mTOR信号通路在腹膜纤维化中的作用研究

Peritoneal fibrosis inducing ultrafiltration failure is one of the important reasons for technique failure in PD patients, so inhibiting peritoneal fibrosis will significantly improve the long-term prognosis of PD patients. Several experiments have confirmed that the mTOR signaling pathway and rapamycin have a clear relationship with fibrosis in a variety of tissues and organs. Moreover, our group had clearly certificated that rapamycin can significantly reduce peritoneal fibrosis. In order to further determine the mechanisms and the role of mTOR signaling pathway in peritoneal fibrosis, This study will verify the expression of mTOR signaling pathway in vivo and vitro model. Rapamycin will be used to intervene the vivo and vitor model of peritoneal fibrosis to probe the relationship of p-P70S6K's decrease with peritoneal fibrosis and EMT. Plasmid and siRNA will be used to regulate the expression of mTOR to further determine that the mTOR signaling pathway plays an important role in peritoneal fibrosis. The purpose of the research is to provide a new direction and research targets for the prevention and treatment of peritoneal fibrosis.

腹膜纤维化导致超滤衰竭是PD患者技术失败的重要原因之一，因此抑制腹膜纤维化将显著改善PD患者的长期预后。多项实验已证实mTOR信号通路、雷帕霉素与多种组织器官的纤维化明确相关，而且本课题组已明确证明雷帕霉素能显著减轻腹膜纤维化。为进一步确定mTOR信号通路和雷帕霉素在腹膜纤维化中的机制和作用。本研究将在体内、外腹膜纤维化模型中验证mTOR信号通路的表达情况。应用雷帕霉素干预体内、外腹膜纤维化模型探讨雷帕霉素下调p-P70S6K的表达与腹膜纤维化和EMT的关系。应用质粒或siRNA调节mTOR的表达，进一步确定mTOR信号通路在腹膜纤维化起到重要的作用。为腹膜纤维化的防治提供新的方向和研究靶点。

腹膜纤维化导致超滤衰竭是PD患者技术失败的重要原因之一，因此抑制腹膜纤维化将显著改善PD患者的长期预后。多项实验已证实mTOR信号通路、雷帕霉素与多种组织器官的纤维化明确相关，而且本课题组已明确证明雷帕霉素能显著减轻腹膜纤维化。为进一步确定mTOR信号通路和mTOR阻断剂在腹膜纤维化中的机制和作用。本研究在体内、外腹膜纤维化模型中验证mTOR信号通路的表达情况。应用雷帕霉素和BEZ235 干预体内、外腹膜纤维化模型探讨mTOR阻断剂下调p-P70S6K表达与腹膜纤维化、EMT的关系。同时，在国内、外首次对腹膜纤维化模型进行蛋白质谱iTRAQ分析，为寻找新的干预治疗靶点提供方向。研究结果显示：1）体外实验：在 TGF-β刺激下，人腹膜间皮细胞发生纤维化改变、mTOR信号通路被激活。mTOR阻断剂雷帕霉素能够显著抑制p-P70S6K表达，并不引起p-AKT负反馈上调。雷帕霉素可以显著抑制TGF-β所致的纤维化改变。2）体内实验： 采用mTOR阻断剂治疗腹膜纤维化大鼠，结果发现mTOR阻断剂（雷帕霉素、BEZ235）可以显著抑制腹膜纤维化和mTOR信号通路的活化。蛋白质谱iTRAQ分析显示COL,Fn表达在腹膜纤维化模型显著增加；予以mTOR阻断剂干预后COL,Fn表达显著下调。各类差异蛋白正在进一步整理分析，以探索新的干预治疗靶点。本研究证实mTOR信号通路在腹膜纤维化中起到重要作用，mTOR阻断剂能够抑制腹膜纤维化的发生、发展。通过iTRAQ检测希望能够找到防治腹膜纤维化新的方向和治疗靶点。