mad2b在高糖诱导肾小球内皮细胞损伤中作用word论文.docVIP

优秀毕业论文 精品参考文献资料 英文缩略词表 英文缩写 英文全称 DN diabetic nephropathy GEnC glomerular endothelial cell MAD2B mitotic arrest deficient 2-like protein2 ESRD End stage real disease DMEM Dulbeccos Modified Eagles medium Ctrl control HG high glucose scra scramble APC/C anaphase-promoting complex/cyclosome BCA bicinchonininc acid DMSO dimethyl sulfoxide SDS sodium dodecyl sulfate PAGE polyacrylamide gel electrophoresis eNOS endothelial nitric oxide synthase NO nitric oxide AngII angiotensin II TGF-β Transforming Growth Factor IL-6 interleukin- 6 Cdh1 E-cadherin Cdc20 cell division cycle 20 shRNA short hairpin Ribonucleic Acid TCF4 Transcription factor 4  MAD2B 在高糖诱导的肾小球内皮细胞损伤中的作用 华中科技大学同济医学院附属协和医院肾内科 硕士研究生 郝雨 导 师 张春 中文摘要 目的：探讨 MAD2B 在高糖诱导的肾小球内皮细胞损伤中的作用。 方法：体外培养小鼠肾小球内皮细胞，Western Blot 检测高糖（30mM）刺激下不同时 间点 MAD2B 蛋白的表达；脂质体法将重组质粒 MAD2B shRNA 和 scrambled shRNA 转染 GEnC，实验分为对照组、高糖组、高糖+scramble 组、高糖+MAD2B shRNA 组， 检测各组细胞的凋亡率、增殖活力、NO 生成水平以及单层细胞的通透性。 结果：高糖刺激肾小球内皮细胞 MAD2B 的表达量明显升高；脂质体法转染 MAD2B shRNA，显著抑制 MAD2B 的表达；和对照组相比，高糖组凋亡率升高，而 MAD2B shRNA 干预组凋亡减轻；高糖刺激 48h 细胞增殖活力减弱，但高糖+shRNA 组与高糖 +scramble 组相比细胞活力较高；高糖刺激 24h、48h 细胞生成 NO 水平较对照组降低， 但高糖+shRNA 组较高糖+scramble 组 NO 分泌较多；高糖组单层细胞通透性升高， MAD2B shRNA 干预组能减弱高糖环境下内皮的高通透性反应。 结论：高糖诱导内皮细胞 MAD2B 表达升高，抑制 MAD2B 的表达可以减轻高糖条件 下内皮细胞的凋亡和高通透性，增加细胞增殖活力和 NO 生成。 关键词：MAD2B；高糖；肾小球内皮细胞  The role of MAD2B in high glucose induced glomerular endothelial cells injury Post graduate：Hao Yu Tutor：Prof. Zhang Chun Department of Nephrology ,Union Hospital, Tongji Medical College Huazhong University of Science and Technology Abstract Objective: To explore the role of MAD2B in high glucose-induced glomerular endothelial cells injury. Methods: Mouse glomerular endothelial cells (GEnC) were incubated in normal glucose (5.5 mmol/l) or high glucose (30mmol/l). GEnC was transfected separately with MAD2B shRNA and scrambled shRNA by Lipofectamine 2000.according to manufacture’s protocols under high glucose. Cells were divided into controlled group, high glucose group, high glucose +sc