α玉米赤霉醇对小鼠骨髓间充质干细胞向成骨细胞分化过程中OPG和RANKL蛋白表达的影响  

The effect of a-zearalanol on the expression of OPG and RANKL during the osteoblast differentiation by bone marrow derived mesenchymal stem cells in mice

作　　者：方晔[1] 宗少晖[1] 赵劲民[2] 曾高峰[3] 邹斌[4] 彭金珍 

机构地区：[1]广西医科大学第一附属医院脊柱骨病外科,广西南宁530021 [2]广西医科大学第一附属医院骨科,广西南宁530021 [3]广西医科大学公共卫生学院,广西南宁530021 [4]广西医科大学研究生学院,广西南宁530021

出　　处：《中国骨质疏松杂志》2015年第2期163-167,174,共6页Chinese Journal of Osteoporosis

基　　金：广西自然科学基金资助项目(2014GXNSFAA118400)

摘　　要：目的探讨α-玉米赤霉醇(α-zearalanol,α-ZAL)对小鼠骨髓间充质干细胞(Bone marrow-derived mesenchymal stem cells,BMSCs)向成骨细胞增殖、分化的影响。方法采用全骨髓培养差速贴壁法体外分离培养小鼠骨髓间充质干细胞,取第3代细胞分为空白对照组、雌二醇阳性对照组、低浓度α-ZAL组、中浓度α-ZAL组,高浓度α-ZAL组。镜下每天观察细胞形态变化,碱性磷酸酶染色鉴定成骨细胞纯度,MTT实验测定细胞增殖活性绘制细胞生长曲线,采用酶联免疫吸附法(ELISΑ)测定碱性磷酸酶(alkaline phosphatase,ALP)、骨形态发生蛋白2(Bone morphogenetic protein-2,BMP-2),运用蛋白免疫印迹法(Western blotting,WB)检测护钙素(Osteoprotegerin,OPG)和NF-KB受体活化配体(Receptor actiator of nuclear factor-KB ligand,RANKL)的蛋白水平表达变化。结果不同浓度的α-玉米赤霉醇可显著的促进小鼠骨髓间充质干细胞向成骨细胞增殖、分化,增加细胞活性(P<0.05),明显提高小鼠骨髓间充质干细胞向成骨细胞分化后ALP和BMP-2的表达(P<0.05),并且显著的上调了成骨细胞内OPG/RANKL的表达率(P<0.05)。结论不同浓度的α-玉米赤霉醇可以促进骨髓间充质干细胞向成骨细胞增殖、分化,并且通过上调OPG/RANKL的表达率抑制成骨细胞细胞向破骨细胞分化,有望成为临床骨折疏松症治疗的激素替代药物。Objective To observe the effect of a-zearalanol on the osteogenic differentiation by mouse bone marrow derived mesenchymal stem cells（BMSCs）.Methods Mouse BMSCs were isolated using whole bone marrow adherence method.After the third generation,the BMSCs were randomly placed into the following subgroups：Control group,OM group,17β-E2 group,a—ZAL 10^（-7） mol/L group,α-ZAL 10^（-6） mol/L group,and α-ZAL 10^（-5） mol/L group.An inverted microscope was employed to observe the morphological changes of the cells.ALP staining was performed to identify the purity of osteoblasts.MTT test was used to determine the cell proliferation and to draw the growth curve.ALP and BMP-2 were examined using ELISA method.Western blotting was used to determine the expression of osteoprotegrin（OPG） and RANKL.Results The proliferation and differentiations of osteoblasts by BMSCs were stimulated by different concentrations of a-zearalanol（P〈0.05）.ALP activity and BMP-2 were significantly increased（P〈0.05）.The expression of OPG/RANKL was also up-regulated in osteoblasts（P〈0.05）.Conclusion α-ZAL can obviously increase the proliferation and differentiations of osteoblasts by BMSCs and can effectively suppress osteoclastogenesis formation by un-regulation of OPG/RANKL.Therefore,it would be an alternative of hormones in the treatment of osteoporosis.

关 键 词：α玉米赤霉醇 骨髓间充质干细胞 成骨细胞 骨形态发生蛋白2护钙素 

分 类 号：R651.2[医药卫生—外科学]