缺氧诱导NHE1参与calpain介导ABCA1降解及胆固醇逆转运障碍

项目名称： 缺氧诱导NHE1参与calpain介导ABCA1降解及胆固醇逆转运障碍

项目编号： No.31260250

项目类型： 地区科学基金项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 莫显刚

作者单位： 贵阳医学院

项目金额： 52万元

中文摘要： ABCA1是胆固醇逆转运(RCT)的守门者，具有抗动脉粥样硬化（AS）作用。斑块缺氧广泛存在，伴随RCT功能紊乱，但机制不明。研究证实缺氧诱导NHE1及 calpain活性增加，calpain介导ABCA1降解。故推测缺氧诱导NHE1表达，调节calpain活性，参与ABCA1降解、RCT紊乱及AS形成。为此，首先明确斑块缺氧及NHE1表达，探讨缺氧巨嗜细胞NHE1表达及活性对泡沫细胞形成及胆固醇外流的影响。其次，研究siRNA下调NHE1表达对calpain活性及ABCA1降解影响。最后，从pH、Ca++及Ezrin角度探讨calpain活性改变对ABCA1降解的机制，明确缺氧条件下NHE1、calpain及ABCA1相互作用及细胞内共定位改变。本研究可望阐明缺氧及pH调控通过RCT改变参与AS斑块形成新机制，防治AS及相关疾病提供新思路，开发NHE1相关药物奠定实验基础。

中文关键词： 钠氢交换体-1；ATP结合盒转运蛋白A1；钙蛋白酶；细胞内钙离子；动脉粥样硬化

英文摘要： ABCA1 is thought to be the gatekeeper in reverse cholesterol transport and be beneficial to antiatherosclosis. Severe hypoxia exists in atherosclerotic plaque，accompanying with compromised reverse cholesterol transport, but its mechanisms remain to be elucidated. Calpain mediates ABCA1 degradation.Our previous study demonstrated that hypoxia increased activity of NHE1 and calpain. Thus，we hypothesize that hypoxic induction of NHE1 expression is involved in the ABCA1 degradation, reverse cholesterol transport and atherogenesis. To address the issue, firstly,we examine the relationship of hypoxia and NHE1 expression in atherosclerotic plaques and then the effects of NHE1 expression and activity on foam cell formation and cholesterol efflux. Secondly, the ABCA1 degradation and calpain activity is to explore after knockdown of NHE1 by small inferring RNA. Thirdly, the interaction and subcellular localization of NHE1, calpain and ABCA1 is investigated using coimmunoprecipitation or immunofluorescence and then ABCA1 degradation and calpain activity are detected, after the treatment with NHE1 inhibitor, calcium chelating agent and Ezrin knockdown. This study will be to further insight the novel molecular mechanism whereby hypoxia and pH regulation participate in atherogenesis due to compromised reverse cholesterol tran

英文关键词： Sodium/HydrogenExchanger 1；ATP-binding cassette transporter 1；calpain；intracellular calcium ion；atherosclerosis