P.

Wang1,2
L. H. Ma1
H. Y.Wang3
W. Zhang1
Q. Tian1
Association between Polymorphisms of
D. N. Cao1
Vitamin D Receptor Gene ApaI, BsmI and TaqI and G. X. Zheng1
Muscular Strength in Young Chinese Women Y. L. Sun1
Physiology & Biochemistry

Abstract a Hardy-Weinberg equilibrium. Subjects with the vitamin D re-

The association between the polymorphisms of vitamin D recep-
tor gene ApaI, BsmI and TaqI and isokinetic muscular strength
was examined in 109 healthy, young Chinese women. Methods:
Genomic DNA was extracted from mouthwash samples and vita-
min D receptor genotypes were determined with the established
ceptor ApaI null (A allele) mutation (AA) exhibited significantly
lower knee and elbow concentric or eccentric peak torque than
those with aa homozygous or Aa heterozygous. The BB & Bb
group showed significantly higher peak torque in concentric
knee flexors than bb group. No association was found between
vitamin D receptor gene TaqI and muscular strength. The data in-

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methods. The peak torque of the dominant limb in concentric
and eccentric knee extensors and flexors and elbow extensors
and flexors was measured using Cybex-Norm-770 isokinetic dy-
namometer at slow and fast velocities. The distribution fre-
quency of vitamin D receptor gene alleles was analyzed by chi-
square test. The difference in muscular strength between VDR
genotype groups was analyzed using one-way ANOVA. The ApaI,
BsmI and TaqI alleles and genotype frequencies appeared to be in
dicated that vitamin D receptor gene ApaI and BsmI polymor-
phisms rather than TaqI polymorphism might be associated with
muscular strength.
Key words
Isokinetic peak torque · knee/elbow extensors · knee/elbow flex-
ors

182
Introduction
Recent studies on human genetics and heredity have suggested
that the variation in muscular strength might be attributed to
hereditary factors to some extent [19]. To date, however, little
evidence is available on specific genetic markers that may be
associated with human muscular strength.
Vitamin D receptor (VDR) is a critical mediator in gene transcrip-
tion. It has been found to be involved in genomic and non-ge-
nomic actions in vitamin D metabolism in skeletal muscle tissue
[1, 2,10]. There is a VDR-dependent non-genomic mechanism
which mediated 1.25-dihydroxyvitamin D affecting the activa-
tion of tyrosine phosphorylation pathways, which may contrib-
ute to hormone regulation of muscle growth [3]. Other studies
have also demonstrated that VDR plays an important role in
stimulating muscle cell proliferation and differentiation via
VDR –/– mice [5].
There have been studies that utilized polymorphisms in intron 8
(B/b and A/a alleles, at the BsmI and ApaI sites) and exon 9 (T/t
alleles, at the TaqI site) of vitamin D receptor gene in association
with some diseases [9,12,15]. Mirrison et al. have investigated
the relationship between VDR gene BsmI, ApaI and EcoRV poly-

Affiliation
1
Tianjin Research Institute of Sports Medicine, Tianjin, P. R. China
2
Beijing Genomics Institute Chinese Academy of Science, Beijing, P. R. China
3
Tianjin University of Science & Technology, Tianjin, P. R. China

Correspondence
Wang Pei · Beijing Airport Industrial Zone B-6 · Shun Yi district · Beijing · P. R. China 101300 ·
E-mail: wangpei7616@yahoo.com.cn

Accepted after revision: March 9, 2005

Bibliography
Int J Sports Med 2006; 27: 182 – 186 © Georg Thieme Verlag KG · Stuttgart · New York ·
DOI 10.1055/s-2005-865626 · Published online August 30, 2005 ·
ISSN 0172-4622
morphisms and BMD (Body Mineral Density), that opened up a
new way of examining the function of VDR gene [15]. Ferrari et
al. have demonstrated that the change of lumber spine BMD is
associated with the genotype of VDR gene, BsmI [7]. Matsuyama
has further reported that the more common genotypes in the
Japanese population of bbaaTT and bbAaTT are associated with
a positive response to 1α-hydoxy vitamin D3 therapy. In con-
trast, poor response has been reported in the genotypes of
BbAaTt, which is the most common in Caucasians [14]. Further-
more, there have been several reports on an association of gene
polymorphisms and muscle strength [17,18, 20]. Geusens et al.
have reported that VDR (BsmI) genotype is associated with the
strength of the quadriceps muscle in non-obese elderly women
[8].
Based on those studies, it was speculated that VDR gene might be
a candidate genetic marker for human muscular strength. To our
knowledge, there has been no report on the association of VDR
alleles and muscular strength in isokinetic contractions in pre-
menopausal women. The aim of this study was to examine
whether a relationship existed between the ApaI and BsmI and
TaqI polymorphisms at VDR gene loci and isokinetic muscular
strength in healthy, young Chinese women.
Materials and Methods
Subjects
One hundred and nine (109) women of Han nationality were ran-
domly recruited from the university student population in Tian-
jin, China. All subjects were healthy and without cardiovascular
or musculoskeletal conditions that might be aggravated by
strength testing, and all completed a physical activity question-
naire. It was found that the subjects had a similar level of daily
physical activity and none of them had previously participated
in weight training exercise. The subjects were given a thorough
explanation of the experimental procedures and signed a written
informed consent. The mean (± SD) age, body mass and height of
the subjects are reported in Tables 1 – 3.
Muscular strength
The strength tests were performed on a Cybex-Norm-770 isoki-
netic dynamometer. The maximal voluntary concentric (Con)
and eccentric (Ecc) peak torque were assessed for the dominant
leg. The peak torque of knee extensors (KE) and flexors (KF) was
recorded at velocities of 308/s, 1208/s and 1808/s, respectively.
Table 1 Association analysis of anthropometric phenotypes and VDR gene ApaI genotypes
Index Genotype
AA Aa
n 7 47
Age (yr) 19.57 ± 0.53 20.00 ± 1.20
Ht (cm) 155.66 ± 5.26 160.09 ± 5.77
Wt (kg) 49.66 ± 7.98 55.40 ± 7.70
Ht: height; Wt: weight; P1: AA vs. Aa; P2: AA vs. aa; P3: Aa vs. aa
Wang P et al. VDR Gene and Muscular Strength In Chinese … Int J Sports Med 2006; 27: 182 – 186
The peak torque of the dominant elbow extensors (EE) and flex-
ors (EF) was also measured in concentric contractions at veloc-
ities of 308/s and 1208/s, respectively. For each test, subjects per-
formed three maximal contractions, separated by a 30-s recovery
period in concentric tests or 90-s in eccentric tests. The highest
value of the three trials was accepted as the peak torque. Reliabil-
ity of the strength test was carried out prior to the formal mea-
surement. Intraclass correlation coefficients for the test-retest
reliability ranged from 0.89 to 0.99 for the knee and elbow tests.
Genotype
Genomic DNA was extracted from mouthwash samples [21]. VDR
genotypes of TaqI, ApaI and BsmI polymorphisms were deter-
mined based on the protocol outlined previously [9,16]. Subjects
Physiology & Biochemistry
were categorized by the exhibition of the TT, Tt, or tt genotype at
TaqI site; the aa, Aa or AA genotype at ApaI site; and the bb, Bb or
BB genotype at BsmI site.
Statistic
Differences between the VDR TaqI, ApaI and BsmI gene allele and
genotype distributions were assessed by a χ2 analysis. One-way
ANOVA was used to analyze the differences in physical character-
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istics between VDR genotypes. The results were reported as
means ± SE. Statistical significance was accepted at p ≤ 0.05.
Results
There were no significant differences in any physical characteris-
tics between the VDR TaqI and BsmI genotype groups. The body
height of the AA group was significantly lower than that of the aa
group, but no significant difference was found in other physical
characteristics (Tables 1 – 3). Of the 109 subjects in the study, 183
101 exhibited TT genotype, 7 exhibited the Tt genotype, and 1 ex-
hibited the rare tt genotype. The T allele frequency was 95.87 %,
and the t allele frequency was 4.13 %. Also, 55 subjects exhibited
the aa genotype, 47 exhibited the Aa genotype and 7 exhibited
the AA genotype. The allele frequency was 72.02 % and the A al-
lele frequency was 27.98 %. Only 96 subjects demonstrated the
BsmI site polymorphism. Of the 96 subjects, 85 exhibited the bb
genotype, 10 exhibited the Bb genotype and 1 exhibited the rare
BB genotype. The b allele frequency was 93.75 % and the B allele
frequency was 6.25 %. All of these values did not differ signifi-
cantly from predicted Hardy-Weinberg equilibrium (χ2 analysis,
p > 0.05).
P
aa P1 P2 P3
55
19.89 ± 1.05 0.336 0.469 0.617
160.65 ± 5.44 0.052 0.028 0.619
54.77 ± 8.25 0.079 0.114 0.690
 Table 2 Association analysis of anthropometric phenotypes and Table 3 Association analysis of anthropometric phenotypes and
VDR gene BsmI genotypes VDR gene TaqI genotypes

Index Genotype p Index Genotype p

BB & Bb bb TT Tt & tt

n 11 85 n 101 8
Age (yr) 20.18 ± 1.72 20.02 ± 1.18 0.779 Age (yr) 19.91 ± 0.95 20.00 ± 2.33 0.633
Ht (cm) 161.24 ± 7.10 160.28 ± 5.69 0.674 Ht (cm) 160.12 ± 5.67 162.43 ± 5.78 0.294
Wt (kg) 54.15 ± 4.58 53.91 ± 7.68 0.379 Wt (kg) 54.53 ± 7.85 56.08 ± 8.41 0.459

Ht: height; Wt: weight Ht: height; Wt: weight

Physiology & Biochemistry

Table 4 Association analysis of KF& KE Ecc PT and VDR gene ApaI genotypes

Index Genotype P
AA Aa aa P1 P2 P3

n 5 28 32

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1208/s KF Ecc, Nm 60.00 ± 10.20 68.79 ± 14.06 66.84 ± 11.00 0.149 0.255 0.547
308/s KF Ecc, Nm 60.80 ± 14.13 67.71 ± 12.28 66.53 ± 12.12 0.252 0.337 0.712
1208/s KE Ecc, Nm 90.20 ± 18.95 125.64 ± 24.92 126.69 ± 24.22 0.004 0.003 0.870
308/s KE Ecc, Nm 116.60 ± 26.31 138.68 ± 28.37 139.31 ± 25.73 0.097 0.085 0.928

P1: AA vs. Aa; P2: AA vs. aa; P3: Aa vs. aa

Table 5 Association analysis of EF Con PT and VDR gene ApaI genotypes

184
Index Genotype P1 P2
AA Aa aa

n 5 45 51
1208/s EF Con, Nm 11.80 ± 2.17 14.31 ± 2.28 15.20 ± 4.01 0.107 0.029
308/s EF Con, Nm 14.40 ± 2.30 17.84 ± 2.84 19.08 ± 6.09 0.129 0.039

P1: AA vs. Aa; P2: AA vs. aa

Since there was only one sample in tt and BB genotype groups
respectively, it was impossible to conduct statistical analyses on
these cases. Thus, they were combined with the Tt and Bb groups
respectively and the Wilcoxon Rank Sum method was applied to
examine the significance between the bb and the Bb&BB com-
bined groups, as well as between the TT and the Tt&tt combined
groups. ANOVA was utilized in analyzing the muscular strength
difference in ApaI genotypes.
With respect to the ApaI polymorphism site, the peak torque of
the dominant leg in KE Ecc at 1208/s in the AA homozygous group
was significantly lower than that of the aa and Aa groups
(p = 0.003, p = 0.004, respectively). The peak torque of the domi-
nant leg in KE Ecc at 308/s in the AA homozygous group was also
lower than that of other groups, but the difference was not signi-
ficant (p = 0.085, p = 0.097) (Table 4). The peak torque of the
dominant leg in EE and EF Con at both 1208/s and 308/s in the
AA homozygous group was significantly lower than that of the
aa homozygous group (p = 0.029, p = 0.039, respectively) (Table
5). In the BsmI polymorphism site, the peak torque of the domi-
nant KF Con at 1808/s in the Bb & BB combined group was signifi-
cantly higher than that of the bb homozygous group (p = 0.030).
The Bb&BB combined group also exhibited higher KF Con peak
torques at 1208/s and 308/s, but the difference was not statisti-
cally significant (Table 6). There was no significant difference be-
tween VDR gene TaqI genotypes in muscular strength.
Discussion
There have been several investigations into specific genetic
markers associated with human physical performance and mus-
cular strength. It had been reported that the heritability of
muscle isokinetic contraction and muscle explosive strength

Wang P et al. VDR Gene and Muscular Strength In Chinese … Int J Sports Med 2006; 27: 182 – 186

Table 6 Association analysis of KF Con PT and VDR gene BsmI geno-
types
Index Genotype
BB & Bb bb
n 11 83
1808/s KF Con, Nm 34.45 ± 5.16 30.68 ± 7.03
1208/s KF Con, Nm 38.36 ± 7.31 35.82 ± 7.49
308/s KF Con, Nm 55.91 ± 6.16 53.19 ± 12.01
KF: Con 1808/s peak torque value was only available for 80 subjects for bb genotype.
was nearly 0.83 – 0.97 [6,13]. Our result showed that the Aa and
aa genotype groups and the BB & Bb combined group showed
significantly higher muscle isokinetic strength and the most sig-
nificant differences were at fast isokinetic contractions. To our
knowledge, no similar study has been published before in the lit-
erature available to us. Meanwhile, in contrast to the distribu-
tions of VDR polymorphisms and association studies in Cauca-
sians, the present results suggest a racial difference in the distri-
bution of VDR alleles as well as the relationship between VDR ge-
netic polymorphisms and the phenotypes of muscle strength.
The Han population used in this study demonstrated a signifi-
cant predominance of KF isokinetic peak torque in the BB & Bb
combined group.
Up to now, the molecular basis of these polymorphisms in the
VDR gene is still not clear. The ApaI and BsmI are located at intron
8, so they might not cause the null protein. But their mutations
could affect transcription of the VDR gene. Carling et al. have
shown that the B alleles are associated with higher VDR mRNA
levels in a study on patients with sporadic primary hyperpara-
thyroidism [4]. The TaqI is located at the encoding segment of
VDR gene (9th exon of VDR gene.). However, Haussler et al. re-
ported that the mutation of TaqI could not debilitate the compo-
sition of VDR protein, so it might not impair the VDR function
[11]. Mirrison et al. demonstrated that the alleles were in strong
linkage disequilibrium with other polymorphisms. With respect
to kidney and bone cells, luciferase reporter gene activity from
the Bat haplotype was substantially greater than from the baT
haplotype, which is consistent with the enhanced gene tran-
scription or mRNA stability [15]. Although the results from sev-
eral cohort studies demonstrated the existence of an association
between the VDR gene alleles and muscular strength, the mecha-
nisms that influence muscle strength need further investiga-
tions.
In summary, this was the first study that demonstrated that VDR
ApaI and BsmI polymorphisms were associated with muscle iso-
kinetic contraction strength in healthy Han Chinese women.
Congruously with previous studies, the evidence supported the
notion that VDR gene polymorphisms were associated with mus-
cular strength. However due to the comparatively small sample
size and because the mechanisms of such influences remain un-
known, further research would be desirable.
Wang P et al. VDR Gene and Muscular Strength In Chinese … Int J Sports Med 2006; 27: 182 – 186
Acknowledgements
The research work was undertaken in Tianjin Research Institute
of Sports Medicine, China.
p
We thank Prof. Shi Zhou and Prof. Yong Zhang for their helpful
assistance in the preparation of the manuscript.
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