芳香化酶和雌激素相关受体在人骨髓间充质干细胞中的表达  

Expression of aromatase and estrogen-related receptors in human bone marrow mesenchymal stem cells

作　　者：魏秋实[1,2] 陈镇秋[1,3] 何伟[1,3] 邓伟民[2] 王海彬[1,3] 黄世金[3] 郭承[3] 

机构地区：[1]广州中医药大学第一附属医院骨科,广东省广州市510407 [2]解放军广州军区广州总医院康复科博士后科研工作站,广东省广州市510010 [3]广州中医药大学国家重点学科中医骨伤科实验室,广东省广州市510405

出　　处：《中国组织工程研究》2015年第36期5758-5763,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金青年科学基金项目(81302994)<补肾药通过TAZ与经典Wnt通路crosstalk调控肾虚型骨髓基质干细胞成骨能力的机制>;国家自然科学基金面上项目(81473697)<不同证型股骨头坏死与MRI;病理及分子分型相互关系的探讨研究>;广东省自然科学基金博士启动项目(S2013040014927)<补肾药靶向TAZ/β-catenin调控肾虚型BMSCs成骨分化的实验研究>~~

摘　　要：背景:骨髓间充质干细胞中可能存在芳香化酶和雌激素受体相互作用的雌激素信号途径,调控骨髓间充质干细胞生物活性。目的:观察成人骨髓间充质干细胞成骨诱导分化过程中芳香化酶和雌激素相关受体的表达。方法:将骨髓间充质干细胞分为单纯培养组和成骨诱导组,分别用低糖DMEM完全培养基和成骨诱导分化完全培养基培养,MTT检测细胞增殖能力,茜素红染色观察成骨矿化能力,q PCR和Western blot检测芳香化酶、雌激素受体α、雌激素受体β和雌激素受体相关受体αm RNA和蛋白表达,ELISA检测细胞上清液和裂解液中雌二醇水平。结果与结论:培养72 h,成骨诱导组增殖能力最强;培养21 d,成骨诱导组茜素红染色显示大量钙沉积;q PCR和Western blot结果显示成骨诱导组能促进芳香化酶和雌激素受体αm RNA和蛋白表达,抑制雌激素受体相关受体αm RNA和蛋白表达,成骨诱导组雌激素受体βm RNA和蛋白表达与单纯培养组无差异;ELISA结果显示成骨诱导组上清液中雌二醇水平高于裂解液及单纯培养组上清液中雌二醇水平。结果表明成人骨髓间充质干细胞及成骨诱导分化后能表达芳香化酶、雌激素受体α、雌激素受体β和雌激素受体相关受体α,亦能自身合成和分泌雌激素,由此产生的雌激素可能是通过相关受体对骨髓间充质干细胞成骨分化发挥作用。BACKGROUND： Estrogen signaling pathway for interaction between aromatase and estrogen-related receptor may exist in bone marrow mesenchymal stem cells, which is used for regulating biological activity of bone marrow mesenchymal stem cells. OBJECTIVE： To observe the expression of aromatase and estrogen-related receptors in adult bone marrow mesenchymal stem cells during osteogenic differentiation. METHODS： Bone marrow mesenchymal stem cells were respectively cultured in low-glucose DMEM medium（control group） and osteogenic induction medium（induction group）. Cell proliferation and calcium deposition were determined by MTT assay and alizarin red staining, respectively. The expression of aromatase, estrogen receptor α, estrogen receptor β, and estrogen-related receptor α during osteogenic differentiation were determined by real-time PCR and western blot analysis. Estradiol levels in supernatants and lysates were detected by ELISA method. RESULTS AND CONCLUSION： In the induction group, the proliferation ability of bone marrow mesenchymal stem cells was the strongest at 72 hours of culture; while there were a great amount of calcium nodules formed at 21 days of culture. Results from PCR and western blot assay showed that the expression of aromatase and estrogen receptor α was improved in the induction group, but the expression of estrogen-related receptor α was inhibited. There was no difference in the expression of estrogen receptor β between the two groups. ELISA results indicated that the level of estradiol in the supernatant of induction group was the highest. These findings indicate that aromatase, estrogen receptor α, estrogen receptor β and estrogen-related receptor α are all involved in osteogenesis of bone marrow mesenchymal stem cells. Moreover, estradiol can be synthesized and secreted in bone marrow mesenchymal stem cells, and most likely, promote the osteogenic differentiation of bone marrow mesenchymal stem cells by related receptor pathway.

关 键 词：干细胞 骨髓干细胞 芳香化酶 骨髓间充质干细胞 成骨诱导分化 雌激素受体 雌激素相关受体 骨代谢疾病 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]