AMPK-Beclin-1/Vps34通路在维生素D3（Vit D)诱导足细胞自噬中的作用和机制研究

项目名称： AMPK-Beclin-1/Vps34通路在维生素D3（Vit D)诱导足细胞自噬中的作用和机制研究

项目编号： No.81470950

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： 肖厚勤

作者单位： 湖北医药学院

项目金额： 66万元

中文摘要： 自噬和凋亡是足细胞最重要的两个病理生理过程。高水平的自嗜对维持足细胞稳态具有重要意义。Beclin-1/Vps34是自噬溶酶体形成关键，Bcl-2与Beclin-1结合阻碍Beclin-1/Vps34复合物形成而抑制自噬。前期研究证实VitD可以通过抑制TRPC6和RNAK的表达抑制凋亡，而凋亡信号和自噬信号存在串话作用,自噬抑制可促进凋亡。VitD对凋亡的抑制是否是通过活化自噬实现的呢？本课题以足细胞损伤的细胞、动物模型，通过confocal观察GFP-LC3转染足细胞在应激状态下自噬的变化；通过游离钙荧光探针、膜片钳技术观察VitD对细胞Ca2+通道和Ca2+浓度的影响；通过基因转染、基因沉默观察VitD对Beclin-1、Vps34、AMPK表达的影响，来探讨AMPK-Beclin-1/Vps34通路在VitD诱导足细胞自噬中的作用，为阻止足细胞损伤，寻找激活自噬的药物提供理论。

中文关键词： 维生素D3；足细胞损伤；自噬；Beclin-1；凋亡

英文摘要： Autophagy and apoptosis are the two most important pathophysiological processes of podocytes and high level of autophagy plays an key role to maintain podocyte homeostasis. Recent researches have showed the Beclin-1/Vps34 complex was a key factor in regulation of the formation of autolysosome and the combination of BCL-2 and Beclin-1 inhibit Autophagy due to defect of Beclin-1/Vps34 complex formation. Previous studies show VitD could inhibit the apoptosis of podocytes through suppressing the expression of TRPC6 and RNAK. There exists crosstalk between autophagy and apoptosis, the activation of autophagy could inhibit apoptosis. Thus, could VitD inhibit the apoptosis of podocytes through the activation of autophagy? To confirm this propose, the autophagy process of podocytes under the stress was observed by confocal through the transfection of GFP-LC3 under the level of cell and animal, and the change of the Ca2+ Channel and Ca2+ concentration of podocytes induced by VitD were detected by Ca2+ -mark- fluorescent probes and patch clamp technique. To investigate the role of AMPK-Beclin-1/Vps34 passway in podocyte autophagy and apoptosis, the expression level of Beclin-1，Vps34 and AMPK in VitD induced podocytes were detected by gene transfection and silence. The study of this project will allow us to gain insights into the effect and mechanism of AMPK- Beclin-1/Vps34 pathway in the Vitamin D3 Induced podocyte Autophagy and provide new theoretical basis for finding drugs to promote autophagy and podocyte protection.

英文关键词： vitamin D;podocyte injury;autophagy;Beclin-1;apoptosis