Blood lipid and oxidative stress responses to soy protein with
isoflavones and phytic acid in postmenopausal women1– 4
Heather M Engelman, D Lee Alekel, Laura N Hanson, Anumantha G Kanthasamy, and Manju B Reddy
ABSTRACT
Background: Postmenopausal women are at risk of cardiovascular
disease (CVD) as a result of unfavorable blood lipid profiles and
increased oxidative stress. Soy protein consumption may help pro-
tect against these risk factors.
Objective: Our objective was to ascertain the effect of the soy
protein components isoflavones and phytate on CVD risk in post-
menopausal women.
Design: In a double-blind 6-wk study, 55 postmenopausal women
were randomly assigned to 1 of 4 treatments with soy protein (40 g/d)
isolate (SPI): low phytate/low isoflavone (LP/LI); normal phytate/
low isoflavone (NP/LI); low phytate/normal isoflavone (LP/NI); or
normal phytate/normal isoflavone (NP/NI). Blood lipids (total,
LDL, and HDL cholesterol and triacylglycerol) and oxidative stress
indexes (protein carbonyls, oxidized LDLs, and 8-iso-
prostaglandin-F2␣) were measured at baseline and 6 wk.
Results: The oxidative stress indexes were not significantly affected
by either phytate or isoflavones. Phytate treatment had a minimal but
nonsignificant effect in reducing protein carbonyls and 8-iso-pros-
taglandin-F2␣; the reductions were 6 – 8% and 4 – 6% in the NP/LI
and NP/NI groups and 1– 4% and 3– 4% in the LP/LI and LP/NI
groups, respectively. Similarly, circulating lipids were not signifi-
cantly affected by either phytate or isoflavones. The decline in total
(6%–7% compared with 2%– 4%) and LDL (10%–11% compared
with 3%–7%) cholesterol did not differ significantly between the
normal- and low-isoflavone groups, respectively.
Conclusion: In postmenopausal women, neither phytate nor isofla-
vones in SPI have a significant effect of reducing oxidative damage
or favorably altering blood lipids. Am J Clin Nutr 2005;81:
590 – 6.
KEY WORDS Postmenopausal women, oxidative stress, car-
diovascular disease, soy protein, isoflavones, phytate, blood lipids
INTRODUCTION
Men are usually at higher risk than are women of developing
cardiovascular disease (CVD). After menopause, the risk for
women becomes similar to that for men. The postmenopausal
lack of estrogen may be responsible for affecting antioxidant
defenses, as well as lipid and lipoprotein concentrations (1), both
of which are implicated in the pathogenesis of CVD (2, 3).
Soy protein consumption has been shown to significantly de-
crease serum concentrations of total and LDL cholesterol and
triacylglycerols (4, 5). Many components associated with soy
protein, eg, isoflavones (6), saponins (7), and ␤-conglycinin (7S
590 Am J Clin Nutr 2005;81:590 – 6. Printed in USA. © 2005 American Society for Clinical Nutrition
globulin) protein fractions (8), are reported to have a lipid-
lowering effect. Adding isoflavone-rich soy protein to a low-fat
diet for 3 mo in hyperlipidemic men and postmenopausal women
significantly reduced total and LDL-cholesterol concentrations
(6). Conversely, a 6-mo study with perimenopausal women
found that isoflavone-rich soy protein had no effect on serum
lipid concentrations (9). Conflicting results may be due to subject
Downloaded from ajcn.nutrition.org by guest on March 15, 2016
selection, large interindividual variation, and varied doses
of isoflavones. Hence, the effect of isoflavones on serum lipids in
humans remains unclear. Soy isoflavones may also possess an-
tioxidant properties that protect against LDL oxidation (5, 10)
and improve total antioxidant status (11). The antioxidant effect
of isoflavones may be due to their ability to donate hydrogen
atoms to free radicals, which makes the radicals less reactive
(12). Although it is unclear whether this free radical quenching
happens in vivo, another possible mechanism of isoflavones
may be the enhancement of antioxidant defenses by increasing
antioxidant enzyme concentrations, as was shown in a mouse
model (13).
Phytate, another component of soy, was originally considered
an antinutrient, but it may be beneficial in some conditions.
Phytate may have a stronger ability to quench free radicals than
do isoflavones because of its metal-chelating ability, which ren-
ders the prooxidant metal iron unavailable to participate in the
Fenton reaction and to catalyze hydroxyl radical formation in
vitro (14). Thus, phytate may prevent oxidative damage, such as
lipid peroxidation (15, 16) and may thereby decrease the forma-
tion of atherosclerotic lesions. Although phytate is absorbed in
rats (17), its absorption in humans is very low (18); nevertheless,
it is possible that even small amounts of phytate may protect
against oxidative stress. In addition to its antioxidant activity,
phytate has shown a lipid-lowering effect in rats (19). Reducing
the ratio of zinc to copper (20) may be the mechanism by which
1
From the Department of Food Science and Human Nutrition, Center for
Designing Foods to Improve Nutrition, Iowa State University, Ames, IA
(HME, DLA, LNH, and MBR), and the Department of Biomedical Sciences,
Iowa State University, Ames, IA (AGK).
2
Presented in part at the 5th International Symposium on the Role of Soy
in Preventing and Treating Chronic Disease, September 2003, Orlando, FL.
3
Supported by a USDA Special Grant to the Center for Designing Foods
to Improve Nutrition (Iowa State University, Ames, IA).
4
Reprints not available. Address correspondence to MB Reddy, 1127
HNSB, Department of Food Science and Human Nutrition, Iowa State Uni-
versity, Ames, IA 50011. E-mail: mbreddy@iastate.edu.
Received March 3, 2004.
Accepted for publication November 16, 2004.
 CVD RISK FACTOR RESPONSE TO SOY PROTEIN
phytate lowers lipid concentrations (21); however, human data in
this area are limited. The overall hypothesis of the current study
was that soy protein would reduce the risk of CVD—specifically,
that soy isoflavones would favorably alter blood lipid concen-
trations and that phytate would decrease oxidative stress indexes.
SUBJECTS AND METHODS
Study design
In this 6-wk double-blind study, 55 free-living postmeno-
pausal women were randomly assigned to 1 of 4 soy protein
isolate (SPI) treatments provided by The Solae Company (St
Louis, MO): low-phytate/low-isoflavone (LP/LI; n ҃ 14),
normal-phytate/low-isoflavone (NP/LI; n ҃ 13), low-phytate/
normal-isoflavone (LP/NI; n ҃ 14), or normal-phytate/normal-
isoflavone (NP/NI; n ҃ 14) treatment. The 2 treatments contain-
ing isoflavone concentrations that are referred to as “normal” are
in fact rich in isoflavones because their isoflavone content per
gram protein is twice that found in typical SPIs. The Solae Com-
pany prepared the low-isoflavone and low-phytate SPIs by using
alcohol extraction and phytase hydrolysis, respectively. The
phytate and isoflavone (aglycone) contents, respectively, of each
treatment per 40 g soy protein were as follows: LP/LI treatment,
0.22 g and 1.2 mg; NP/LI treatment, 0.64 g and 1.2 mg; LP/NI
treatment, 0.22 g and 85.8 mg; and NP/NI treatment, 0.78 g and
84.6 mg. All subjects were white except one Asian woman. The
women were supplied with protein in a powder form in two 20-g
packets/d (84 total packets), and they were given recipes for
incorporating the powder into fruit smoothies or other foodstuffs.
Beginning 2 wk before and continuing throughout the interven-
tion, subjects were required to avoid all supplements, including
vitamins, minerals, and herbal remedies. Subjects were also pro-
vided with a list of foods that are phytate rich (ie, cereals, le-
gumes, and nuts; 22, 23) or isoflavone rich (ie, primarily le-
gumes; 24) and were instructed to avoid these foods during the
intervention.
The study protocol, consent form, and subject-related materi-
als were approved by Human Subjects Review Committee of
Iowa State University (Institutional Review Board ID #02–351).
Written informed consent was obtained from all subjects.
Subject selection
Postmenopausal women were recruited throughout central
Iowa from April through November 2002 by means of campus
and local newspaper advertisements and a television feature
story. Approximately 300 women responded, and the potential
participants were screened by telephone interviews to ensure that
they met the inclusion and exclusion criteria. Health status with
respect to chronic diseases (ie, arthritis, cancer, CVD, diabetes,
or gastrointestinal, kidney, liver, parathyroid, and pulmonary
disease) and menopausal state was assessed before a woman’s
inclusion in the study with the use of a health and medical history
questionnaire. Women were included in the study if they were
postmenopausal (last menses 욷12 mo before intervention) and
healthy (no chronic disease or medication use) and if they had a
body mass index (BMI; in kg/m2) of 19 –34. Women were ex-
cluded if they had a chronic disease, had undergone a hysterec-
tomy, had taken hormone therapy 울12 mo before the interven-
tion, or had used cigarettes or hormone creams 울6 mo before the
591
intervention. On the basis of these criteria, 57 women were qual-
ified to participate. Two of these women withdrew because of
intolerable gastrointestinal side effects. The remaining 55
women completed the 6-wk intervention with minimal or no
gastrointestinal side effects.
Data collection
Health and medical history, nutrition history, and soy food
intake data (25) were obtained at baseline by using interviewer-
administered questionnaires. Usual dietary intake was also as-
sessed at baseline with the use of a food-frequency questionnaire
from Block Dietary Data Systems (Berkeley, CA). Overnight-
fasted blood samples were collected at baseline and week 6 and
were frozen at Ҁ80 °C until they were used. A standard reference
laboratory (Quest Diagnostics, St Louis, MO) analyzed serum
and plasma for the blood lipid profile (total, LDL-, and HDL-
cholesterol and triacylglycerol concentrations) and other blood
chemistry analytes. Indexes of oxidative stress [ie, protein car-
bonyls, oxidized LDL (oxLDL), and 8-iso-prostaglandin-F2␣
(PGF2␣)] were measured in our laboratory at Iowa State Univer-
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sity. Protein carbonyls were measured by using a slight modifi-
cation of the procedure described in Reznick et al (26). Briefly,
plasma was mixed with dinitrophenylhydrazine dissolved in hy-
drochloric acid, accompanied by blanks in hydrochloric acid
alone. Protein was then precipitated with 20% (wt:vol) trichlo-
roacetic acid and washed once with 10% trichloroacetic acid and
3 times with 5 mL of a 1:1 mixture of ethanol and ethyl acetate.
Finally, precipitates were dissolved in a solution of 6 mmol
guanidine-hydrochloric acid/L. The absorbance was measured
spectrophotometrically at 380 nm. Plasma oxLDL concentra-
tions were determined by using an enzyme-linked immunosor-
bent assay kit from ALPCO Diagnostics (Windham, NH), and
serum PGF2␣ (free ѿ esterified) concentrations were determined
by using an enzyme-linked immunosorbent assay kit from
Stressgen Biotechnologies (Victoria, Canada). The overall CVs
for protein carbonyls, PGF2␣, and oxLDL were 쏝1%, 8%, and
6%, respectively.
Statistical analysis
Statistical analysis was performed with SAS software (version
8.0; SAS Institute, Cary, NC) with significance set at P 울 0.05.
The sample size per group was based on a pooled SD of 22 mg/dL
and 80% power (P 쏝 0.05) to detect a reduction in LDL choles-
terol of 15 mg/dL, which is biologically significant enough to
reduce CVD risk. Analysis of variance with Tukey’s multiple
comparison was used to test the differences in baseline values
among the treatments. To ascertain whether baseline-adjusted
differences (6 wk Ҁ baseline) between isoflavone and phytate
treatments differed significantly, we used two-way analysis of
covariance with the respective baseline values as covariates.
Pearson’s product-moment correlation analysis was used to de-
termine the relation among the risk factors at baseline.
RESULTS
Compliance
Compliance was based on the number of packets returned at
the week 6 visit. Most of the women consumed 100% of their
packets. However, 4 subjects returned 2, 3, 4, and 8 packets,
respectively, and 2 subjects requested 3 and 4 extra packets,
592 ENGELMAN ET AL

TABLE 1
Baseline characteristics of subjects1

Treatment group

LP/LI NP/LI LP/NI NP/NI

(n ҃ 14) (n ҃ 13) (n ҃ 14) (n ҃ 14)

Age (y) 56 (49–70) 59 (53–69) 58 (47–72) 60 (50–70)

Weight (kg) 71 (58.2–93.6) 72.7 (59.0–96.8) 72.7 (55.2–92.5) 69.2 (52.3–92.4)
Height (m) 1.7 (1.5–1.8) 1.6 (1.5–1.7) 1.7 (1.6–1.7) 1.7 (1.6–1.7)
BMI (kg/m2) 25.9 (18.6–32.9) 27.9 (21.2–33.9) 26.5 (19.9–32.0) 25.3 (21.3–33.6)
Dietary intake/d2
Energy (kcal) 1702 (515–2559) 1766 (763–2708) 1769 (877–2827) 1647 (1144–2595)
Protein (g) 65 (22–104) 68 (30–108) 71 (31–122) 63 (39–103)
Carbohydrate (g) 217 (53–412) 239 (119–405) 213 (85–339) 204 (146–328)
Fat (g) 66 (25–96) 64 (22–113) 75 (36–139) 67 (30–105)
Saturated fat (g) 21 (7–37) 18 (6–30) 21 (12–38) 18 (9–26)
Vitamin C (mg) 103 (42–192) 155 (99–312) 130 (43–211) 116 (50–170)
Vitamin E (␣-TE) 9 (3–15) 11 (5–19) 11 (6–19) 10 (5–21)
Vitamin A (RE) 1753 (305–3934) 1734 (565–3065) 1570 (698–5107) 1161 (656–2238)
1
All values are x៮ ; range in parentheses. LP/LI, low phytate/low isoflavone; NP/LI, normal phytate/low isoflavone; LP/NI, low phytate/normal isoflavone; NP/NI,
normal phytate/normal isoflavone; ␣-TE, ␣-tocopherol equivalents; RE, retinol equivalents. There were no significant differences between the treatment groups

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(ANOVA).
2
Selected nutrients were assessed by using a food-frequency questionnaire.

respectively. These subjects were evenly distributed across the
treatment groups. Compliance was also checked by randomly
analyzing isoflavone excretion in urine before and after inter-
vention (n ҃ 4 per treatment). The subjects in the LP/NI and
NP/NI treatment groups excreted 23 and 29 ␮mol isoflavone/L,
and the subjects in the LP/LI and NP/LI treatment groups ex-
creted 1.6 and 2.8 ␮mol isoflavone/L, respectively. The graduate
research assistant was in contact with these women on a regular
basis. Fifteen of the 55 subjects reported intermittent constipa-
tion; we provided guidelines to increase fluid intake, which cor-
rected the problem and apparently did not affect compliance.
Subject characteristics
The subjects ranged in age from 47 to 72 y. The time since
menopause ranged from 1 to 20 y (x៮ : 6.4 y). Thirteen subjects had
smoked in earlier years, 6 subjects had experienced cancer (skin,
n ҃ 4; breast, n ҃ 1; and cervical, n ҃ 1), but all 6 were in
remission, and 1 subject had 2 previous minor strokes that were
not related to atherosclerotic CVD. There were no other reported
cardiovascular events and no diagnoses of CVD.
Twenty-eight and 26 of the subjects reported their health to be
good and excellent, respectively, whereas 1 subject reported her
health to be fair. Education levels were high school (n ҃ 16),
college (n ҃ 29), and graduate school (n ҃ 10). Thirteen of the
participants stated they had experienced iron deficiency at least
once in the past as a result of malnutrition, pregnancy, menstru-
ation, or the onset of menopause; however, only one woman had
a baseline hemoglobin concentration 쏝 12 g/dL. Thirty-six sub-
jects (8 –10/treatment) reported regular use of a multivitamin,
and 2 subjects (LP/LI, n ҃ 1; LP/NI, n ҃ 1) reported regular use
of iron supplements. Thirty-five subjects (7–11/treatment) re-
ported they were consuming soy or soy products before the in-
tervention, although most indicated that this consumption was
irregular.
The subjects’ descriptive characteristics and daily nutrient
intakes are reported in Table 1. None of the descriptive charac-
teristics at baseline differed significantly between the treatment
groups. Neither body weight nor BMI differed significantly
among the treatment groups even at 6 wk (data not shown). The
dietary intakes of macronutrients were within normal ranges and
did not differ significantly among the treatment groups. Overall,
intakes of selected antioxidants (vitamins A, C, and E) were
within the recommended allowances in each treatment group.
Although vitamin C intakes were higher in 2 treatments (NP/LI
and LP/NI), the differences were not significant.
Oxidative stress indexes
Oxidative stress indexes before and after intervention and the
mean changes for each treatment are shown in Table 2. The mean
protein carbonyl concentration at baseline was very low (0.2
nmol/mg protein), and there were no significant differences
among the treatment groups. Phytate treatment had a very modest
(P ҃ 0.15) and nonsignificant effect on protein carbonyl con-
centrations, which decreased by 6%– 8% and 1%– 4%, respec-
tively, in the normal-phytate and low-phytate groups. At base-
line, the PGF2␣ concentration in the NP/LI group was
significantly (P ҃ 0.05) different from that in the LP/LI group but
not from that in the other 2 groups. Neither the phytate nor the
isoflavone treatment had a significant effect on PGF2␣ concen-
trations. The reduction in PGF2␣ in the NP/LI group was 6%, and
that in the other 3 groups was 3%– 4%. The mean oxLDL con-
centration at baseline ranged from 66 to 81 U/L across the treat-
ments. As was seen with PGF2␣, there was a 5%–13% decline in
oxLDL across the groups, but this reduction was not significantly
affected by either phytate or isoflavone treatment. The reduction
in oxLDL in the normal-isoflavone groups was 6 –10 U/L, and
that in the low-isoflavone groups was 4 U/L. No significant
phytate ҂ isoflavone interaction was found for any of the above
3 oxidative stress indicators.
Blood lipid concentrations
Blood lipid values at baseline and 6 wk and the mean changes
are shown in Table 2. Mean total cholesterol did not differ across
 CVD RISK FACTOR RESPONSE TO SOY PROTEIN 593
TABLE 2
Oxidative stress and blood lipid measures at baseline and 6 wk1

P2

LP/LI NP/LI LP/NI NP/NI Phytate ҂

(n ҃ 14) (n ҃ 13) (n ҃ 14) (n ҃ 14) Phytate Isoflavone isoflavone

PC (nmol/mg)
Baseline 0.204 앐 0.543 0.197 앐 0.55 0.183 앐 0.39 0.190 앐 0.35
6 wk 0.196 앐 0.49 0.181 앐 0.40 0.185 앐 0.42 0.178 앐 0.43
Change4 Ҁ0.009 앐 0.031 Ҁ0.016 앐 0.030 0.002 앐 0.031 Ҁ0.012 앐 .020 0.15 0.64 0.84
PGF2␣ (pg/mL)
Baseline 4118 앐 573a 5327 앐 1782b 4471 앐 1183a,b 4678 앐 1366a,b
6 wk 3978 앐 654 5000 앐 1200 4286 앐 1040 4411 앐 992
Change Ҁ139 앐 653 Ҁ327 앐 1396 Ҁ186 앐 893 Ҁ207 앐 637 0.29 0.81 0.40
oxLDL (U/L)
Baseline 66.4 앐 15.9 81.0 앐 25.3 77.9 앐 29.4 75.1 앐 21.4
6 wk 62.2 앐 17.9 76.6 앐 24.4 67.9 앐 18.8 69.3 앐 24.0
Change Ҁ4.2 앐 9.2 Ҁ4.3 앐 17.6 Ҁ10.0 앐 17.5 Ҁ5.9 앐 18.5 0.33 0.48 0.87
Cholesterol
Total (mg/dL)
Baseline 223 앐 31 225 앐 31 227 앐 35 235 앐 40
215 앐 33 221 앐 36 213 앐 30 220 앐 38

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6 wk
Change Ҁ8 앐 18 Ҁ4 앐 31 Ҁ14 앐 23 Ҁ16 앐 20 0.67 0.24 0.68
LDL (mg/dL)
Baseline 138 앐 27 139 앐 32 144 앐 30 151 앐 38
6 wk 129 앐 29 135 앐 28 131 앐 27 134 앐 36
Change Ҁ9 앐 19 Ҁ4 앐 33 Ҁ14 앐 16 Ҁ16 앐 16 0.70 0.28 0.61
HDL (mg/dL)
Baseline 63 앐 17a,b 52.6 앐 10b 62 앐 13a,b 65 앐 11a
6 wk 65 앐 14 53.3 앐 10 59 앐 14 64 앐 14
Change 3앐7 0.7 앐 6 Ҁ3 앐 9 Ҁ1 앐 7 0.73 0.23 0.12
Triacylglycerol (mg/dL)
Baseline 112 앐 69a 168 앐 95b 104 앐 43a 100 앐 51a
6 wk 101 앐 55 167 앐 77 117 앐 63 106 앐 77
Change Ҁ11 앐 47 Ҁ0.8 앐 56 Ҁ12 앐 32 7 앐 39 0.52 0.51 0.24
1
LP/LI, low phytate/low isoflavone; NP/LI, normal phytate/low isoflavone; LP/NI, low phytate/normal isoflavone; NP/NI, normal phytate/normal
isoflavone; PC, protein carbonyl; PGF2␣, 8-iso-prostaglandin-F2␣; oxLDL, oxidized LDL. Mean baseline values with different superscript letters are signif-
icantly different (ANOVA).
2
Baseline-adjusted differences (6 wk Ҁ baseline) were compared by using two-way analysis of covariance with respective baseline values as covariates.
3
x៮ 앐 SD (all such values).
4
Changes reflect 6 wk Ҁ baseline values.

treatments at baseline, ranging from 223 to 235 mg/dL. Although
both the normal- and low-isoflavone treatment groups experi-
enced a modest decline in total cholesterol (6%–7% and 2%– 4%,
respectively) after 6 wk, the reductions did not differ signifi-
cantly between the groups. Similarly, LDL cholesterol decreased
with treatment in all groups, but the differences between low and
normal treatments (phytate or isoflavones) were too small to
detect significance. The reduction in LDL cholesterol was 10%–
11% (14 –16 mg/dL) in the normal-isoflavone and 3%–7% (4 –9
mg/dL) in the low-isoflavone group. The mean baseline triacyl-
glycerol concentration in the NP/LI group was significantly
higher than that in the other treatment groups, but we found no
significant effect of treatment. Similarly, HDL cholesterol did
not respond to treatment. As we have noted with oxidative stress,
phyate and isoflavones had no significant interactive effect on
blood lipids.
Correlation of CVD risk factors
Correlations among baseline BMI, oxidative stress indexes,
and blood lipid measures are shown in Table 3. The highest
positive correlations (P 쏝 0.0001) were observed between tri-
acylglycerol and PGF2␣, as well as between LDL cholesterol and
oxLDL and between total cholesterol and oxLDL. In addition,
BMI was highly correlated with triacylclygerol (P 쏝 0.001),
PGF2␣ (P 쏝 0.05), and oxLDL (P 쏝 0.05). HDL cholesterol was
negatively correlated with triacylglycerol (P 쏝 0.0001), BMI
(P 쏝 0.0001), PGF2␣ (P 쏝 0.001), and oxLDL (P 쏝 0.05).
DISCUSSION
The Food and Drug Administration approved the health claim
that a daily intake of 25 g soy protein (27) reduces heart disease
risk. In the current study, we chose to use 40 g soy protein/d to
ascertain whether a higher intake would elicit beneficial effects
on blood lipid profiles, as well as on oxidative stress indexes, in
postmenopausal women, who are at high risk for CVD. This
amount was also chosen on the basis of previous human studies
that investigated health benefits of soy protein (9, 28 –30). More-
over, because of phytate’s low absorption in humans, the subjects
needed to consume 40 g soy protein/d to obtain enough dietary
594 ENGELMAN ET AL
TABLE 3
Intercorrelations of baseline values1
PGF2␣ oxLDL Total
PC 0.15 0.12 0.03
PGF2␣ — 0.25 0.17
oxLDL — — 0.613
Cholesterol
Total — — — 0.943
LDL — — — —
HDL — — — —
Triacylglycerol — — — —
1
Pearson’s correlation coefficient. PC, protein carbonyl; PGF2␣, 8-iso-prostaglandin-F2␣; oxLDL, oxidized LDL.
2
P 쏝 0.001.
3
P 쏝 0.0001.
4
P 쏝 0.05.
5
P 쏝 0.01.
phytate to show beneficial effects. However, the amount of
phytate provided daily from 40 g soy protein (ie, 0.64 – 0.78 g)
was considerably lower than the amount used in an earlier human
study, in which 8.8 g phytate/d was given to patients at risk of
kidney stones (31).
Our results showing no significant effect of soy isoflavones on
oxLDL do not agree with the results of a study by others that
showed a positive correlation between reductions in oxLDL and
serum daidzein, genistein, and total isoflavone concentrations
after a 12-wk intervention with soy foods in postmenopausal
women (5). Perhaps differences in response to treatment may
depend on the menopausal status of the person at baseline and
whether baseline values are taken into account in the analyses, as
we have done. Women in our study were considered to have
normal antioxidant status because the oxLDL concentration in
only 1 woman was greater (170 U/L) than the reference range
(26 –117 U/L) specified by the assay kit manufacturer. However,
our data indicate a strong correlation between oxLDL and blood
lipids, which suggests that a response to treatment may also
depend on the interaction between blood lipids and oxidative
stress. Jenkins et al (10) showed that the consumption of 33 g soy
protein/d for 1 mo significantly decreased oxLDL concentrations
in hyperlipidemic subjects. Although 14 of 55 women in our
study had LDL-cholesterol concentrations high enough that they
could be classified as hyperlipidemic (LDL cholesterol 쏜160
mg/dL; 32), the mean LDL-cholesterol concentrations were
within the normal range (138 –151 mg/dL). Thus, it is not sur-
prising that no significant effect of isoflavone treatment on ox-
LDL concentrations was seen in these relatively normolipidemic
women.
Dent et al (9) had results similar to ours, reporting no effect on
circulating lipids in either normal or hyperlipidemic perimeno-
pausal women fed isoflavone-rich soy protein for 24 wk. This
lack of effect was attributed to the perimenopausal status of these
subjects, whose hormonal milieu is typically erratic. Further-
more, Hsu et al (33) found that supplementing normal postmeno-
pausal women with 150 mg isoflavones/d for 6 mo resulted in no
significant change in plasma lipids. These data, similar to those
for oxidative stress, suggest that the effect of isoflavone depends
on the lipidemic state of each person. It is been reported that the
Cholesterol
LDL HDL Triacylglycerol BMI
Ҁ0.05 Ҁ0.01 0.18 0.10
0.04 Ҁ0.442 0.743 0.334
0.623 Ҁ0.324 0.395 0.344
0.07 0.20 0.14
Ҁ0.06 0.05 0.17
— Ҁ0.653 Ҁ0.523
— — 0.452
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consumption of 60 g soy protein/d for 12 wk had a more pro-
nounced effect on blood lipids in hypercholesterolemic than in
normolipidemic postmenopausal women (4). In addition, the
consumption of isoflavone-rich soy food diets for 1 mo has been
shown to reduce blood lipids and homocysteine concentrations in
hyperlipidemic men and postmenopausal women (6).
Isoflavones in SPI had no effect on HDL cholesterol, which
contradicted a previous finding of a positive association between
soy isoflavone intake and HDL-cholesterol concentrations in
postmenopausal women (34). At baseline, the mean triacylglyc-
erol concentration in the NP/LI group (ie, 100 –112 mg/dL) was
significantly higher than that in the other groups, and yet we
found no effect of treatment in these women with normal tria-
cylglycerol concentrations. Similarly, no change in triacylglyc-
erol concentration was shown in a 24-wk study by feeding a diet
with 30 or 50 g soy protein/d to men and women (35). Hence, the
CVD risk protection of SPI in normolipidemic persons may not
be due to the effect of isoflavones on increasing HDL cholesterol
or reducing triacylglycerol.
Phytate is considered as an antioxidant because it has been
shown in vitro to have a dose-dependent quenching effect on
iron-induced free radical formation (14). The modest (P 쏝 0.15)
decline we found in protein carbonyl concentrations and the
nonsignificant decline in lipid oxidation indicators with phytate
treatment do not support the antioxidant potential of phytate, and
no data are available to suggest the degree of reduction in oxi-
dative stress that is biologically significant. Furthermore, our
results indicating the lack of lipid-lowering effect of phytate do
not support the findings in rat studies (19, 20). However, Jari-
walla et al (19) used in rats a cholesterol-enriched diet with a
considerably higher amount of phytate than our subjects con-
sumed. Nonetheless, the lack of effect we report may be due
either to insufficient amounts of phytate in the treatment or to
phytate’s poor absorption in humans (18) compared with that in
rats (17).
Oxidative stress indicators and the circulating lipids (except
HDL cholesterol) declined in all 4 groups (albeit not signifi-
cantly), but this decline in turn made the differences between the
phytate and isoflavone groups very small and, indeed, too small
 CVD RISK FACTOR RESPONSE TO SOY PROTEIN
to detect significance. For example, we based our study on de-
tecting a 15 mg/dL difference in LDL, but the actual difference
between the normal- and low-isoflavone groups was only 9.5
mg/dL (baseline-adjusted difference of 6.5 mg/dL). It is possible
that other components in SPI might have an additional effect on
the outcome variables. Beneficial effects on lipids and antioxi-
dant status have been shown by other components of soy protein,
such as saponins (7), 7S globulin (8), and arginine (36). Thus, a
single component in SPI may not necessarily be responsible for
protection against CVD, but rather multiple components of SPI
may be protective through different mechanisms.
Although subjects were recruited throughout the summer and
fall seasons, we could not document differences among the treat-
ments at baseline or intervention-related changes in BMI, which
suggests that seasonal variations in dietary intake had no effect
on BMI. Moreover, because all subjects consumed the same
amount of protein during the intervention, the differences in
dietary intakes between the treatment groups were not expected.
However, the positive correlation of BMI with the oxidative
stress markers oxLDL and PGF2␣ (Table 3) is noteworthy. Va-
sankari et al (37) found a similar correlation between BMI and
oxLDL, whereas Suzuki et al (38) found no correlation between
these 2 markers. Baseline BMI was also negatively correlated
with HDL cholesterol, much as was reported in another study
(39). A negative correlation between HDL cholesterol and fat
mass (40) suggests unfavorable HDL-cholesterol concentrations
in overweight or obese persons. Thus, reducing BMI may help to
reduce oxidative stress and maintain HDL cholesterol. However,
confirmation of these results will require further study of the
relation between fat mass and oxidative stress.
In contrast to the studies that examined only total antioxidant
status (11, 30), our study included 3 oxidative stress indicators to
assess specific damage to proteins and lipids, which may be a
better indication of oxidative damage. Yet, we have not shown a
significant effect on these 3 indexes by SPI that contained either
phytate or isoflavones. Previous studies (6, 11) had led us to
believe that a 6-wk intervention was sufficiently long to note
changes in oxidative stress and lipidss. We found no significant
beneficial effect of phytate or isoflavones in this study, but future
studies with higher doses of these components and a greater
number of subjects, perhaps including subjects with hyperlipid-
emia, may provide different results.
We thank Philip Dixon, Department of Statistics at Iowa State University,
for statistical advice.
DLA and MBR participated in the study design. HME and LNH collected
and analyzed the data. HME wrote the first draft of the manuscript, and DLA,
AGK, and MBR provided advice and consultation on the final draft. None of
the authors had any personal or financial conflicts of interest.
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