized crossover trial to
assess the hypocholesterolemic effects of soy isoflavones. Subjects were fed isolated soy protein beverage
containing one of three isoflavone levels, 7.1 mg/d
isoflavone (control), 65 mg/d isoflavone (low isoflavone), and 132 mg/d isoflavone (high isoflavone). The
high-isoflavone diet lowered plasma LDL cholesterol by
6.5%, though there were no significant changes in total
or HDL cholesterol, triacylglycerol, apolipoprotein
(Apo) AI, ApoB, lipoprotein(a), or LDL peak particle
diameter. Though small, a decrease of LDL cholesterol
of this magnitude could be associated with a 16% reduction in cardiovascular disease risk.26
A number of explanations have been put forward for
this cholesterol-lowering effect of soy. It has been suggested that soy reduces blood cholesterol levels by reducing cholesterol and bile acid absorption from the
gastrointestinal tract35 and increasing bile acid excretion.36 When this happens, hepatic cholesterol metabolism shifts to provide cholesterol for enhanced bile acid
synthesis, and cholesterol biosynthesis and LDL receptor
activity increase. The result is an increased removal of
cholesterol from the blood via the LDL receptor, thereby
273

274

Nutrition Reviews, Vol. 63, No. 8

Wong et al., 199831

Potter et al., 199830

Mitchell and Collins,

199943

Merz-Demlow et al.,
200027

Wangen et al., 200126

Gardner et al., 200134

Goodman-Gruen and
Kritz-Silverstein,
200132

Human Studies

Reference

Soy Product Fed, Dose, Duration

Outcome

Daily soy isoflavone intake assessed for one Women with moderate and high genistein consumption had
year with questionnaire; average intake of
significantly improved HDL cholesterol
genistein was 1.3 2.4 mg/d, range
013.9 mg/d
Postmenopausal, moderately
Soy protein with trace amounts of
Total and LDL cholesterol concentrations decreased more
hypercholesterolemic
isoflavones (Soy) or 80 mg aglycone
in the Soy group than in the Soy group; no
women (n 94)
isoflavones (Soy) per day for 12 weeks
significant differences in HDL cholesterol and
triacylglycerol concentrations between the groups
Normocholesterolemic and
Daily consumption of soy protein isolate
High-isoflavone diet reduced plasma LDL cholesterol by
mildly hypercholesterolemic
(SPI) beverage powders providing an
6.5% compared with control; low- and high-isoflavone
women (n 18)
average of 7.1 1.1 (control), 65 11
diet reduced LDL:HDL cholesterol by 8.5% and 7.7%,
(low isoflavone), or 132 22 (high
respectively; isoflavone consumption did not
isoflavone) mg isoflavones per day for
significantly affect plasma concentrations of total or
three 93-day periods in a randomized
HDL cholesterol, triacylglycerol, ApoAI, ApoB, or
crossover design
lipoprotein (a) or the LDL peak particle diameter
Normocholesterolemic
SPI consumed as a beverage powder with
High-isoflavone diet lowered LDL cholesterol by 7.6% to
premenopausal women
three levels of soy isoflavones: 10 1.1
10%, total:HDL cholesterol by 10.2%, and LDL:HDL
(n 13)
(control), 64.7 9.4 (low), and 128.7
cholesterol by 13.8%
15.7 (high) mg per day for three
menstrual cycles each in a randomized
crossover design
Healthy men (n 10)
Soy milk (1 L/d) for 4 weeks
No significant effect of soy supplementation on plasma
cholesterol, triglyceride levels, or HDL:LDL cholesterol
ratios compared with controls
Hypercholesterolemic,
Soy protein (40 mg/d) plus moderate and
Non-HDL cholesterol was reduced in both groups fed
postmenopausal women
higher concentrations of isoflavones (1.39
moderate and higher concentrations of isoflavone
(n 66)
and 2.25 mg isoflavone/g protein,
compared with controls; HDL cholesterol increased in
respectively) for 6 months
the two groups; LDL receptor mRNA was reduced in
the two groups compared with controls
Normocholesterolemic
Soy protein incorporated into the National
Plasma concentration of LDL cholesterol and LDL:HDL
(n 13) and
Cholesterol Educational Program Step I diet
cholesterol in both normocholesterolemic and
hypercholesterolemic
(constituting 75% total protein content)
hypercholesterolemic men were significantly lower
(n 13) men
fed for a total of 10 weeks in a randomized
(approximately 6% and 11%, respectively) when soy
crossover design (average daily soy protein
protein diet was consumed
consumption approximately 50 g)

Postmenopausal women
(n 208)

Population (n)

Table 1. Cholesterol-lowering Effects of Soy Products

Nutrition Reviews, Vol. 63, No. 8

275

40

Tsai and Huang, 199910

Lucas et al., 200129

Fukui et al., 200211

Demonty et al., 200233

Adams et al., 200456

Animal Studies

Nestel et al., 1997

Reference

Outcome

Soy isoflavone tablets (80 mg/d) for 5 to 10 No effect on plasma lipids (plasma cholesterol, triglyceride
weeks
and HDL cholesterol); arterial compliance improved by
26%

Soy Product Fed, Dose, Duration

One of five experimental diets: soy plus

Reduced atherosclerosis observed in all mice fed the isoflavones, -conglycinin (7S globulin, a
conglycinin diets compared with mice fed the soy plus
major soy storage protein), isoflavones diet; extent of atherosclerosis reduced in
conglycinin-devoid soy protein, glycinin
ovariectomized female mice fed all soy protein(11S globulin, a major soy storage
containing diets compared with controls
protein), or W008 (a soy peptide fraction)
for 4 months
Male Sprague-Dawley rats
SPI and casein with similar isoflavone
Plasma total triglycerides decreased by 26% in rats fed soy
(n 36)
content (1.82 mg/g protein) for 21 days
protein and casein plus isoflavones compared with
control
Male Sprague-Dawley rats
20% SPI; 20% isoflavone-free SPI (IFSPI) Plasma total cholesterol concentrations of rats fed SPI and
(n 30)
for 2 weeks
IF-SPI were comparable and significantly lower than that
of rats fed control diet; higher fecal steroid excretion in
the SPI group
Ovariectomized female golden Ethanol extracted isoflavone-depleted SPI
ID-SPI lowered serum triglyceride concentrations
Syrian hamsters (n 48)
(IDSPI) (240 g/kg of diet) for 70 days
compared to controls, no significant differences in serum
HDL concentrations, liver total lipids and liver total
cholesterol
Male golden Syrian hamsters SPI containing 1.24 mg genistein and 0.61 SPI group exhibited significantly lower serum total
(n 24)
mg diadzein/g, and ethanol-extracted SPI
cholesterol concentration compared with the ESPI
(ESPI) low in isoflavones (0.21 mg
group; both SPI and ESPI groups had lower LDL
genistein and 0.06 mg diadzein/g) fed for
cholesterol, LDL ApoB, and LDL:HDL cholesterol
10 weeks
compared with the controls

Atherosclerosis-susceptible
mice, male and
ovariectomized female
ApoE-null mice, and LDLreceptor-null ApoB
transgenic mice (n 416)

Menopausal and
perimenopausal women
(n 21)

Population (n)

Table 1. (Contd) Cholesterol-lowering Effects of Soy Products

reducing blood cholesterol levels (particularly the LDL

fraction). There is evidence that soy protein increases
fecal excretion of bile acids. In rats fed isoflavone-rich
and isoflavone-depleted soy protein, higher fecal steroid
excretion was observed in the isoflavone-rich group,
though both groups had lowered total cholesterol compared with the control group. The authors concluded that
the cholesterol-lowering effects of soy may be attributed
to the protein content, with the isoflavones and other
minor constituents playing a role.11
Soy isoflavones are also believed to reduce the risk
of heart disease by reducing the susceptibility of LDL to
oxidation via its antioxidant action.16,37 The effects of a
soy protein diet enriched with isoflavones and one depleted of isoflavones on LDL resistance to oxidation
were compared among 24 healthy subjects in a randomized crossover design. Plasma concentrations of
8-epiprostaglandin F2, a biomarker of lipid oxidation,
were significantly lower after the high-isoflavone diet
(21.2 mg diadzein, 34.8 mg genistein), showing increased resistance of LDL to Cu2-induced oxidation.
This antioxidant action may be significant with regard to
risk of atherosclerosis and cardiovascular disease in
general.16 Similar results were observed in male golden
Syrian hamsters.10 The resistance of LDL to Cu2induced oxidation was greater in hamsters fed isoflavone-rich soy protein than in those fed isoflavonedepleted protein, as assessed by the lower concentrations
of thiobarbituric acid reactive substances, another biomarker of lipid oxidation, and the longer lag time
required for the formation of conjugated dienes. The
isoflavone-enriched group also had significantly higher
serum total antioxidant capacity, particularly -tocopherol content, showing sparing effects on -tocopherol
contents in both serum and liver. These findings suggest
that the intake of soy isoflavones enhanced the resistance
of LDL to oxidation, contributing to antioxidant defense,
and reduced the consumption of -tocopherol in both the
serum and liver of hamsters.10
Improvement of arterial compliance is another possible mechanism by which soy isoflavones protect
against heart disease.38 Arterial compliance (arterial
elasticity) is an important cardiovascular disease risk
factor that diminishes with age and menopause.39 The
effect of soy isoflavones (80 mg daily) on arterial compliance was tested in menopausal and perimenopausal
women over 5- to 10-week periods in a placebo-controlled crossover trial.40 Though there was no effect on
plasma lipids, systemic arterial compliance improved by
26% compared with placebo to about the same extent as
that achieved with conventional hormone replacement
therapy (HRT). The fact that plasma lipids were not
changed suggests that other constituents of soybean
276

(apart from its isoflavones) may be responsible for lipid

lowering.40
Soy Isoflavones and Cancer
The consumption of high levels of soy foods has
been shown to be associated with a reduced risk of
several types of cancer, including breast, endometrial,
and prostate cancers, particularly among Asian populations.5,6,15,41,42 This is in contrast to Western populations, where there is a higher incidence of these cancers.
A number of in vivo studies are supportive of a protective role for soy foods in cancer and some studies have
shown that soy isoflavones are responsible for these
protective effects.15 This protective effect appears to be
strongest for breast cancer. Administration of soy isoflavones early in life enhances the early maturation and
differentiation of the mammary gland of rats, suggesting
that exposure in early life may be important.3,7,17 Table
2 shows a summary of previous studies on the effects of
soy products on cancer and associated DNA damage.
Several possible mechanisms have been identified
for the anticarcinogenic activity of soy isoflavones. One
of the mechanisms by which soy isoflavones are believed
to exert their anticarcinogenic effects is via their antioxidant properties,14 which result in a decrease in lipid
peroxidation16 and oxidative DNA damage,14 both important risk factors for carcinogenesis. These antioxidant
properties are suggested to be related to the chemical
structure of soy isoflavones. Consumption of soy containing naturally occurring amounts of isoflavones reduced lipid peroxidation in vivo among 24 healthy subjects, as evidenced by lower levels of plasma
concentrations of 8-epiprostaglandin F2, a biomarker of
in vivo lipid peroxidation.16 In another study to evaluate
the effects of soy isoflavone supplementation on markers
of oxidative stress in men and women, soy supplementation was given in the form of Novasoy tablets containing 50 mg of isoflavones daily for 3 weeks. There was a
significant decrease in the levels of 5-hydroxymethyl-2deoxyuridine, a biomarker of oxidative DNA damage;
though no significant decrease in lipid peroxidation was
observed. The results from this study suggest that dietary
supplementation with soy isoflavones can decrease levels
of oxidative DNA damage, which is associated with the
process of carcinogenesis.14 A similar decrease in oxidative DNA damage was also observed among men
consuming 1 L of soy milk daily for 4 weeks.43
Another mechanism by which soy isoflavones may
protect against cancer is through the induction of phase II
enzymes,44 which is associated with cancer chemopreventive potential at both the initiation and promotion
phases.44 Induced levels of the phase II enzymes glutathione-s-transferase (GST) and quinone reductase (QR)
Nutrition Reviews, Vol. 63, No. 8

Nutrition Reviews, Vol. 63, No. 8

277

24 healthy subjects
(19 women and
5 men)

Healthy men
(n 10)

Wiseman et al.,
200016

Mitchell & Collins,

199943

Female SpragueDawley rats

(n 119)

Rats (n 150)

Female SpragueDawley rats

(n 86)

Constantinou et al.,
200244

Cohen et al., 20006

Applet and Reicks,

199945

Animal Studies

Men (n 6) and
women (n 6)

Population

Djuric et al., 200114

Human Studies

Reference

One of five experimental diets for 120 days:

genistein (200 mg/kg diet); diadzein (200
mg/kg diet); genistein plus diadzein at
100 mg/kg diet each; 16% soy protein
isolate plus isoflavones (SPI); 16%
isoflavone-depleted SPI (SPI)
20% intact soy protein (SP), 10% SP, 20%
isoflavone-depleted SP (IDSP), 10%
IDSP for 18 weeks
Soy containing three levels of isoflavones
(0.03, 0.4, 0.81 mg/g diet; low, middle
and high level of isoflavones,
respectively) for 13 weeks

One of two experimental diets over two

17-day periods in a randomized crossover
design: low-soy isoflavone diet (21.2 mg
diadzein, 34.8 mg genistein) or high-soy
isoflavone diet (0.9 mg diadzein, 1.0 mg
genistein)
Soy milk (1 L/d) for 4 weeks

Novasoy tablets containing 50 mg

isoflavones; women took 1 tablet daily
and men took 2 tablets daily for 3 weeks

Soy Product Fed, Dose, Duration

Table 2. Effect of Soy Products on Cancer and Oxidative DNA Damage

Rats fed high-isoflavone diet had lower mammary tumor

incidence (40%) compared with controls (71.4%); rats fed
middle and low isoflavone diet had tumor incidence of 53.3
and 57.1%, respectively

No significant differences in mammary tumor incidence, latency,

multiplicity, or volume compared with controls

SPI was the most effective diet, significantly reducing

mammary tumor multiplicity by 50%; diadzein and SPI diet
also significantly reduced tumor multiplicity by 32% compared
with controls; no significant reduction in mammary tumor
incidence in any diet

Decrease in oxidative DNA damage by approximately 67%

Soy isoflavone supplementation decreased levels of oxidative

DNA damage in men and women, evidenced by decreased
levels of 5-hydroxymethyl-2-deoxyuridine, a biomarker of
oxidative DNA damage (47% and 61% decrease in women and
men, respectively)
High-isoflavone diet significantly reduced plasma concentrations
of F2-isoprostane-8-epiprostaglandin F2, a biomarker of in
vivo lipid peroxidation

Outcome

have been proposed as suitable biomarkers for identifying compounds likely to inhibit carcinogenesis.45 QR
and GST are enzymes that help the body get rid of the
toxic products of oxidative metabolism of aromatic hydrocarbons.44 There is evidence to support the mechanism of soy isoflavones as antioxidants and as phase II
enzyme inducers. Soy isoflavones increased antioxidant
and phase II enzyme activity, especially QR, GST, and
UDP-glucuronosyltransferase, in various tissues of rats
fed a high-isoflavone diet for 2 weeks.45
Similar results were found in a study by Constantinou et al.44 comparing the anti-tumor effects of
isoflavone-enriched soy protein isolate and isoflavonedepleted soy protein isolate with those of its isoflavones,
genistein and diadzein. Although the diadzein and isoflavone-enriched soy groups showed significantly reduced
tumor multiplicity compared with the controls, both soy
groups (but not the genistein or diadzein groups) upregulated transcriptional expression of the antioxidant enzymes QR and GST. These results suggest that the mode
of preventive action of soy protein isolate is distinct from
that of the main isoflavones, and that isoflavone-depleted
soy may be more beneficial than isoflavone-enriched soy
in preventing mammary tumors in these experimental
animals.44 However, another study evaluating the effects
of intact and isoflavone-depleted soy protein on Nnitroso-N-methylurea (NMU)-induced rat mammary tumorigenesis showed no effect on tumor incidence, latency, multiplicity, or volume. The results from this
study do not support the hypothesis that the isoflavone
components of soy protein, or the soy protein itself,
inhibit chemically induced mammary tumor development.6
As with animal experiments, studies in human populations relating soy consumption to reduced risk of
breast cancer have produced conflicting results, and a
few have shown stimulatory effects. Petrakis et al.46
reported that prolonged consumption of soy protein isolate by a group of healthy premenopausal US women
increased hyperplastic epithelial cells in duct fluid aspirates, showing a stimulatory effect on the breast.
Soy Isoflavones and Osteoporosis
The ovarian hormone deficiency associated with
menopause results in an increased rate of bone turnover,
which causes an imbalance between bone resorption and
bone formation, thereby accelerating bone loss that leads
to osteoporosis.25,47 HRT is an effective treatment in
reducing the rate of bone loss and risk of fracture, though
not very popular among postmenopausal women because
of the undesirable side effects and long-term risks of
breast and endometrial cancer associated with prolonged
use.18,25,47,48 Soy isoflavones have been shown to alleviate osteoporosis without the side effects of HRT.12 For
278

this reason, soy isoflavones are called selective estrogen

receptor modulators (SERMs), and may be a possible
alternative to HRT.7,12
Soy isoflavones have been suggested to alleviate
osteoporosis by inhibiting bone resorption and stimulating bone formation. Evidence that soy isoflavones reduce
bone resorption has been demonstrated by a number of
studies. Picherit et al.49 investigated the ability of longterm daily intake of soy isoflavones in reversing osteopenia in the adult ovariectomized rat, which was used as a
model of postmenopausal women. Soy isoflavones reduced the urinary excretion of deoxypyridinoline, a specific biomarker of bone resorption.49 Similar results were
also observed among postmenopausal women, in whom
soy protein intake was associated with significantly
lower urinary deoxypyridinoline excretion.50,51
Soy isoflavones appear to stimulate osteoblastic activity through the promotion of insulin-like growth factor-I (IGF-I) production.52 It is well recognized that
IGF-I enhances osteoblastic activity in humans.25 IGF-I
is a protein involved in the bone formation process, and
therefore an increase in IGF-I is indicative of increased
bone formation. In a study by Arjmandi et al.52 using a
rat model of osteopenia, soy increased the gene expression of IGF-I, as indicated by higher femoral mRNA
levels. Incorporation of soy protein with normal isoflavone content (2.3 mg/g protein) had a greater effect on
femoral IGF-I mRNA than the isoflavone-depleted soy
protein-based diet (approximately 0.1 mg/g protein).
This finding indicates that isoflavones may have a role in
enhancing the synthesis of IGF-I at the bone level.
There is additional evidence that soy promotes bone
formation. Increased levels of bone alkaline phosphatase,
a biomarker of bone formation, was observed among
perimenopausal women fed an isoflavone-rich soy protein diet.18 In the same study, lumbar spine bone mineral
density (BMD) and bone mineral content (BMC) were
assessed at baseline and at the end of the treatment.
There was a significant difference in BMD (5.6%) and
BMC (10.1%) between the isoflavone-rich and control
groups, and the authors concluded that soy isoflavones
attenuated bone loss from the lumbar spine in estrogendeficient perimenopausal women.18 Similar results were
also observed among hypercholesterolemic, postmenopausal women assessed for total and regional BMC and
BMD before and after administration of 40 mg/d of soy
protein containing moderate and higher concentrations of
isoflavones (1.39 and 2.25 mg isoflavone/g of protein,
respectively). The high-isoflavone diet significantly increased both BMC and BMD in the lumbar spine but not
elsewhere.30 Increased BMD and mechanical bone
strength and intestinal calcium absorption were also
observed in ovariectomized, osteoporotic rats fed soy
milk.53 Soy is also a rich source of calcium, and this may
Nutrition Reviews, Vol. 63, No. 8

be a possible explanation for the observed increased in

intestinal calcium absorption.
From these studies, it is evident that soy foods play
a role in attenuating bone loss (Table 3); however, the
exact osteoprotective mechanism of soy isoflavones remains unclear and requires further elucidation.
Soy Proteins and Peptides in Cardiovascular
Disease and Cancer
Some evidence exists to support the role of certain
isoflavone-free soy protein and peptide fractions in cardiovascular disease and cancer. Sirtori et al.54 have
shown that 7S globulin, a major storage protein in
soybean, reduced plasma cholesterol concentration by
35% in rats. Similar results were observed by Lovati et
al.55 in an in vitro study evaluating LDL receptor activity
in Hep G2 cells (a human hepatoma cell line) exposed
either to small natural peptides produced by enzymatic
digestion of CroksoyR70 (a commercial isoflavone-poor
soy concentrate) or to synthetic peptides corresponding
to specific sequences of the complete 7S globulin (Table
4). The findings from this study demonstrated that 7S
globulins stimulate the expression of LDL receptors and
the degradation of LDL in the cultured hepatocytes. The
data also support the hypothesis that there are bioactive
peptides produced from the digestion of soy protein that
are absorbed from the small intestine and have beneficial
effects on lipoprotein metabolism and cardiovascular
health by up-regulating LDL-receptor activity in liver
cells. Similar degradation of LDL by an alcohol-extracted 7S globulin was also shown, which demonstrates
that the active molecules are the proteins and not the
alcohol-soluble isoflavones.55 The authors concluded
that the isoflavone-free protein component is likely to be
responsible for this biochemical effect of soy.
To further demonstrate this effect in vivo, Adams et
al.56 assessed the effects of dietary -conglycinin, a major
soy storage protein, and other soy peptide fractions on the
development of atherosclerosis in atherosclerosis-susceptible mice. Male and ovariectomized female ApoE-null mice
and LDL-receptor-null ApoB transgenic mice were randomly assigned to one of six treatment groups differing
only in their source of dietary protein: 1) casein/lactalbumin, 2) isoflavone-containing soy protein isolate, 3) -conglycinin, 4) glycinin (11S globulin, another major soy
storage protein), 5) -conglycinin-devoid soy protein, or 6)
W008 (a peptide fraction produced by hydrolysis and precipitation of soy protein isolate). After 4 months of feeding,
the aortic atherosclerosis (cholesteryl ester content) and
plasma lipoprotein cholesterol concentrations of the mice
were quantified. Results showed that the extent of atherosclerosis was reduced in ovariectomized female mice
fed all soy protein-containing diets compared with
Nutrition Reviews, Vol. 63, No. 8

mice fed casein/lactalbumin-based diets. Furthermore,

compared with mice fed isoflavone-containing soy
protein isolate, atherosclerosis was reduced only in
mice fed the -conglycinin-containing diet. These
reductions were 39% and 67% in male and ovariectomized female ApoE-null mice, respectively, and 66%
in the male LDL-receptor-null mice. These observed
effects were unrelated to variations in isoflavone content of the protein source and only minimally related
to plasma lipoprotein cholesterol concentrations. The
authors thus concluded that a diet rich in -conglycinin has atheroprotective effects that greatly exceed
those of isoflavone-containing soy protein isolate. Interestingly, however, their results show that these
effects do not depend on LDL receptors or influence
plasma lipoproteins.
Evidence also shows that dietary soy peptides and
soy protein isolates (without isoflavones) have antioxidative properties. Takenaka et al.57 demonstrated that
isoflavone-free soy protein isolate and soy peptide reduced paraquat-induced oxidative stress in rats.
Soy protein may also have cancer-protective effects.
Azuma et al.58 and Kanamoto et al.59 demonstrated that
feeding an insoluble, high-molecular weight protein fraction (HMF) prepared from a proteinase-treated soybean
protein isolate suppressed colon and liver tumorigenesis
induced by azoxymethane and dietary deoxycholate in
experimental animals. Bile acid is known to play a
critical role in liver and colon tumorigenesis. The authors
proposed that HMF exerted these protective effects in
colon and liver tumorigenesis by interfering with the
enterohepatic circulation of bile acids, thus inhibiting
resorption of bile acids in the intestine and increasing
fecal bile acid excretion. Peptides found in the feces of
HMF-fed animals were found to be rich in hydrophobic
amino acids, so the authors suggested that the HMF
protein forms non-dissociable complexes with bile acids
in the intestine through hydrophobic binding, which are
then excreted into feces. HMF may therefore be used as
a functional food to prevent the tumor-promoting activity
of bile acids on the liver and colon.
CONCLUSIONS AND FUTURE DIRECTIONS
The consumption of soy foods has been proven to
protect against heart disease, some forms of cancer, and
osteoporosis. However, there is still no conclusive evidence
to date showing whether the protective effects of soy are
derived solely from isoflavones, from soy protein itself, or
from a combination. Research findings are inconsistent;
some evidence supports a role for the isoflavones contained
in soybeans and many soy foods, while other evidence
supports certain peptides or peptide fractions derived from
soy. In addition to this, soy foods also contain other bioac279

280

Nutrition Reviews, Vol. 63, No. 8

Arjmandi et al.,
199852

Ovariectomized rats with

established bone loss
(n 36)

Soy protein with normal isoflavone content

(2.3 mg/g protein) and reduced isoflavone
content (approximately 0.1 mg/g protein) for
65 days

Soybean isoflavone fed as powdered soy

isoflavone concentrate; daily intake of 0, 20,
40 and 80 mg/kg body weight for 84 days

Hypercholesterolemic,
postmenopausal women
(n 66)

Potter et al.,
199830

Adult ovariectomized rats

(n 36)

Soy protein (40 mg/d) plus moderate and

higher concentrations of isoflavones (1.39
and 2.25 mg isoflavone/g protein,
respectively) for 6 months

Perimenopausal women
(n 69)

Alekel et al.,
200018

Animal Studies
Picherit et al.,
200149

Daily soy protein intake assessed over a period

of three consecutive days using a
questionnaire (mean intake 12.6 g/d)
SPI with isoflavones (80.4 mg/d) for 24 weeks

Postmenopausal Japanese women

(n 85)

Human Studies
Horiuchi et al.,
200050

Soy Product Fed, Dose, Duration

Population

Reference

Table 3. Effect of Soy Products on Bone Health

Daily soybean isoflavone consumption decreased

bone turnover as evidenced by decreased urinary
excretion of deoxypyridinoline, a specific bone
marker of bone resorption
Increased expression of femoral insulin-like growth
factor-I mRNA levels, an indicator of increased
bone formation

Soy protein intake significantly correlated with higher

bone mineral density (BMD) and lower urinary
excretion of deoxypyridinoline
Soy isoflavones attenuated bone loss from the lumbar
spine: BMD and bone mineral content (BMC)
increased by 5.6% and 10.1%, respectively,
compared with controls; increased levels of serum
bone-specific alkaline phosphatase, a biomarker of
bone formation
High-isoflavone diet significantly increased both
BMC and BMD in the lumbar spine but not
elsewhere

Outcome

Table 4. Effect of CroksoyR70, its Enzyme Digests and Subfractions of Different Molecular Weight on LDL
Receptor Activity in Hep G2 Cells*
Uptake

Lipoprotein-deficient Serum (LPDS)

CroksoyR70, g/L
0.050
0.125
0.250
0.500
0.750
1.000
CroksoyR70 enzyme digested, g/L
0.050
0.125
0.250
0.500
0.750
1.000
Fractions from CroksoyR70 enzyme digested, g/L
MW 3000 Da
0.050
0.125
0.250
0.500
1000 MW 3000 Da
0.050
0.125
0.250
0.500
MW 1000 Da
0.050
0.125
0.250
0.500

mg
107 12

Degradation
125

I-LDL/g cell protein

117 16

105 9
116 8
132 11
144 7
139 10
108 7

115 10
123 7
145 11
166 8
146 8
120 12

117 11
136 9
153 10
182 13
192 11
220 13

119 11
180 9
191 22
249 19
250 20
266 8

125 11
145 13
169 9
137 10

153 11
190 15
210 12
226 19

89 10
96 12
90 10
88 15

75 11
84 9
88 12
78 19

88 10
95 22
85 10
80 10

77 11
78 9
70 12
75 9

* Values are means SEM, n 3.

P 0.05 compared with LPDS

P 0.0001 compared with LPDS

P 0.05 compared with CroksoyR70 at the same concentration

Reproduced from Lovati et al.55 with copyright permission of the American Society for Nutritional Sciences.

tive components that could exert beneficial effects: the

cholesterol-lowering effects of soluble fiber and phytosterols; the anti-carcinogenic potential of saponins, phytic acid,
Bowman-Birk inhibitor, and lecithin; and the cardioprotective effects of omega-3 fatty acids. Furthermore, these
components may have synergistic effects.
It is evident that there is still a lot to be learned about
soy foods. A question that remains to be answered
conclusively is what component of soy foods is responsible for its protective effects against heart disease,
cancer, and osteoporosis. This gap in knowledge could
form the basis for more research on the benefits of soy,
and further elucidation of its mode of action. Other areas
worthy of research are the possible variations in effects
Nutrition Reviews, Vol. 63, No. 8

of the form of soy foods consumed (fermented, nonfermented) on different tissues and the possible synergistic effects of the many bioactive components of soy.
Furthermore, tissue-specific metabolism and biotransformation of soy isoflavones, and the effects and mechanisms of action of its different metabolites in vivo, will
go a long way in contributing to the understanding of the
mechanisms of action of soy isoflavones.
ACKNOWLEDGEMENT
The authors thank the Natural Sciences and Engineering Research Council of Canada for financial support provided to the research program of Dr. Aluko.
281

REFERENCES
1.

2.

3.

4.

5.

6.

7.

8.

9.

10.

11.

12.

13.

14.

15.

16.

282

Tikkanen MJ, Adlercreutz H. Dietary soy-derived

isoflavone phytoestrogens. Biochem Pharmacol.
2000;60:15.
Kulling SE, Honig DM, Metzler M. Oxidative metabolism of the soy isoflavones daidzein and genistein
in humans in vitro and in vivo. J Agric Food Chem.
2001;49:3024 3033.
Persky VW, Turyk ME, Wang L, et al. Effect of soy
protein on endogenous hormones in postmenopausal women. Am J Clin Nutr. 2002;75:145153.
Messina M, Flickinger B. Hypothesized anticancer
effects of soy: evidence points to isoflavones as the
primary anticarcinogens. Pharmaceutical Biology.
2002;40(suppl):S6 S23.
Fournier DB, Erdman JW Jr, Gordon GB. Soy, its
components, and cancer prevention: a review of the
in vitro, animal, and human data. Cancer Epidemiol
Biomarkers Prev. 1998;7:10551065.
Cohen LA, Zhao Z, Pittman B, Scimeca JA. Effect of
intact and isoflavone-depleted soy protein on NMUinduced rat mammary tumorigenesis. Carcinogenesis. 2000;21:929 935.
Messina M. Soyfoods and soybean phyto-oestrogens (isoflavones) as possible alternatives to hormone replacement therapy (HRT). Eur J Cancer.
2000;36(suppl 4):S71S72.
Genovese MI, Lajolo FM. Isoflavones in soy-based
foods consumed in Brazil: levels, distribution and estimated intake. J Agric Food Chem. 2002;50:5987
5993.
Setchell KDR. Phytoestrogens: the biochemistry,
physiology, and implications for human health of
soy isoflavones. Am J Clin Nutr. 1998;68(suppl 6):
1333S1346S.
Tsai P, Huang P. Effects of isoflavones containing
soy protein isolate compared with fish protein on
serum lipids and susceptibility of low density lipoprotein and liver lipids to in vitro oxidation in
hamsters. J Nutr Biochem. 1999;10:631 637.
Fukui K, Tachibana N, Wanezaki S, et al. Isoflavonefree soy protein prepared by column chromatography reduces plasma cholesterol in rats. J Agric
Food Chem. 2002;50:57175721.
Messina MJ, Loprinzi CL. Soy for breast cancer
survivors: a critical review of the literature. J Nutr.
2001;131(suppl 11):3095S3108S.
Wu AH, Yang D, Pike MC. A meta-analysis of soyfoods and risk of stomach cancer: the problem of
potential confounders. Cancer Epidemiol Biomarkers Prev. 2000;9:10511058.
Djuric Z, Chen G, Doerge DR, Heilbrun LK, Kucuk O.
Effect of soy isoflavone supplementation on oxidative stress in men and women. Cancer Lett. 2001;
172:1 6.
Miltyk W, Craciunescu CN, Fischer L, et al. Lack of
significant genotoxicity of purified soy isoflavones
(genistein, diadzein, and glycitein) in 20 patients
with prostate cancer. Am J Clin Nutr 2003;77:875
882.
Wiseman H, OReilly JD, Adlercreutz H, et al. Isoflavone phytoestrogens consumed in soy decrease
F2-isoprostane concentrations and increase resis-

17.

18.

19.

20.

21.

22.

23.

24.

25.

26.

27.

28.

29.

30.

31.

32.

tance of low-density lipoprotein to oxidation in humans. Am J Clin Nutr. 2000;72:395 400.

Lamartiniere CA, Cotroneo MS, Fritz WA, Wang J,
Mentor-Marcel R, Elgavish A. Genistein chemoprevention: timing and mechanisms of action in murine
mammary and prostate. J Nutr. 2002;132:552S
558S.
Alekel DL, Germain AS, Peterson CT, Hanson KB,
Stewart JW, Toda T. Isoflavone-rich soy protein
isolate attenuates bone loss in the lumbar spine of
perimenopausal women. Am J Clin Nutr. 2000;72:
844 852.
Setchell KD, Brown NM, Zimmer-Nechemias L, et
al. Evidence for lack of absorption of soy isoflavone
glycosides in humans, supporting the crucial role of
intestinal metabolism for bioavailability. Am J Clin
Nutr. 2002;76:447 453.
Schoene NW, Guidry CA. Dietary soy isoflavones
inhibit activation of rat platelets. J Nutr Biochem.
1999;10:421 426.
Anthony MS. Soy and cardiovascular disease: cholesterol lowering and beyond. J Nutr. 2000;130:
662S 663S.
Song TT, Hendrich S, Murphy PA. Estrogenic activity of glycitein, a soy isoflavone. J Agric Food Chem.
1999;47:16071610.
Setchell KD. Soy isoflavones: benefits and risks
from natures selective estrogen receptor modulators (SERMs). J Am Coll Nutr. 2001;20(suppl 5):
354S362S.
Turner NJ, Thomson BM, Shaw IC. Bioactive isoflavones in functional foods: the importance of gut
microflora on bioavailability. Nutr Rev. 2003;61(6
part 1):204 213.
Arjmandi BH, Smith BJ. Soy isoflavones osteoprotective role in postmenopausal women: mechanism
of action. J Nutr Biochem. 2002;13:130 137.
Wangen KE, Duncan AM, Xu X, Kurzer MS. Soy
isoflavones improve plasma lipids in normocholesterolemic and mildly hypercholesterolemic postmenopausal women. Am J Clin Nutr. 2001;73:225
231.
Merz-Demlow BE, Duncan AM, Wangen KE, et al.
Soy Isoflavones improve plasma lipids in normocholesterolemic, premenopausal women. Am J Clin
Nutr. 2000;71:14621469.
Friedman M, Brandon DL. Nutritional and health
benefits of soy proteins. J Agric Food Chem. 2001;
49:1069 1086.
Lucas EA, Khalil DA, Daggy BP, Arjmandi BH. Ethanol-extracted soy protein isolate does not modulate serum cholesterol in golden Syrian hamsters: a
model of postmenopausal hypercholesterolemia. J
Nutr. 2001;131:211214.
Potter SM, Baum JA, Teng H, Stillman RJ, Shay NF,
Erdman JW Jr. Soy protein and isoflavones: their
effects on blood lipids and bone density in postmenopausal women. Am J Clin Nutr. 1998;68(suppl
6):1375S1379S.
Wong WW, Smith EO, Stuff JE, Hachey DL, Heird
WC, Pownell HJ. Cholesterol-lowering effect of soy
protein in normocholesterolemic and hypercholesterolemic men. Am J Clin Nutr. 1998;68(suppl 6):
1385S1389S.
Goodman-Gruen D, Kritz-Silverstein D. Usual diNutrition Reviews, Vol. 63, No. 8

33.

34.

35.

36.

37.

38.
39.

40.

41.

42.

43.

44.

45.

46.

etary isoflavone intake is associated with cardiovascular disease risk factors in postmenopausal
women. J Nutr. 2001;131:12021206.
Demonty I, Lamarche B, Jones PJH. Role of Isoflavones in the hypocholesterolemic effect of soy. Nutr
Rev. 2003;61(6 part 1):189 203.
Gardner CD, Newell KA, Cherin R, Haskell WL. The
effect of soy protein with or without isoflavones
relative to milk protein on plasma lipids in hypercholesterolemic postmenopausal women. Am J Clin
Nutr. 2001;73:738 735.
Demonty I, Lamarche B, Deshaies Y, Jacques H.
Role of soy isoflavones in the hypotriglyceridemic
effect of soy protein in the rat. J Nutr Biochem.
2002;13:671 677.
Lichtenstein AH. Soy protein, isoflavones and cardiovascular disease risk. J Nutr. 1998;128:1589
1592.
Zhang X, Shu XO, Gao Y, et al. Soy food consumption is associated with lower risk of coronary heart
disease in Chinese women. J Nutr. 2003;133:2874
2878.
Clarkson TB. Soy, soy phytoestrogens and cardiovascular disease. J Nutr. 2002;132:566S569S.
Nestel PJ, Pomeroy S, Kay S, et al. Isoflavones from
red clover improve systemic arterial compliance but
not plasma lipids in menopausal women. J Clin
Endocrinol Metab. 1999;84:895 898.
Nestel PJ, Yamashita T, Sasahara T, et al. Soy
isoflavones improve systemic arterial compliance
but not plasma lipids in menopausal and perimenopausal women. Arterioscler Thromb Vasc Biol.
1997;17:33923398.
Le Marchand L. Cancer preventive effects of flavonoids: a review. Biomed Pharmacother. 2002;56:
296 301.
Busby MG, Jeffcoat AR, Bloedon LT, et al. Clinical
characteristics and pharmacokinetics of purified
soy isoflavones: single-dose administration to
healthy men. Am J Clin Nutr. 2002;75:126 136.
Mitchell JH, Collins AR. Effects of soy milk supplement on plasma cholesterol levels and oxidative
DNA damage in men: a pilot study. Eur J Nutr.
1999;38:143148.
Constantinou AI, Lucas LM, Lantvit D, et al. Soy
protein isolate prevents chemically induced rat
mammary tumors. Pharmaceutical Biology. 2002;
40:24 34.
Appelt LC, Reicks MM. Soy induces phase II enzymes but does not inhibit dimethylbenz[a]anthracene-induced carcinogenesis in female rats. J Nutr.
1999;129:1820 1826.
Petrakis NL, Barnes S, King EB, et al. Stimulatory
influence of soy protein isolate on breast fluid secretion in pre- and postmenopausal women. Cancer
Epidemiol Biomarkers Prev. 1996;5:785794.

Nutrition Reviews, Vol. 63, No. 8

47.

48.

49.

50.

51.

52.

53.

54.

55.

56.

57.

58.

59.

Fernandes G, Lawrence R, Sun D. Protective role of

n-3 lipids and soy protein in osteoporosis. Prostaglandins Leukot Essent Fatty Acids. 2003;68:361
372.
Burke GL, Vitolins MZ, Bland D. Soybean isoflavones as an alternative to traditional hormone replacement therapy: are we there yet? J Nutr. 2000;
130:664S 665S.
Picherit C, Bennetau-Pelissero C, Chanteranne B,
et al. Soybean isoflavones dose-dependently reduce bone turnover but do not reverse established
osteopenia in adult ovariectomized rats. J Nutr.
2001;131:723728.
Horiuchi T, Onouchi T, Takahashi M, Ito H, Orimo H.
Effect of soy protein on bone metabolism in postmenopausal Japanese women. Osteoporos Int.
2000;11:721724.
Arjmandi BH, Khalil DA, Lucas EA, et al. Soy protein
with its isoflavones improves bone markers in middle-aged and elderly women. FASEB J. 2000;15:
A728.
Arjmandi BH, Getlinger MJ, Goyal NV, et al. Role of
soy protein with normal or reduced isoflavone content in reversing bone loss induced by ovarian hormone deficiency in rats. Am J Clin Nutr. 1998;
68(suppl 6):1358S1363S.
Omi N, Aoi S, Murata K, Ezawa I. Evaluation of the
effect of soybean milk and soybean milk peptide on
bone metabolism in the rat model with ovariectomized osteoporosis. J Nutr Sci Vitaminol (Tokyo).
1994;40:201211.
Sirtori CR, Even R, Lovati MR. Soybean protein diet
and plasma cholesterol: from therapy to molecular
mechanisms. Ann N Y Acad Sci. 1993;676:188
201.
Lovati MR, Manzoni C, Gianazza E, et al. Soy protein peptides regulate cholesterol homeostasis in
Hep G2 cells. J Nutr. 2000;130:25432549.
Adams MR, Golden DL, Franke AA, Potter SM,
Smith HS, Anthony MS. Dietary Soy -Conglycinin
(7S Globulin) inhibits atherosclerosis in mice. J Nutr.
2004;134:511516.
Takenaka A, Annaka H, Kimura Y, Aoki H, Igarashi
K. Reduction of paraquat-induced oxidative stress
in rats by dietary soy peptide. Biosci Biotechnol
Biochem. 2003;67:278 283.
Azuma N, Machida K, Saeki T, Kanamoto R, Iwami
K. Preventive effect of soybean resistant proteins
against experimental tumorigenesis in rat colon. J
Nutr Sci Vitamonol (Tokyo). 2000;46:2329.
Kanamoto R, Azuma N, Miyamoto T, Saeki T,
Tsuchihashi Y, Iwami K. Soybean resistant proteins
interrupt an enterohepatic circulation of bile acids
and suppress liver tumorigenesis induced by
azoxymethane and dietary deoxycholate in rats.
Biosci Biotechnol Biochem. 2001;65:999 1002.

283