固本化瘀通络法调控糖尿病肾病足细胞自噬异常的分子机制研究

项目名称： 固本化瘀通络法调控糖尿病肾病足细胞自噬异常的分子机制研究

项目编号： No.81473522

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： 马跃荣

作者单位： 成都中医药大学

项目金额： 72万元

中文摘要： 业已证实，糖尿病肾病（DN）是一种足细胞疾病，自噬异常是DN足细胞损伤的核心病理环节，但其机制尚未明确。本研究在固本化瘀通络法治疗以脾肾亏虚、痰瘀互结、脉络阻滞为关键病机的DN取得确切疗效的基础上，基于前期对蛋白泛素化降解和内质网应激的研究积累，提出自噬异常是DN足细胞损伤的核心病机，固本化瘀通络法基于整体性调控该环节而起效之假说。以H-2Kb-tsA58 转基因小鼠足细胞株为细胞模型、db/db小鼠为在体模型，以足突蛋白和凋亡为足细胞损伤的标志，以自噬体的形态、数量、活性及相关基因为自噬改变的指标，以mTOR、AMPK、Sirt1、Bcl-2-beclin-1、ERS和氧化应激为自噬信号调控通路,利用激光共聚焦+免疫荧光、双染流式细胞术及生物信息学等研究手段,从整体、细胞及分子水平研究固本化瘀通络法治疗DN的作用靶点、分子药理学机制和自噬信号通路调控模式，拟为DN防治提供新策略。

中文关键词： 糖尿病肾病；足细胞；自噬；固本化瘀通络法

英文摘要： Researches had confirmed that DN(Diabetic Nephropathy) was caused by podocyte damage, and the core pathological procedure was due to be podocyte abnormal autophagy, but the mechanism of which is not clear. We have proved that Guben Huayu Tongluo Principle was effctive to DN causing by insufficient of both the spleen and the kidney and blood stasis and phlegm accumulation. On the basis of preliminary study on protein degradation duing to ubiquitination and endoplasmic reticulum stress(ERS), we assume that podocyte abnormal autophagy is the core pathological process of podocyte indury in DN, and Guben Huayu Tongluo Principle play a part in DN by regulating the overall pathological procedure. To verify the hypothesis, we will use H-2Kb-tsA58 transgenic mice podocye line and db/db mice to test changes on the form, number, activity of autophagosome and related gene, detect related protein for podocyte damage and apoptosis, chose mTOR, AMPK, Sirt1, Bcl-2-beclin-1, ERS and oxidative stress as the targets for autophagy regulation, and utilize several kinds of technique, such as confocal laser, immuno fluorescence, double staining flow cytometry, bioinformatics, etc. From all of above, we can explore the therapy target of Guben Huayu Tongluo Principle in DN, as well as the molecular mechanism and mode of cell signal regulation. This could provide new strategy of preventing and treating DN.

英文关键词： diabitic nephropathy;podocyte;autophagy;gubenhuayutongluo principle