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have an effect on femur BMC in OVX rats (Choi and DiMarco
2009
) . Although
it is expected that taurine and arginine act synergistically on bone, the effect of
taurine and arginine simultaneously on bone has not been studied. The objective
of the present study was to investigate the effect of taurine and arginine supple-
mentation with measures of spine and femur BMD and bone markers in growing
female rats.
31.2
Methods
31.2.1
Materials
Forty-eight 6-week-old, female Sprague-Dawley rats were purchased from Bio
Genomics, Oriental, Seoul, Korea. On arrival at our lab, rats were acclimated for 5
days to a standard laboratory nonpurified diet (Samyang, Seoul, Korea). After accli-
mation, the rats were divided into four groups through the use of a randomized
complete block design, with blocks determined by initial body weight, as follows: a
casein-based diet (control), a casein-based diet with an arginine (Arg), a casein-
based diet with a taurine (Tau), and a casein-based diet with an arginine and taurine
(Arg + Tau). Rats were individually housed in stainless steel cages in a room with
controlled temperature (23°C) and humidity (55%) and were given free access to
the experimental diets and water. Rats were maintained at 12-h light (07:00-19:00 h)
and dark cycle. The compositions of the experimental diets are shown in Table
31.1
.
For 9 weeks, rats were fed experimental powdered diets. The experimental diet
groups were fed similar diets which were supplemented with arginine or/and tau-
rine. Blood samples were collected from the abdominal aorta and serums were sepa-
rated at 3,000 rpm for 20 min. Serums were stored at −70°C until analysis. Serum
concentrations of alkaline phosphatase (ALP) and osteocalcin were measured.
Serum calcium and phosphate were also measured. Serum ALP and osteocalcin and
urinary DPD cross-link value were measured as markers of bone formation and
resorption. ALP activity was reported as units per liter (U/L). The concentration of
urine deoxypyridinoline (DPD) was measured with an enzyme immunoassay that
preferentially recognizes the free form of DPD (CLIA, Pyrilinks-D DPC, USA).
DPD was corrected for creatinine excretion (DPD/Cr). The concentration of serum
osteocalcin was measured with an osteocalcin kit (IRMA, OSTEO-RIACT, Cis Bio,
Saclay, France), which recognizes the intact form of osteocalcin. Serum ALP activ-
ity was measured with a kit from Enzymatic assay (Prueauto S ALP) following the
manufacturer's instructions.
BMD and BMC were measured using PIXImus (GE Lunar Co, Wisconsin, USA)
in spine and femur on 9 weeks after feeding. The experimental protocol was
approved by Institutional Animal Care and Use Committee (IACUC) in Keimyung
University and conformed to the Guide for the Care and Use of Laboratory
Animals.
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