鲑鱼降钙素对去卵巢大鼠骨髓细胞OPG、RANKL基因和蛋白表达的影响

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[目的]观察鲑鱼降钙素(sCT)对去卵巢大鼠骨密度(BMD)、血清I型胶原交联羧基末端肽(ICTP)变化的影响,以及骨髓细胞骨保护素(OPG)和核因子κB受体活化因子配体(RANKL)的基因表达和两者在胫骨骨骺端蛋白含量的变化。[方法]取3个月龄雌性SD大鼠24只,随机平均分3组:假手术组(Sham)、鲑鱼降钙素处理组(sCT)、安慰剂组(OVX)。采用双侧卵巢切除术复制骨质疏松大鼠模型。术后2周CT组予鲑鱼降钙素皮下注射12周,应用双能X线吸收仪法(DXA)测BMD,ELISA法测量血清ICTP浓度,qRT-PCR法定量骨髓细胞OPG和RANKL的mRNA表达量,免疫组织化学染色法测定胫骨干骺端OPG和RANKL蛋白表达量。[结果]与OVX组比较,sCT组的腰椎BMD上升显著(P<0.05),但股骨BMD改变不明显(P>0.05);血清ICTP含量显著降低(P<0.05);骨髓细胞RANKL的mRNA表达量变化不大(P>0.05),但OPG的mRNA表达量升高(P<0.05),OPG/RANKL的比率升高(P<0.05);胫骨干骺端也呈现出RANKL蛋白改变不明显(P>0.05),而OPG的蛋白分泌增加(P<0.05),从而OPG/RANKL的比率高于OVX组(P<0.05)的现象。[结论]降钙素可以预防腰椎BMD的丢失,降低血清ICTP水平,在体内可能主要通过上调OPG的mRNA表达和蛋白分泌,影响OPG/RANKL/RANK系统,影响破骨细胞功能,抑制骨吸收,进而达到预防绝经后骨质疏松的目的。 [Objective] To observe the effect of salmon calcitonin (sCT) on the changes of bone mineral density (BMD) and the type Ⅰ collagen cross-linked carboxyl terminal peptide (ICTP) in ovariectomized rats and the effects of osteoprotegerin (OPG) and nuclear factor κB receptor activator ligand (RANKL) gene expression and the content of both proteins in the tibial epiphysis. [Method] Totally 24 female SD rats of 3 months old were randomly divided into 3 groups: sham operation group, salmon calcitonin treatment group (sCT) and placebo group (OVX). Bilateral ovariectomized rat models of osteoporosis were replicated. At 2 weeks after operation, the patients in CT group were given salmon calcitonin subcutaneously for 12 weeks. BMD was measured by dual-energy X-ray absorptiometry (DXA), the serum ICTP concentration was measured by ELISA. The mRNA expression of OPG and RANKL in bone marrow was measured by qRT-PCR Quantities of OPG and RANKL protein in metaphyseal tibia were detected by immunohistochemical staining. [Results] Compared with OVX group, BMD of lumbar spine of sCT group increased significantly (P <0.05), but BMD of femur did not change significantly (P> 0.05), and serum ICTP level decreased significantly (P <0.05) (P> 0.05), but the mRNA level of OPG increased (P <0.05) and the ratio of OPG / RANKL increased (P <0.05). The change of RANKL protein in metaphyseal tibia was not significant P> 0.05), while the secretion of OPG increased (P <0.05), and the ratio of OPG / RANKL was higher than that of OVX group (P <0.05). [Conclusion] Calcitonin can prevent the loss of BMD in lumbar spine and decrease the level of ICTP in serum. It may affect OPG / RANKL / RANK system mainly by up-regulating mRNA and protein secretion of OPG, affecting osteoclast function, inhibiting bone resorption, Then achieve the purpose of preventing postmenopausal osteoporosis.
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