内质网应激相关因子Hrd1在糖尿病鼠视网膜的表达变化

发布时间：2018-10-29 22:39

【摘要】：目的:内质网应激在糖尿病视网膜病的发生和发展中起到重要作用,而羟甲基戊二酰辅酶A还原酶降解蛋白1(Hrd1)能通过内质网相关蛋白降解途径稳定内质网内环境从而减少内质网应激的发生。本实验通过基因芯片分析内质网相关因子的变化,并观察Hrd1在糖尿病鼠视网膜和体外高糖培养人视网膜微血管内皮细胞的表达变化,分析它在糖尿病视网膜病变发生和发展中的作用。 方法腹腔注射链脲佐菌素(STZ)(65 mg/kg)制作糖尿病SD大鼠模型,对照组大鼠腹腔注射同等剂量柠檬酸钠溶液。于注射后72小时用鼠尾采血法测量血糖,7天后以血糖值≥16.7mmol/L认定为Ⅰ型糖尿病鼠建模成立,以后每周监测血糖一次。模型成立三个月时取大鼠视网膜组织。用自行设计实时定量PCR芯片分析89个内质网相关因子的表达变化。用Western blot、免疫荧光检测Hrd1在三月糖尿病模型鼠和对照组大鼠视网膜中的表达。对体外培养人视网膜微血管内皮细胞,用高糖培养基干预48小时后,用免疫荧光检测Hrd1在细胞的表达变化。 结果在视网膜组织中:PCR芯片结果显示,在这89个基因中,三个月糖尿病模型组有12个基因mRNA水平发生了变化,其中Hrd1表达下降,与正常对照组相比,差异有统计学意义(P0.05);Western blot显示,三个月糖尿病模型组Hrd1蛋白质表达水平与对照组相比下降,差异有统计学意义(P0.05);免疫荧光显示, FITC标记的Hrd1在糖尿病鼠视网膜中绿色荧光减弱,表明其蛋白质表达下降,与对照组相比差异有统计学意义(P0.05)。在体外培养血管内皮细胞:免疫荧光检测结果显示,在高糖培养基干预后的细胞中绿色荧光减弱,Hrd1蛋白表达与对照组相比显著下降,差异有统计学意义(P0.05)。 结论:发生变化的12个内质网应激相关因子参与了糖尿病视网膜病变的发生和发展,Hrd1在糖尿病视网膜病变的内质网应激中起到重要作用。
[Abstract]:Objective: endoplasmic reticulum stress plays an important role in the occurrence and development of diabetic retinopathy. However, hydroxymethyl glutaryl coA reductase degrading protein 1 (Hrd1) can stabilize the environment of endoplasmic reticulum through endoplasmic reticulum associated protein degradation pathway, thus reducing the occurrence of endoplasmic reticulum stress. The changes of endoplasmic reticulum (ER) related factors and the expression of Hrd1 in diabetic rat retina and human retinal microvascular endothelial cells cultured with high glucose in vitro were analyzed by gene chip. To analyze its role in the occurrence and development of diabetic retinopathy. Methods Diabetic SD rats were induced by intraperitoneal injection of streptozotocin (STZ) (65 mg/kg) and control rats were treated with the same dose of sodium citrate solution. The blood glucose was measured by rat tail blood collection at 72 hours after injection. After 7 days blood glucose 鈮，

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