醛固酮对成骨细胞增殖分化及成骨相关基因表达的影响

发布时间：2018-08-06 14:11

【摘要】：目的探讨外源性醛固酮对大鼠原代培养的成骨细胞增殖、碱性磷酸酶活性及成骨相关基因表达的影响及其机制。方法采用酶消化法体外培养大鼠成骨细胞,CCK-8试剂盒和AKP试剂盒分别检测成骨细胞的增殖和碱性磷酸酶活性情况,半定量RT-PCR和Western blotting检测成骨细胞骨形成活性相关标志物和上皮钠离子通道(α-ENaC)基因的mRNA和蛋白表达。结果与对照组相比,醛固酮浓度为1×10~(-2)~1 μmol/L范围内可以促进成骨细胞增殖,浓度为1×10~(-3) μmol/L和10 μmol/L时对成骨细胞增殖无显著影响;但醛固酮在1×10~(-3)~10 μmol/L浓度范围内对成骨细胞碱性磷酸酶活性均无明显影响。与对照组相比,醛固酮浓度为1×10~(-2)~1 μmol/L时均能上调成骨细胞中成骨细胞骨形成活性相关标志物I型胶原蛋白(Coll-Ia)、碱性磷酸酶(ALP)、骨桥蛋白(OPN)和α-ENaC基因的mRNA和蛋白的表达水平(P0.05)。结论醛固酮(1×10~(-2)~1 μmol/L)能明显促进成骨细胞增殖并上调成骨细胞骨形成活性相关标志物的表达,且同时上调ENaC基因的表达,提示ENa C可能是醛固酮调节成骨细胞功能的作用靶点之一。
[Abstract]:Objective to investigate the effect and mechanism of exogenous aldosterone on proliferation, alkaline phosphatase activity and osteoblast related gene expression in primary cultured rat osteoblasts. Methods the proliferation and alkaline phosphatase activity of osteoblasts were detected by CCK-8 kit and AKP kit by enzyme digestion in vitro. Semi-quantitative RT-PCR and Western blotting were used to detect the expression of mRNA and protein in osteoblast bone formation activity and epithelial sodium channel (伪 -ENAC) gene. Results compared with the control group, the concentration of aldosterone in the range of 1 脳 10 ~ (-2) mol/L could promote the proliferation of osteoblasts, but the concentration of 1 脳 10 ~ (-3) 渭 mol/L and 10 渭 mol/L had no significant effect on the proliferation of osteoblasts. However, aldosterone had no significant effect on alkaline phosphatase activity of osteoblasts in the concentration range of 1 脳 10 ~ (-3) ~ (-3) ~ (10) 渭 mol/L. When aldosterone concentration was 1 脳 10 ~ (-2) or 1 渭 mol/L, the expression levels of type I collagen (Coll-Ia), (ALP), osteopontin (OPN) and 伪 -ENAC gene mRNA and protein in osteoblasts were up-regulated (P0.05). Conclusion Aldosterone (1 脳 10 ~ (-2) ~ (-1) 渭 mol/L) can significantly promote the proliferation of osteoblasts and up-regulate the expression of markers related to osteogenesis activity of osteoblasts, and also up-regulate the expression of ENaC gene, suggesting that ENa C may be one of the targets for aldosterone to regulate the function of osteoblasts.
【作者单位】：广东药科大学生命科学与生物制药学院;广东药科大学生物资源与创新药物研究中心;
【基金】：国家自然科学基金青年基金项目(81400853) 广东省普通高校国际暨港澳台合作创新平台及国际合作重大项目(项目编号2015KGJHZ022) 中央财政支持地方高校发展专项资金(粤财政[2016]202号)~~
【分类号】：R580

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