c-FLIP调控细胞坏死性凋亡在糖尿病肾病进展中的作用及机制研究

项目名称： c-FLIP调控细胞坏死性凋亡在糖尿病肾病进展中的作用及机制研究

项目编号： No.81460146

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 颜晓勇

作者单位： 遵义医科大学

项目金额： 47万元

中文摘要： 糖尿病肾病(Diabetic nephropathy,DN)早期就伴随有足细胞损伤，进而可导致蛋白尿及肾小球硬化。坏死性凋亡是近年发现的一种新型细胞程序化死亡方式，可能导致DN的足细胞和肾组织损伤。研究发现c-FLIP在肾皮质和肾小管上皮细胞均有表达，并且在调控坏死性凋亡途径中起着至关重要的作用。坏死性凋亡在DN进展中的作用以及受c-FLIP调控的机制需要深入研究。高糖刺激大鼠足细胞以及建立1型DN模型，采用电镜、real-time PCR、免疫组化、激光共聚焦及western-blot检测足细胞形态和RIP1、RIP3、c-FLIP、podocin、nephrin、IL-6及NF-kB的核酸和蛋白表达。敲除或调高c-FLIP的表达后，同样检测以上指标。了解坏死性凋亡是否介导了DN大鼠的足细胞和肾组织损伤；增加c-FLIP的表达是否通过减轻细胞坏死性凋亡起到保护DN肾脏的作用。

中文关键词： 糖尿病肾病；足细胞；坏死性凋亡；c-FLIP蛋白；发病机制

英文摘要： Diabetic nephropathy(DN) is early accompanied by podocyte injury that can result in proteinuria and glomerulosclerosis.A new mode of programmed cell death has recently been described, termed necroptosis.It may cause the injury, death and detach of podocyte and the tissue damage in DN.c-FLIP expression was found in kidney cortex and tubular epithelial cells,c-FLIP play an essential role in the regulation of necroptosis.The mechanism of necroptosis in DN is still unclear. Further research is required to determine the specific role of necroptosis and the regulation by c-FLIP.The rat podocytes were treated by high glucose and Sprague- -Dalwley(SD) rats by streptozotocin.The expression of RIP1、RIP3、c-FLIP、podocin、nephrin、IL-6 and NF-kB in podocytes and renal tissue were detected by real-time PCR、immunohisochemistry、laser confocal microscopy and western blot analysis; Electron microscopy was used to explore the cellular morphology of podocytes.We tested the same indicators when the podocytes treated with Nec-1 and diabetic Rats with anti IL-2R antibody.The aim of this study was to confirm the effects of the necroptosis pathway activation on renal cell injury to DN and whether blocking necroptosis pathway could attenuate this injury through increasing expression of c-FLIP in vitro and in vivo.

英文关键词： Diabetic nephropathy;podocyte;necroptosis;c-FLIP;mechanism