Comparative study on the characteristics of bone metabolism in forelimb malformation WHBE rabbits via the OPG/RANK/RANKL system
Received:May 30, 2016  
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DOI:10.3969/j.issn.1005-4847.2016.05.012
KeyWord:WHBE rabbit;Japanese rabbit;Forelimb malformation;Bone metabolism;OPG/RANK/RANKL system
        
AuthorInstitution
吕建敏 浙江中医药大学动物实验研究中心, 杭州
陈方明 浙江中医药大学动物实验研究中心, 杭州
陈诚 浙江中医药大学动物实验研究中心, 杭州
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Abstract:
      Objective To comparatively investigate the characteristics of bone metabolism in forelimb malformation WHBE rabbits based on the OPG/RANK/RANKL system. Methods Healthy male 2-2.5-month old WHBE rabbit (HWR), Healthy Japanese rabbit (HJR) and forelimb malformation WHBE rabbit (FMWR) were used in this study and divided into 3 groups, HWR, HJR and FMWR groups, with 10 rabbits in each group. The shape of forelimb and mean gray value observed from X-ray film were examined, and the bone tissue micro-morphology was analyzed using HE staining. Real-time fluorescence quantitative PCR was used to determine the expression of osteoprotegerin (OPG) and receptor activator of NF-KB ligand (RANKL) mRNA. The expressions of OPG, receptor activator of NF-KB (RANK) and RANKL protein in serum and bone tissue were assayed by enzyme-linked immunoassay (ELISA) and immunohistochemistry, respectively. Results Compared with the HWR and HJR groups, rabbits in the FMWR group appeared abnormal in toutuous forelimb and thinner bone cortex. The mean gray values of X-ray in the FMWR group were lower than that in the HWR group (P<0.05). There were significant differences between the FMWR and healthy rabbits (HWR and HJR) in the following parameters: RANKL mRNA expression and RANKL/OPG mRNA ratio in the liver(P<0.05, P<0.01), serum protein expression of RANK and RANKL and RANKL/OPG ratio (P<0.05, P<0.01), and positive index of expression of RANKL protein and RANKL/OPG ratio in the bone tissue(P<0.05, P<0.01). Furthermore, the gene expression levels of OPG and RANKL of HWR were significantly higher than that of HJR(P<0.05, P<0.01). Conclusions The FMWR show some abnormal symptoms in bone metabolism as well as decrease of bone quality and histological changes of bone micro-structure, due to the significant increase of RANKL/OPG ratio of FMWR. The breed differences between WHBE and Japanese rabbits in RANKL mRNA expression level may be one of factors inducing limb malformation in the WHBE rabbits.
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