Effect of the β3-adrenergic receptor gene polymorphism Trp64Arg on BMI reduction associated with an exercise-based intervention program in Japanese middle-aged males
Environ Health Prev Med
Effect of the b3-adrenergic receptor gene polymorphism Trp64Arg on BMI reduction associated with an exercise-based intervention program in Japanese middle-aged males
Aya Tahara 0
Yoneatsu Osaki 0
Takuji Kishimoto 0
0 A. Tahara (&) Y. Osaki T. Kishimoto Division of Environmental and Preventive Medicine, Department of Social Medicine, Faculty of Medicine, Tottori University , Tottori 683-8503 , Japan
Objectives The b3-Adrenergic receptor gene polymorphism Trp64Arg (ADRB3 Trp64Arg) may be associated with weight gain, especially in the East Asian populations. To date, however, the results of various investigations aimed at determining whether ADRB3 Trp64Arg has any effect on weight reduction after intervention have been inconsistent. The aim of this study was to assess the effect of ADRB3 Trp64Arg on weight reduction in a non-pharmacological intervention program. Methods Fifty-seven Japanese men (average age 48.1 ± 5.9 years) whose body mass index (BMI) was [23.0 kg/m2 participated in an intervention program in which they were encouraged to exercise by walking with a pedometer for 12 weeks. The BMI was measured at the start of the intervention program and on weeks 4, 8, and 12 (end) of the intervention period. The reduction in the BMI (DBMI) was calculated as the BMI value obtained at each point in comparison to the baseline (initial) BMI. The participants were classified into two groups: Arg allele non-carriers (Trp/Trp) and Arg allele carriers (Trp/Arg or Arg/Arg), respectively. Results Among the participants, 61.4% were Trp/Trp homozygous, 38.6% were Trp/Arg heterozygous, and 0.0% were Arg/Arg homozygous. There was no significant difference in the DBMI between Arg allele non-carriers and Arg allele carriers at week 4 [non-carrier mean ± standard deviation (SD) vs. carrier mean ± SD: 0.25 ± 0.43 vs. 0.24 ± 0.35, respectively; p = 0.954], week 8 (0.56 ± 0.68 vs. 0.58 ± 0.61, respectively; p = 0.913), and week 12 (0.70 ± 0.86 vs. 0.84 ± 0.80, respectively; p = 0.545). Conclusions Our results suggest that ADRB3 Trp64Arg does not influence BMI reduction after an exercise-based intervention program.
Exercise; Interventional study; Middle-aged men; Trp64Arg; Weight reduction
Overweight and obese individuals have an increased risk of
coronary disease, stroke, and some cancers. The number of
such individuals is increasing globally [
], and in the USA
these increasing numbers have led to the prediction of a
shortened average life span [
]. Physical activity and
improved dietary habits are effective strategies to reduce
body weight, and they can also decrease the risk of
associated cardiovascular disease and stroke [
there are individual differences in the effects of such
intervention programs, even among individuals following
the same weight reduction program. Genetic factors are
associated with individual differences in weight gain [
and it is possible that differences in weight reduction may
also be caused by hereditary factors.
A number of different genes are believed to be
associated with human obesity [
]. b3-Adrenergic receptors,
which are mainly expressed in white and brown adipocytes,
control lipolysis and heat production induced by
]. A polymorphism resulting in a switch from
tryptophan to arginine at the 64th residue in the
b3adrenergic receptor gene, known as the ADRB3 Trp64Arg,
was identified in 1995 [
]. The results of many
subsequent studies suggest that ADRB3 Trp64Arg is
associated with weight gain, especially in East Asian
], thereby also leading to the possibility
that ADRB3 Trp64Arg also influences weight reduction.
Several studies have compared weight reduction in
individuals with the different ADRB3 Trp64Arg genotypes
participating in non-pharmacological intervention
]. Studies in Korea and the USA reported that
there was no difference in weight reduction between Arg
allele carriers and non-carriers [
]. In contrast,
findings of Japanese studies have been inconsistent, with five
studies showing a significant difference associated with
ADRB3 Trp64Arg [
] and one study showing no
]. It has been suggested that the differences
observed in the results of the former five Japanese studies
were caused by differences in the race of a subject, sex,
weight at baseline, and/or the amount of weight loss by
intervention. Most intervention studies have been carried
out using female subjects, and there are little available data
at the present time on Japanese males. Therefore, the aim
of the study reported here was to examine the effect of
ADRB3 Trp64Arg on weight loss associated with a
nonpharmacological intervention program for Japanese male
Materials and methods
Ninety-three Japanese workers were recruited to participate
in the intervention program for 12 weeks. The program
was based mainly on walking. The inclusion criteria of this
study were male gender, body mass index (BMI) C23.0 kg/
], and no past history of cardiovascular disease,
cancer, or renal disease. Sixty-six men met these criteria,
among whom nine were subsequently excluded because
they did not provide data during business trips. Thus, data
on 57 men were included in the final analysis. The average
age of the subjects was 48.1 ± 5.9 years. The study
protocol was reviewed and approved by the Ethics Committee
of Tottori University School of Medicine. All of the
participants provided written informed consent to participate
in this study.
The participants participated in an intervention program for
12 weeks during which they wore a pedometer Kenz
Lifecorder EX (Suzuken, Tokyo) and started walking. The
goal was 10,000 steps per day. The accumulated number of
steps walked was checked once a week. The subjects
received individual nutritional guidance from a dietitian at
least once during the 12-week period. They also
participated in supportive group therapy aimed at providing
information on arterial sclerosis and at reviewing their
efforts at the initiation of their intervention program, week
6, and at the end of the program. Both the exercise habits
and eating habits before and after the intervention program
were assessed by a self-administered questionnaire.
Measurement of anthropometric parameters
The subjects’ height was measured at the start of the
intervention. Their body weight and waist circumference
were measured once a week from the start of the
intervention until it was concluded. Participants wore only light
clothing at the time of the body weight measurement, and
the 0.5 kg allotted for the weight of clothes was subtracted
from the value shown on the scale. The BMI was
calculated as the weight (in kilograms) divided by the squared
height (in square meters). The waist circumference was
measured to the nearest 0.1 cm at the umbilicus level, with
the subjects standing erect, abdomen relaxed, arms at the
sides, and the feet together. The reduction (D4th, D8th,
D12th week) was defined as: (the value after
intervention - the value before intervention).
Identification of ADRB3 Trp64Arg
The Trp64Arg variation of the b3-adrenoceptor gene was
examined in peripheral blood leukocyte DNA using a
TaqMan assay (Roche Diagnostics, Branchburg, NJ). The
DNA was extracted using the QIAamp DNA Blood Mini
Kit (Qiagen, Tokyo, Japan) according to the
manufacturer’s protocol. Genomic DNA was amplified using the
TaqMan SNP Genotyping Assay containing the VIC probe
and FAM probe (assay ID: C_2215549_20) and the
TaqMan Universal PCR Master Mix containing AmpliTaq
Gold DNA polymerase according to the following
thermocycle profile: initial incubation steps of 10 min at 95 C,
followed by 40 cycles of 15 s at 92 C and 1 min at 60 C.
Genotyping was conducted by detecting fluorescent signals
of allele-specific probes on ABI PRISM 7900HT Sequence
Detection System v2.1 (Applied Biosystems, Foster, CA).
The participants were notified of their genotype at the end
of the intervention period upon request.
The v2 test was used to determine the Hardy–Weinberg
equilibrium of the genotype frequency. The participants
were divided into two groups: non-carriers of the Arg allele
(Trp/Trp) and carriers of the Arg allele (Trp/Arg or Arg/
Arg), respectively. Twenty-seven people in each group
were necessary to estimate the indispensable sample size
based on a Japanese report [
], although the study
detected a power of 90%, a significance level of 5%, and a
difference in BMI reduction and its standard deviation
(SD) between the two groups of 0.17 and 0.19 kg/m2 [
In addition, one group needed 34 people since the other
was limited to 22 individuals. Reduction of anthropometric
parameters by intervention was defined as D and calculated
as: D4th week = (baseline value - 4th week value); D8th
week = (baseline value - 8th week value); D12th week =
(baseline value - 12th week value). Comparison of
frequencies among category variables was performed by v2 or
the Fisher’s exact test, where appropriate. Student’s t test
was used to compare the difference in anthropometric
parameters and its reduction between the two groups. In
terms of the efficacy of the intervention, each weekly value
was compared with the baseline by the paired t test.
Quantitative variables were expressed as the mean ± SD. For the
purpose of adjusting for potential confounding factors,
such as age, number of steps, and dietary modification
during the intervention, we adopted an analysis of
covariance to compare the intervention effect between the two
groups. Statistical analysis was conducted using the SPSS
ver. 15.0 statistical software package (SPSS, Chicago, IL).
A p \ 0.05 was considered to be statistically significant.
Thirty-five individuals (61.4%) had the Trp64/Trp64
genotype (Arg allele non-carriers), while 22 (38.6%) had
Number of participants
Age, years (mean ± SD)
Current smoker, n (%)
Everyday drinker, n (%)
Habitual exercisers, n (%)
Modifications in eating habits,
Taking medicines, n (%)
Height, cm (mean ± SD)
Body weight, kg (mean ± SD)
BMI, kg/m2 (mean ± SD)
Waist circumference, cm
(mean ± SD)
Number of steps/day (mean ± SD)
Modifications in eating habits, n (%)
the Trp64/Arg64 genotype (Arg allele carriers). No
participant was a carrier of the Arg64/Arg64 genotype. The
allele frequency in ADRB3 Trp64Arg was 92 for the Trp
allele (80.7%) and 22 for the Arg allele (19.3%), which
confirmed the Hardy–Weinberg equilibrium (p = 0.071).
A comparison of characteristics between Arg allele
noncarriers and carriers is shown in Table 1. There was no
significant difference between the two groups in terms of
age, current smoking, everyday alcohol consumption,
habitual exercisers, modifications in eating habits,
medication of anti-hypercholesterolemic drugs, diabetic drugs,
height, body weight, BMI, and waist circumference at
baseline. There was a difference in number of steps and
awareness of eating habits during the intervention between
two groups. Table 2 shows the initial values at baseline and
the changes in anthropometric parameters at each time
point during the intervention according to ADRB3 gene
variation. All anthropometric parameters, with the
exception of waist circumference at week 4, improved
significantly in both groups. The reduction from baseline grew
larger weekly during the intervention period in both
groups. We observed no significant difference in the
reduction of body weight, BMI, and waist circumference
between Arg allele non-carriers and Arg allele carriers at
weeks 4, 8, and 12. However, the difference in the
improvement of BMI between two groups was greater in
the 12th week than in the 4th and 8th weeks. After
adjusting for potential confounding factors, we found no
significant difference in the reduction in body weight, BMI,
Arg allele non-carriers
Arg allele carriersa
Data are given as the mean ± SD
a p value with Student’s t test between Arg allele non-carrier and Arg allele carrier at each point
b p values were calculated by analysis of covariance adjusting for age, number of steps during intervention, modifications in eating habits during
intervention, and observation time
c p \ 0.01 with the paired t test between baseline and each week
and waist circumference between the two groups. The
difference of the intervention effect on the anthropometric
parameters between those participants on medication and
those without was compared (data not shown). There was
no significant difference in the values before intervention
and the reduction during intervention in terms of all
anthropometric parameters between the participants on
anti-hypercholesterolemic or diabetic drugs and those not
receiving such medications (data not shown).
The aim of this study was to evaluate the difference in BMI
reduction at 4-week intervals between Japanese
middleaged men who were either Arg allele non-carriers of
ADRB3 Trp64Arg or Arg allele carriers and who were
participating in an intervention program. No difference was
observed at weeks 4, 8, and 12, respectively, of the
intervention program between non-carriers and carriers in terms
of BMI reduction from the baseline. In other words, the
BMI reduction during an exercise-based intervention was
not affected by the ADRB3 Trp64Arg polymorphism.
Among our subjects, the frequency of the Arg allele was
19.3%, which is similar to frequencies (16.6–25.0%)
reported among Japanese in other studies [
Kuriyama et al. [
] reported that ADRB3 Trp64Arg did
not affect weight reduction in a 6-month diet- and
exercisebased intervention for middle-aged Japanese individuals.
Although there was a difference in the sex ratio and in the
average age of the participants (approximately 15 years) in
the two groups (carriers and non-carriers), the results of their
study are consistent with our results. In contrast, Shiwaku
et al. [
] showed that Japanese peri-menopausal females
who were Arg allele carriers had difficulty obtaining optimal
results when participating in a weight reduction diet- and
exercise-based intervention program. In their study, the BMI
reduction after a 3-month intervention was approximately
0.2 kg/m2, which was smaller than that at the fourth week of
our study. Although the BMI had fallen by 0.8–0.9 kg/m2 by
the third month of our intervention, there is a possibility that
a less stringent intervention, such as a 0.2 kg/m2 of BMI
reduction, may have revealed a difference between the Arg
allele non-carriers and carriers. In their studies on Japanese
obese females, Yoshida et al. [
], Sakane et al. [
et al. [
], and Lee et al. [
] reported that Arg allele carriers
had difficulty losing weight. The BMI reduction in these
studies was larger, about 2 kg/m2 in Arg allele non-carriers
and 3 kg/m2 in Arg allele carriers. Possible factors
accounting for this inconsistency in the results of these
studies include sex, obesity level at baseline, and the
intensity of intervention. Previous BMI reduction studies
associated with ADRB3 Trp64Arg in Korea and the USA
found no association between ADRB3 Trp64Arg and weight
reduction; these findings are consistent with those of our
Our study had a number of limitations. First, the
inability to recruit a larger number of individuals for the
interventional study design resulted in a relatively small
sample size. If the difference in the BMI reduction between
non-carriers and carriers had been smaller, then the number
of study subjects was insufficient to detect an influence of
ADRB3 Trp64Arg on BMI reduction. Second, based on
our results, no conclusion can be reached on the effect of
the homozygous Arg64 allele on weight reduction. Oizumi
et al. [
] compared Trp64/Trp64 homozygous carriers
with others in a cross-sectional study and showed that the
BMI of homozygous Arg64/Arg64 carriers was larger than
that of homozygous Trp64/Trp64, with no difference with
the heterozygous Trp64/Arg64 carriers. An increased
sample size would be required to examine the influence of
homozygous Arg64/Arg64 on weight reduction because the
frequency of this genotype is low [
]. Third, the
intervention period only lasted 12 weeks. Although the
reductions in the BMI at the 4th and 8th weeks between the two
groups were similar, the reduction in the BMI at the 12th
week in Arg allele carriers was greater (but not
significantly greater) than that in Arg allele non-carriers.
Therefore, if the intervention period had continued for an even
longer period, then a difference according to ADRB3
polymorphism may have been detected. Therefore, future
intervention studies require longer observational periods.
Fourth, the influence of improved eating habits during the
intervention may not have been sufficiently taken into
consideration. The improvement in eating habits tended to
differ from person to person, and it was difficult to control
such dietary modification. Due to this situation, statistic
adjustment according to dietary modification/no dietary
modification during the intervention was performed.
In conclusion, we did not observe any association
between the decreasing pattern of the BMI and the ADRB3
Trp64Arg during an exercise-based intervention program
for middle-aged Japanese males with a BMI [23.0 kg/m2.
These results suggest that ADRB3 Trp64Arg does not
influence any reduction in the BMI during an
exercisebased intervention program.
Acknowledgments This study was supported by a Grant-in Aid for
Scientific Research (B) (term of project, 2006–2009; project number,
18390180) from the Japan Society for the Promotion of Science, Japan.
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