TRPV5介导雌激素抑制破骨细胞骨吸收作用及机制研究

项目名称： TRPV5介导雌激素抑制破骨细胞骨吸收作用及机制研究

项目编号： No.81472129

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 叶添文

作者单位： 中国人民解放军第二军医大学

项目金额： 72万元

中文摘要： 雌激素（E2）具有抑制破骨细胞骨吸收作用，但其确切机制仍不清楚。我们前期发现：E2通过上调破骨细胞TRPV5的表达，抑制其骨吸收能力。雌激素受体(ER)阻滞剂可抑制E2对破骨细胞TRPV5表达的刺激作用。TRPV5基因启动子区域无经典ERE信号通路必需的雌激素反应元件，而含有AP-1、SP1和NF-κB等非经典ERE信号通路转录调控因子结合位点。因此，推测E2通过非经典ERE信号通路调控破骨细胞TRPV5表达。本课题拟以破骨细胞TRPV5表达调控为线索，通过RNAi、ChIP、生物信息学技术等手段，首先，探讨并确定TRPV5介导E2抑制破骨细胞骨吸收作用；其次，阐明E2通过ER调控破骨细胞TRPV5表达；最后，明确雌激素受体非经典ERE信号通路在E2调控破骨细胞TRPV5基因转录激活中的作用。籍此阐释TRPV5介导E2抑制破骨细胞骨吸收作用及分子机制，进一步揭示E2防治骨质疏松的理论依据。

中文关键词： 破骨细胞；瞬时性受体电位通道香草酸受体亚型5；雌激素；骨吸收；信号通路

英文摘要： The precise molecular events underlying the inhibitory effect of estrogen (E2) on osteoclastic bone resorption remained elusive. Our previous studies showed that E2 inhibited osteoclastic resorption by increasing the expression of TRPV5 channel in osteoclasts. The estrogen receptor (ER) blocker significantly attenuated the stimulatory effect of estrogen on TRPV5 expression. The promoter sequence of the TRPV5 gene does not consist of estrogen response elements (ERE) which are required in the classical ERE genomic signaling pathway. However, the TRPV5 gene promoter contains several of these AP-1, SP1 and NF-κB sites, which can mediate the transcriptional activation of estrogen-liganded ER through non-classical genomic pathway. On the basis of this information, we presume that E2 regulates the expression of TRPV5 in the osteoclasts through the non-classical genomic signaling pathway. Therefore, our research is centralized with the regulation of the expression of TRPV5 in the osteoclasts. RNA interference(RNAi), chromatin immunoprecipitation(ChIP) and bioinformatics are applied. Firstly, TRPV5-mediated the inhibitory effect of E2 on the osteoclastic bone resorption will be confirmed. Sencondly, the regulation of E2 on the expression of TRPV5 in the osteoclasts by estrogen receptor (ER) will be clarificated. Finally, the regulatory effect of E2 on the TRPV5 gene activation and transcription is mediated through the ER dependent non-classical genomic pathway will be revealed. According to this study, the molecular mechanisms that TRPV5 mediates the inhibitory effect of E2 on the osteoclastic bone resorption will be found. More importantly, the application of E2 to prevent and treat osteoporosis will gain more scientific evidences from this study.

英文关键词： Osteoclasts;TRPV5;estrogen;bone resorption;signaling pathway