ERβ通过调控OPG/RANK/RANKL信号通路调控小鼠成骨细胞的成骨能力

ERβ regulates osteogenic potential of osteoblast through regulation of OPG/RANK/RANKL signaling in mice

目的在细胞水平上,用RNA干扰(RNAi)技术研究雌激素受体β(ERβ)对OPG/RANK/RANKL信号通路的调控,及其调控小鼠成骨细胞的成骨能力。方法取的小鼠成骨细胞株MC3T3-E1为对象,设立3组,其中2个组分别转染重组ERβRNAi载体和阴性对照ERLRNAi载体,第3组为空白对照组。3组在相同条件下培养。然后,检测ERβ基因沉默后小鼠成骨细胞中骨保护素(OPG)、核因子κB配体的受体激活子(RANKL)表达的变化、各组成骨细胞碱性磷酸酶(ALP)活性和各组成骨细胞形成钙结节的差异。用SPSS16.0软件统计分析。方法 RNAi组OPG的Ct值较空白对照组和阴性干扰组小,基因表达增强;RNAi组RANKL的Ct值较其它两组大,基因表达降低;RNAi组的ALP活性较其它两组高。均有显著性差异,P<0.05。RNAi组成骨结节较空白对照组和阴性干扰组的数量较多,体积较大。结论根据ERβ沉默后的结果,可以反向推断,雌激素与ERβ结合,增强ERβ表达,降低OPG的合成和表达,增加RANKL的合成和表达,减少ALP的分泌,进而抑制新骨的生成。

[Objective] At the cellular level, to study whether the ERβregulates the secretion and expression of the cytokine OPG and RANKL and the osteogenic capability of mice osteoblasts. [Methods] The mice osteoblastic cell lines MC3T3-E1 is divided into 3 groups, tow of which groups were transfeeted with recombinant plasmid ERβ RNAi and negative control plasmid ERLRNAi, group 3 was the control group, the same amount of PBS instead of plasmids. 3 groups of cells cultured under the same conditions. In each group, OPG and RANKL expression, alkaline phosphatase （ALP） activity and the calcium nodules were detected. SPSS 16.0 software was used. [ Results ] The results indicate that the OPG Ct value was relatively lower, gene expression was more enhanced. Meanwhile, the Ct value of the RANKL is higher in RNAi group than in other two groups, gene expression was more reduced. ALP activity in the RNAi group was higher than those in other groups. The calcium nodules formed in RNAi group were a larger number and larger volume compared with the control group or negative control group. [ Conclusion ] According to the results of silencing, we can reversely infer that ERβ expression may inhibit the proliferation, differentiation and maturation of osteoblasts, with reducing the secretion of ALP, the expression of OPG and increasing the expres- sion of RANKL, thereby inhibiting the formation of new bone.