Journal of the American College of Nutrition

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Whey Protein Supplementation Improves Body

Composition and Cardiovascular Risk Factors in
Overweight and Obese Patients: A Systematic
Review and Meta-Analysis

Kamonkiat Wirunsawanya, Sikarin Upala, Veeravich Jaruvongvanich &

Anawin Sanguankeo

To cite this article: Kamonkiat Wirunsawanya, Sikarin Upala, Veeravich Jaruvongvanich &
Anawin Sanguankeo (2017): Whey Protein Supplementation Improves Body Composition and
Cardiovascular Risk Factors in Overweight and Obese Patients: A Systematic Review and Meta-
Analysis, Journal of the American College of Nutrition, DOI: 10.1080/07315724.2017.1344591

To link to this article: http://dx.doi.org/10.1080/07315724.2017.1344591

Published online: 31 Oct 2017.

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 JOURNAL OF THE AMERICAN COLLEGE OF NUTRITION
https://doi.org/10.1080/07315724.2017.1344591

Whey Protein Supplementation Improves Body Composition and Cardiovascular Risk

Factors in Overweight and Obese Patients: A Systematic Review and Meta-Analysis
Kamonkiat Wirunsawanyaa, Sikarin Upalab,c, Veeravich Jaruvongvanicha, and Anawin Sanguankeo c,d

a
Department of Internal Medicine, University of Hawaii, Honolulu, Hawaii, USA; bSection of Endocrinology, Diabetes, and Metabolism, Department of
Medicine, University of Chicago, Chicago, Illinois, USA; cDepartment of Preventive and Social Medicine, Faculty of Medicine Siriraj Hospital, Mahidol
University, Bangkok, Thailand; dDivision of Nephrology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

ABSTRACT ARTICLE HISTORY

Background: Previous literature shows possible benefits of whey protein supplementation in promoting Received 31 March 2017
weight loss. However, most studies do not have enough power to show beneficial effects on body Accepted 16 June 2017
Downloaded by [Southern Cross University] at 08:03 02 November 2017

composition and cardiovascular disease (CVD) risk factors. This meta-analysis evaluated effects of whey KEYWORDS
protein in individuals who are overweight and obese. Body composition;
Methods: We comprehensively searched the databases of MEDLINE, Embase, and Cochrane databases. The cardiovascular disease; meta-
inclusion criteria were published randomized control trials (RCTs) comparing whey protein analysis; obesity; whey
supplementation to placebo or controls in individuals who are overweight or obese. The primary outcome protein
was the differences in the change in body composition (body weight, waist circumference, total fat mass,
body lean mass). We also examined the changes in CVD risk factors as secondary outcomes. We calculated
pooled mean difference (MD) with 95% confidence intervals (CIs) using a random effects model.
Results: Nine RCTs were included in the meta-analysis. There was a significant reduction of body weight
(MD D 0.56, 95% CI: 0.30–0.81), lean mass (MD D 0.77, 95% CI: 0.59–0.96), and fat mass (MD D 1.12, 95%
CI: 0.77–1.47) favoring the whey protein group. There were improvements in multiple CVD risk factors
including levels of systolic blood pressure, diastolic blood pressure, glucose, high-density lipoprotein, and
total cholesterol (all p values <0.05).
Conclusions: Whey protein supplementation seems to improve body weight, total fat mass, and some CVD
risk factors in overweight and obese patients. Further studies regarding optimal dosage and duration of
whey protein supplementation would be helpful to assess potential favorable effects in individuals who
are overweight or obese.

Introduction
In the past decade, whey protein supplementation has been
widely investigated as a potential intervention to improve car-
diovascular risk factors and body composition, but most of the
previous studies do not have enough power to determine the
biological advantages of whey protein on cardiovascular risk
factors and body composition. While lowering cholesterol or
weight loss medications have adverse side effects, whey protein
supplementation is a pure source of protein that has been shown
to be safe and possibly have potential functions of reducing car-
diovascular risk factors and improving body composition (1–3).
We selectively collected nine studies for a meta-analysis to
analyze the correlation of whey protein supplementation and
improvement of cardiovascular risk factors and body composi-
tion in individuals who are obese and overweight.
Whey protein is a biologically active protein that has been
found to have many potential biological advantages including
cardiovascular benefits, antioxidant properties, enhanced imm-
une function, weight reduction, and maintenance of catabolism
of muscle mass during exercise because it contains many
functional and nutritional components, especially essential and
branched-chain amino acids, cysteine, and immunoglobulin (2).
There are different types of whey protein, including isolate,
concentrate, and hydrolysate, which can come in different for-
mulations including powder, milk, and specialized formula with
a higher content of certain amino acids. Compared to other
kinds of protein, whey protein is water-soluble, quickly digested,
and easily absorbed.
Theoretically, whey protein induces weight loss via
decreased appetite and increased satiety through several mech-
anisms such as regulation of satiety hormones, alteration of
hepatic gluconeogenesis, and diet-induced thermogenesis. To
improve body composition, whey protein positively stimulates
an anabolic effect because of a high concentration of branched-
chain amino acids and fast absorption. Of interest, whey pro-
tein has been found to lower blood pressure because of an
angiotensin-converting enzyme inhibitory property and aug-
mentation of nitric oxide–mediated vasodilation from the com-
ponent of isoleucine-proline-alanine tripeptide in whey
protein. Furthermore, whey protein consumption can improve
lipid metabolism by promoting lipoprotein lipase and inhibit-
ing cholesterol absorption.

CONTACT Anawin Sanguankeo, MD asangua1@jhmi.edu 1830 E. Monument St. 4th Floor, Suite 416, Baltimore, MD 21287, USA.
Color versions of one or more of the figures in the article can be found online at www.tandfonline.com/uacn.
© 2017 American College of Nutrition
 2 K. WIRUNSAWANYA ET AL.
Our study aimed to analyze all available published random-
ized controlled trials (RCTs) regarding the effect of whey pro-
tein supplementation on body composition including total
body weight, total fat mass, lean body mass, and also modifiable
cardiovascular risk factors in overweight and obese patients.
Methods
Materials and methods
This systematic review and meta-analysis were conducted and
reported according to the established guidelines for meta-
analyses of RCTs (4) and was registered in PROSPERO (regis-
tration number: CRD42016038344).
Search strategy
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Two authors (AS, SU) independently searched published stud-
ies indexed in MEDLINE, Embase, and the Cochrane Central
Register of Controlled Trials in the Cochrane Library. Referen-
ces of all selected studies were also examined. The following
main search terms were used: whey protein, body mass index
(BMI), body weight, lean body mass, fat-free mass, lean mass,
waist circumference, waist-hip ratio, body habitus, body fat,
insulin resistance, metabolic syndrome, overweight, and
obesity. The full search terms used are detailed in Appendix A.
Inclusion and exclusion criteria
Studies were selected in this meta-analysis if (a) study designs
were published RCTs; (b) whey protein isolate, concentrate, or
hydrolysate was the intervention; (c) participants were aged
18 years or older and were overweight or obese; (d) the study
had intervention of a minimum of 2 weeks; (e) the study had a
comparable placebo or control group; (f) body composition, fat
mass, and lean body mass were measured by dual-energy x-ray
absorptiometry (DEXA); and (g) the study assessed the out-
comes of body weight, body composition, insulin resistance,
metabolic syndrome, systolic blood pressure (SBP), diastolic
blood pressure (DBP), triglycerides, fasting blood glucose, low-
density lipoprotein (LDL) cholesterol, or high-density lipopro-
tein (HDL) cholesterol. Reviews, case reports, and abstracts
were excluded because their quality of studies could not be
assessed. Overweight and obesity are defined as BMI 25 and
30 kg/m2, respectively.
Data extraction
Two authors (AS, SU) independently reviewed titles and
abstracts of all citations that were identified. After all abstracts
were reviewed, data comparisons between the two investigators
were conducted to ensure completeness and reliability. The
inclusion criteria were independently applied to all identified
studies. Differing decisions were resolved by discussion
between the two authors.
Full-text versions of potentially relevant papers identified in
the initial screening were retrieved. If multiple articles from the
same study were found, only the article with more detailed
information was selected to avoid data duplication. Data
concerning authors, publication year, geographic location,
study design, source of information, baseline population char-
acteristics, type and dosage of whey protein, type of placebo or
control groups, and outcome assessment were independently
extracted. Articles in languages other than English were trans-
lated using Google Translate. We planned to contact the
authors of the primary reports to request any unpublished
data. If the authors did not reply, we used the available data for
our analyses.
Assessment of quality
The methodological quality of each study was assessed by two
authors (KW and SU) using the Cochrane Collaboration’s tool
for assessing risk of bias in randomized trials (5) on the follow-
ing items, in which each component was categorized as having
high, low, or unclear risk of bias: random sequence generation,
allocation concealment, blinding of participants and personnel,
blinding of outcome assessment, incomplete outcome data, and
selective reporting. Discrepant opinions between authors were
resolved by consensus.
Statistical analysis
We performed meta-analysis of the included studies using
Comprehensive Meta-Analysis 3.3 software from Biostat, Inc
(Englewood, NJ, USA). We calculated pooled mean difference
(MD) with 95% confidence intervals (CIs) comparing interven-
tion and control groups for each continuous outcome using a
random effects model because we hypothesized that there
would be high heterogeneity among studies. If only standard
deviation (SD) for the baseline and final values were provided,
the SD for the net changes were imputed according to the
method of Follmann et al. using a correlation coefficient of 0.5
(6). We excluded studies from meta-analysis if the outcome in
that study could not be combined with outcomes from other
studies. The heterogeneity of effect size estimates across these
studies was quantified using the Q statistic, its p value, and I2
(p < 0.10 was considered significant). The Q statistic compared
the observed between-study dispersion and expected dispersion
of the effect size and was expressed in p value for statistical sig-
nificance. An I2 is the ratio of true heterogeneity to total
observed variation. An I2 of 0% to 40% was considered to
exclude heterogeneity, 30% to 60% was considered to represent
moderate heterogeneity, 50% to 90% was considered to repre-
sent substantial heterogeneity, and 75% to 100% was consid-
ered to represent considerable heterogeneity (7). In the
presence of significant heterogeneity, subgroup analysis and a
meta-regression analysis was conducted to find the source of
heterogeneity (8). Publication bias was assessed using funnel
plot, Egger’s regression test, and its implications with the trim
and fill method (9).
Results
Description of included studies
The search identified 195 potentially eligible articles, of which
173 articles were excluded based on the title and abstract. A

total of 22 articles underwent full-length review. Thirteen
articles were excluded (4 articles were not observational studies,
3 articles had no control group, and 6 articles did not report
outcome of interest). Nine RCTs (10–18) involving 455 partici-
pants were included in the meta-analysis. Search methodology
is outlined in Appendix B. Characteristics of the included
studies are listed in Table 1. A summary of the risk of bias of
included studies is shown in Appendix C.
Our collected studies utilized varying dosages of whey pro-
tein from 20 g/d to 75 g/d during a time frame ranging from 2
weeks to 15 months. Different formulations of whey protein
had been utilized, such as normal powder, milk, and specialized
whey protein including a higher content of leucine and vitamin
D and lower fat content.n had been utilized, such as normal
powder, milk, and specialized whey protein including a higher
content of leucine and vitamin D and lower fat content”>
Some of our collected studies also restricted calorie intake to
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positively enhance the effectiveness of whey protein.
Whey protein and body composition
We performed the meta-analyses for body composition includ-
ing body fat (3 studies (13–15)), fat mass (3 studies (12,14,18)),
lean mass (4 studies (12–15)), body weight (6 studies
(11–13,15,17,18)), and waist circumference (5 studies
(11,12,15,17,18)). Comparing participants with whey protein
intake and controls, there were statistically significant differen-
ces in the changes of fat mass (MD D 1.12, 95% CI: 0.77–1.47,
I2 D 31%, pheterogeneity D 0.23), lean mass (MD D 0.77, 95% CI:
0.59–0.96, I2 D 0%, pheterogeneity D 0.43), and body weight (MD
D 0.56, 95% CI: 0.30–0.81, I2 D 0%, pheterogeneity D 0.64) favor-
ing the whey protein intake group, whereas there were no sig-
nificant differences in body fat (MD D ¡0.34, 95% CI: ¡1.15–
0.46, I2 D 0%, pheterogeneity D 0.74) and waist circumference
(MD D 0.46, 95% CI: ¡0.66–1.57, I2 D 84%, pheterogeneity D
1.27) between the whey protein group and controls. A forest
plot of comparison of body fat, fat mass, lean mass, and trunk
fat mass between the whey protein group and controls is shown
in Figure 1 and a comparison of body weight and waist circum-
ference is shown in Figure 2.
Whey protein and cardiovascular risk factors
We also performed a meta-analysis for cardiovascular risk fac-
tors including glucose (4 studies (10,15–17)), HDL cholesterol
(6 studies (10,12,13,15–17)), LDL cholesterol (6 studies
(12,13,15)), total cholesterol (6 studies (10,12,13,15–17)), and
triglycerides (6 studies (10,12,13,15–17)). Comparing partici-
pants with whey protein intake and controls, there were statisti-
cally significant changes favoring whey protein for glucose (MD
D 0.76, 95% CI: 0.14–1.38, I2 D 99%, pheterogeneity < 0.01), HDL
cholesterol (MD D 0.42, 95% CI: 0.05–0.80, I2 D 95%, pheterogeneity
< 0.01), and total cholesterol (MD D 0.53, 95% CI: 0.00–1.06,
I2 D 89%, pheterogeneity < 0.01), whereas there were no significant
differences in triglycerides (MD D ¡0.42, 95% CI: ¡0.79 to
¡0.06, I2 D 97%, pheterogeneity < 0.01) or LDL cholesterol (MD D
0.31, 95% CI: ¡0.02–0.63, I2 D 85%, pheterogeneity < 0.01) between
the whey protein group and controls. A forest plot of comparison
JOURNAL OF THE AMERICAN COLLEGE OF NUTRITION 3
of all cardiovascular risk factors in both groups is shown in
Appendix D.
Sensitivity analysis
We performed a meta-analysis for body composition and car-
diovascular risk factors including only studies that used DEXA
to measure body composition. Comparison of changes in fat
mass, lean mass, body weight, waist circumference, total choles-
terol, triglycerides, HDL cholesterol, and LDL cholesterol was
performed. Results for fat mass, lean mass, body weight, trigly-
cerides, HDL cholesterol, and LDL cholesterol were quite simi-
lar to those in the main analysis. There were statistically
significant changes favoring whey protein for waist circumfer-
ence (MD D 0.78, 95% CI: 0.38–1.19), but no significant differ-
ence in total cholesterol (MD D 2.42, 95% CI: ¡1.73–6.58).
Evaluation of publication bias
To investigate potential publication bias, we examined the con-
tour-enhanced funnel plot of the included studies that assessed
mean difference of body weight (Figure 3). The plot excludes
bias because there is symmetrical distribution of studies on
both sides of the mean. Furthermore, results of the Egger’s test
were nonsignificant (p D 0.97). Using the trim and fill methods
in the random effects model, there was no difference of the
imputed mean difference (MD D 0.58, 95% CI: 0.33–0.83).
Discussion
Obesity is one of the major contributing factors in cardiovascu-
lar disease and diabetes mellitus, and the management of obe-
sity is multifactorial and challenging (19–21). The primary
rationale of this meta-analysis was to provide a better under-
standing of the effect of whey protein on body composition
and cardiovascular risk factors among nondiabetic individuals
who are obese and overweight. The literature review revealed
that none of the previous meta-analyses have comprehensively
addressed the effect of whey protein on blood pressure. Our
meta-analysis demonstrated that the whey protein group shows
a statistically significant reduction in SBP, DBP, and other car-
diovascular risk factors including levels of fasting blood sugar,
total cholesterol, and HDL cholesterol compared to control
group. In addition, we found statistically significant improve-
ments in fat mass, lean body mass, and body weight in the
whey protein group compared to the control group. Therefore,
our results of this meta-analysis suggested that whey protein
supplementation could feasibly reduce some modifiable cardio-
vascular risk factors among nondiabetic individuals who are
obese and overweight.
According to our collected studies, most control groups
consumed a similar amount of protein compared to the
whey protein group, and this meta-analysis demonstrated
statistically significant reductions in SBP, DBP, HDL choles-
terol, total cholesterol, body weight, fat mass, and lean body
mass in the whey protein group. In addition, some of our
collected studies used a combination of whey protein con-
sumption and other modalities including a hypocaloric diet
and resistance exercise in the intervention group. However,
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Table 1. Characteristics of included studies.

K. WIRUNSAWANYA ET AL.

Demographics Whey Protein Group Control Group

Dosage of Other Form of Non– Body

Study and Study Participants BMI Form of Whey Whey Protein Whey Protein Dosage of Non– Composition
Year Duration Source Country (N) Age Female (kg/m2) Protein (g/d) Energy Supplement Whey Protein (g/d) Energy Measurement

Hambre, 2012 12 months Single research Sweden 24 24.2 0% 2a2.6 Protein powder 33 No restricted calories Fast food 41 1350 kcal/meal DEXA scan
center
Piccolo, 2015 8 weeks Multiple centers USA 27 41 § 9.8 100% 36.9 Whey-based protein 20 Energy-restricted diet Gelatin-based Gelatin: 20 Energy-restricted N/A
supplement supplement diet
Pal, 2010 (16) 3 weeks Single health Australia 20 <66 100% — Whey protein isolate 45 2589 kJ per meal Sodium casenate and Sodium casenate: 2589 kJ per meal N/A
clinic glucose 45
Figueroa, 2013 12 weeks Single center USA 33 30 § 1 100% 35.2 § 0.9 Milk protein 30 No restricted calories Casein and control Casein: 30 No restricted N/A
carbohydrate calories
Hector, 2015 2 weeks Multiple centers Canada 40 52 § 2 52.5% 34.7 § 1.1 Isolated whey protein 27 Hypoenergetic diet Soy and carbohydrate Soy: 26 Hypoenergetic diet DEXA scan
1750 § 123 kcal/d) (1760 § 142
kcal/d)
Verrigen, 2015 15 months Single center Netherlands 80 63 § 5.6 53% 33 § 4.4 Whey protein, leucine, 20 Hypocaloric diet of Placebo 0 Hypocaloric diet of DEXA scan
and vitamin 600 kcal below 600 kcal below
D–enriched estimated energy estimated energy
supplement in meal needs (2621 § 437 needs (2473 §
kcal/d) 636 kcal/d)
Frestedt, 2008 12 weeks Multiple centers USA 101 43.6 § 1.1 100% 35.8 Specialized whey fraction 75 1,700 cal/d Isocaloric beverage Not clear but lower 1,700 cal/d DEXA scan
(ProlibraTM , high in containing than
leucine, bioactive maltodextrin intervention
peptides, and milk group
calcium)
Pal, 2010 (15) 12 weeks Single center Australia 70 48.5 § 2.0 85.71% 31.3 § 0.8 Whey protein isolate 60 No restricted calories Casein and glucose Casein: 60 No restricted calories DEXA scan
Kinsey, 2014 2 weeks Multiple centers USA 44 27.4 § 5.0 100% 25.7 § 54.6 Whey protein isolate and 30 No restricted calories Casein and Casein: 30 No restricted calories DEXA scan
concentrate, powder carbohydrate
form placebo

Note. Data were presented as mean § standard deviation. All studies included were randomized controlled trials.
CMR D competitive meal replacement, BMI D body mass index; DEXA D dual-energy x-ray absorptiometry.
 JOURNAL OF THE AMERICAN COLLEGE OF NUTRITION 5

Figure 1. Forest plot of comparison of body fat, fat mass, lean mass, and trunk fat mass between whey protein intake group and controls. CI D confidence interval.
Downloaded by [Southern Cross University] at 08:03 02 November 2017

Figure 2. Forest plot of comparison of body weight and waist circumference between whey protein intake group and controls. CI D confidence interval.

five of our collected studies used only whey protein in the
intervention groups and they still demonstrated statistically
significant improvements on body composition and lipid
metabolism (10,13,15,16). It therefore could be suggested
that whey protein alone has a greater potential effect on
modifiable cardiovascular risk factors compared to non–
whey protein interventions regardless of use of other modali-
ties, including resistance exercise and a hypocaloric diet. A
recent randomized placebo-controlled trial by Fekete et al.
(22) suggested that whey protein consumption (56 g/d) for 8
weeks lowered SBP and DBP and improved lipid risk factors
and biomarkers of endothelial function compared to the con-
trol group. This study recruited 38 adults with prehyperten-
sion and mild hypertension and no restrict energy diet or
exercise was performed in this study. In the study by
Stojkovic et al. (23), improvement in body composition

Figure 3. Funnel plots showing publication bias in the studies reporting body weight of participants with whey protein intake and controls. Circles represent observed
published studies.
 6 K. WIRUNSAWANYA ET AL.
including reduction of BMI and fat mass and increased lean
body mass was found, and there was no statistically signifi-
cant changes in homeostatic model assessment–insulin resis-
tance (HOMA-IR). The results of our meta-analysis are in
agreement with that study. The study by Stojkovic et al. (23)
was conducted as a double-blind randomized trial for 48
weeks to investigate the potential benefits of whey protein on
body composition, HOMA-IR, and markers of inflammation
compared to maltodextrin supplementation. There was no
statistically significant change in HOMA-IR because the par-
ticipants in that study were not diabetic and not obese or
overweight and baseline HOMA-IR was relatively low
(<2%). Our study populations are different because we
included only obese and overweight patients. We found the
statistically significant reduction of fasting blood sugar but
we could not detect the reduction of HOMA-IR, which sug-
gested that whey protein may have some potential effect on
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glucose metabolism but a longer duration of study may have
been required to reveal significant changes in HOMA-IR.
Another previous study (24) investigating the effect of whey
protein consumption on postprandial blood sugar compared
to a high-carbohydrate diet found a statistically significant
decrease in postprandial glycemia in participants consuming
whey protein. The glycemic-lowering effect of whey protein
is thought to be due to incremental insulin levels after whey
protein consumption (24).
In our study, DEXA was conclusively chosen to be a method
for measuring body composition because of rapidness, avail-
ability, accuracy, inexpensiveness, and low radiation exposure
(25). DEXA has been widely used in many studies to measure
bone density and body composition, separating into fat mass
and free fat mass (25). We did not use BMI as a measurement
of body composition because BMI is no longer the best predic-
tor of cardiovascular risk factors and there are numerous stud-
ies suggesting that body composition provides a better
prognosis and prediction of cardiovascular risk factors
compared to BMI (61–64).
Body composition has been found to be strongly associ-
ated with cardiovascular and metabolic complications (26).
Because adipose tissue is the main organ of triglyceride stor-
age, excessive adipose tissue or increased fat mass can lead
to hypertriglyceridemia via lipolysis and increased lipid syn-
thesis in liver (27). A recent study has shown the correlation
of abdominal obesity and high liver fat content, which may
result in alteration of hepatic glucose output (26). Hence,
hyperglycemic state and insulin resistance are largely
explained by this phenomenon. Moreover, adipose tissue
functions as an endocrine gland secreting some chemokines
such as monocyte chemotactic protein-1, tumor necrosis fac-
tor-alpha, interleukin (IL) 1, IL-6, and IL-8, which could
contribute to a chronic inflammatory state and insulin resis-
tance (28). One article from the American Heart Association
demonstrated that a decrease in visceral adiposity was found
to ameliorate the cardiometabolic risk profile (26). Several
studies revealed that weight reduction by calorie restriction
and exercise and increased lean body mass are associated
with increased HDL cholesterol levels, which could be a pro-
tective factor for development of cardiovascular complica-
tions by antiatherogenic functions (29-31).
There are several plausible mechanisms of using whey
protein supplement to improve body composition. First,
increased levels of satiety hormones are the major mecha-
nism of improving body composition (12,32). Satiety hor-
mones including glucagon-like peptide-1 (GLP-1), dipeptidyl
peptidase 4, and cholecystokinin (CCK) are found to be
increased after whey protein consumption (32). CCK is a
peptide hormone that evidently inhibits gastric emptying
and induces satiety. The secretion of CCK into the gut is
effectively stimulated by protein ingestion and amino acids
(32–34). GLP-1 is secreted from enteroendocrine cells in the
gut, which is typically stimulated by an oral nutrient load.
GLP-1 inhibits gastric emptying and reduces insulin secre-
tion by blunting postprandial glucose response (35). Lipo-
genesis tends to be decreased because of the lower
hyperinsulinemia (35). Of interest, tripeptide Ile-Pro-Ala, the
hydrolytic breakdown of beta-lactoglobulin in whey protein,
has a similar effect as the dipeptidyl peptidase 4-inhibitor,
which can delay the degradation of GLP-1 (32,36,37). Sec-
ond, high-protein and low-carbohydrate diets have been
shown to be associated with positive alterations in hepatic
gluconeogenesis, which can increase plasma glucose levels,
prevent hypoglycemia, and possibly suppress appetite
(32,38,39). Third, diet-induced thermogenesis (DIT) could
feasibly play an important role in enhancement of weight
reduction with high-protein intake. Protein has been found
to have the highest DIT values compared to carbohydrates
and fats in diet. The increase in DIT may increase satiety by
the expansion of oxygen demand for protein metabolism,
which can potentially enhance satiety (32,40). Fourth, whey
protein also has an anabolic effect, which can stimulate mus-
cle synthesis more than other types of protein because of
faster absorption and a higher leucine component (32,41,42).
Besides the favorable effect on body composition, whey pro-
tein has been found to have favorable effects on lowering blood
pressure. The isoleucine-proline-alanine tripeptide in whey
protein may reduce blood pressure by an angiotensin-convert-
ing enzyme inhibitory property and by augmenting nitric
oxide–mediated vasodilation (43–45). Whey protein consump-
tion also has potential effects on some cardiovascular risk fac-
tors, including lipid metabolism. The underlying mechanisms
are stimulation of lipoprotein lipase, inhibition of cholesterol
absorption in the intestine mediated by beta-lactoglobulin and
sphingolipids in whey protein, and down-regulation of gene
expression in fatty acid transport and cholesterol absorption
from branched-chain amino acids (46,47). Of interest, several
studies revealed that weight reduction by calorie restriction and
exercise and increased lean body mass is associated with
increased HDL cholesterol levels with unclear underlying
mechanisms, which could be a protective factor for the devel-
opment of cardiovascular complications by antiatherogenic
functions (48). Some of our collected studies showed statisti-
cally significant benefits on lipid metabolism, and the results of
our meta-analysis are mostly consistent with their results.
However, we failed to detect reductions in triglycerides and
LDL cholesterol. The shorter durations of our collected studies
could limit the results.
Our study has several limitations. First and most notable,
the number of participants in the collected studies is low.

Given such small sample sizes, some studies may not have
been able to achieve statistically significant results. Thus, fur-
ther studies with broader sample sizes are highly recom-
mended. Second, the study durations were relatively short.
For instance, trunk fat mass, waist circumference, and
HOMA-IR are not likely to be affected in a 2- to 4-week
period of energy-restricted diet and whey protein supple-
mentation. Extended studies are necessary to elucidate the
prolonged effects on cardiovascular risk. Third, because
some aspects of the mechanism underlying whey protein’s
effect on human lipid metabolism are still poorly under-
stood, further investigation into the mechanisms of whey’s
effect on cholesterol response is recommended. Fourth, BMI
was used to determine obese and overweight status pre-inter-
vention and could be confounded by muscle mass. Last,
DEXA has some limitations including lack of specific cutoff
in individuals who are overweight and obese, under- or
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over-interpretation of the fat mass or lean mass from
changes in body hydration, and inaccuracy in estimation of
trunk composition in DEXA (49–51). More studies are war-
ranted to hopefully provide a standard reference value in
individuals who are overweight and obese.
In conclusion, our meta-analysis research reveals a num-
ber of potential favorable effects on cardiovascular risk fac-
tors such as body weight, fat mass, lean body mass, HDL
cholesterol, total cholesterol, and fasting glucose despite
some limitations. To the best of our knowledge, our study is
the first meta-analysis to examine the potential favorable
effects of whey protein supplementation on blood pressure
compared to non–whey protein supplementation. The
enhanced weight loss and improved cardiovascular risk asso-
ciated with this nutritional supplement is very encouraging
and should be investigated further for optimal regimen and
its long-term effect. We believe that whey protein supple-
mentation may be a legitimate alternative intervention to
improve cardiovascular risk in nondiabetic individuals who
are obese and overweight in the near future.
Acknowledgment
We thank Matthew Roslund for the validation of the search.
ORCID
Anawin Sanguankeo http://orcid.org/0000-0002-3662-4136
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 JOURNAL OF THE AMERICAN COLLEGE OF NUTRITION 9

Appendix A. Search strategy Embase

(((‘whey’/exp or whey) or (‘whey protein’/exp or ‘whey pro-
MEDLINE tein’)) and ((‘obesity’/exp or ‘obesity’) or overweight or obese) and
1. exp Whey/ (2176) ((‘body mass’/exp or ‘body mass’) or ‘body mass index’ or (‘body
2. whey proteins.mp. or exp Whey Proteins/ (12916) weight’/exp or ‘body weight’) or (‘lean body weight’/exp or ‘lean
3. body mass.mp. (184493) body weight’) or ‘lean body tissue’ or (‘fat free mass’/exp or ‘fat free
4. body mass index.mp. or exp Body Mass Index/ mass’) or ‘lean mass’ or (‘waist circumference’/exp or ‘waist cir-
(163605) cumference’) or (‘waist hip ratio’/exp or ‘waist hip ratio’) or ‘body
5. body weight.mp. or exp Body Weight/ (479290) habitus’ or (‘weight reduction’/exp or ‘weight reduction’) or
6. lean body mass.mp. (6107) (‘weight gain’/exp or ‘weight gain’) or (‘weight change’/exp or
7. lean body tissue.mp. (32) ‘weight change’) or (‘body fat’/exp or ‘body fat’) or (‘insulin resis-
8. fat-free mass.mp. (5614) tance’/exp or ‘insulin resistance’) or homa or (‘metabolic syndrome
9. lean mass.mp. (3270) x’/exp or ‘metabolic syndrome x’) or (‘cardiovascular disease’/exp
10. waist circumference.mp. or exp Waist Circumference/ or ‘cardiovascular disease’))) and [embase]/lim not [medline]/lim
(18953) CENTRAL
11. waist to hip.mp. (10327) #1 MeSH descriptor: [Whey] explode all trees 261
12. Waist-Hip Ratio/(3151) #2 MeSH descriptor: [Whey Proteins] explode all trees 461
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13. body habitus.mp. (1078) #3 MeSH descriptor: [Body Mass Index] explode all
14. weight loss.mp. or exp Weight Loss/ (75889) trees 7501
15. weight gain.mp. or exp Weight Gain/ (58373) #4 MeSH descriptor: [Body Weight] explode all trees 18906
16. weight change.mp. (5244) #5 lean body mass 2243
17. body fat.mp. or exp Fat Body/ (26421) #6 fat-free mass 1074
18. Insulin Resistance.mp. or exp Insulin Resistance/ #7 lean mass 2329
(86927) #8 MeSH descriptor: [Waist Circumference] explode all
19. HOMA.mp. (10534) trees 626
20. exp Metabolic Syndrome X/or Metabolic Syndrome.mp. #9 MeSH descriptor: [Weight Loss] explode all trees 4301
(38782) #10 MeSH descriptor: [Weight Gain] explode all trees 1872
21. exp Obesity/or obesity.mp. (228902) #11 MeSH descriptor: [Fat Body] explode all trees 1
22. overweight.mp. or exp Overweight/ (179440) #12 MeSH descriptor: [Insulin Resistance] explode all
23. Cardiovascular Diseases.mp. or exp Cardiovascular trees 3802
Diseases/ (2041755) #13 MeSH descriptor: [Metabolic Syndrome X] explode all
24. 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 or 11 or 12 or 13 or trees 1205
14 or 15 or 16 or 17 (645173) #14 MeSH descriptor: [Obesity] explode all trees 9168
25. 18 or 19 or 20 or 23 (2117058) #15 MeSH descriptor: [Overweight] explode all trees 10066
26. 21 or 22 (243388) #16 obese 11089
27. 24 or 25 (2643675) #17 #1 or #2 462
28. 1 or 2 (12934) #18 #3 or #4 or #5 or #6 or #7 or #8 or #9 or #10 or #11 or
29. obese.mp. (92772) #12 or #13 26374
30. 21 or 22 or 29 (260268) #19 #14 or #15 or #16 15514
31. 27 and 28 and 30 (86) #20 #17 and #18 and #19 35
 10 K. WIRUNSAWANYA ET AL.

Appendix B. Search methodology and selection process

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Appendix C. Summary of risk of bias among included studies. Positive sign means low risk of bias. Negative
sign means high risk of bias
 JOURNAL OF THE AMERICAN COLLEGE OF NUTRITION 11

Appendix D. Forest plot of comparison of glucose, total cholesterol, triglyceride, high-density lipoprotein
(HDL) cholesterol, and low-density lipoprotein (LDL) cholesterol between the whey protein intake
group and controls. CI D confidence interval
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