ERAD相关因子在糖尿病鼠视网膜中的表达变化

发布时间：2018-07-04 23:40

本文选题：内质网相关降解 + 糖尿病视网膜病变　；参考：《华中科技大学》2011年硕士论文

【摘要】：目的:内质网相关降解(ER-associated degradation, ERAD)是内质网中重要的蛋白质质量监控系统,能够降解错误折叠的蛋白,避免细胞发生内质网应激,内质网应激与糖尿病视网膜病变的发生、发展有密切关系。本研究通过建立SD大鼠糖尿病模型,检测ERAD作用途径相关因子在糖尿病鼠视网膜中的表达变化,从而探讨ERAD相关因子在糖尿病视网膜病变的早期阶段的发生和发展中可能存在的作用。方法:用大剂量链链脲佐菌素(Streptozotocin ,STZ)一次性腹腔注射造成糖尿病SD大鼠模型,以同龄SD大鼠腹腔注射等量的柠檬酸钠缓冲液为对照组。自行设计实时定量PCR芯片检测内质网应激相关因子在一月糖尿病鼠视网膜中的表达变化,Western blot、免疫组化检测一月糖尿病鼠视网膜中ERAD相关因子Herp蛋白的表达变化。体外培养人视网膜微血管内皮细胞,高糖干预后,免疫荧光检测Herp蛋白的表达变化。结果:实时定量PCR芯片结果显示:一月糖尿病鼠视网膜中有13个内质网应激相关因子的mRNA水平明显下调(P0.05),涵盖内质网应激中了8个作用途径,其中ERAD相关因子Herp, NUC, Os9, ADO15的表达明显下调(P0.05)。Western blot与免疫荧光提示,Herp蛋白在一月糖尿病模型鼠视网膜中的表达明显下调(P0.05);体外培养人视网膜微血管内皮细胞在高糖干预后,免疫荧光显示Herp在蛋白水平的表达明显下调(P0.05)。结论:内质网应激与糖尿病视网膜病变的发生与发展有密切的关系, ERAD作用途径参与了糖尿病视网膜病变的发生和发展,Herp在该过程中发挥了重要的的作用。
[Abstract]:Objective: ER-associated degradation (ERAD) is an important protein quality monitoring system in the endoplasmic reticulum (ER), which can degrade misfolded proteins and avoid endoplasmic reticulum stress, endoplasmic reticulum stress and diabetic retinopathy. Development is closely related. In this study, SD rat model of diabetes mellitus was established to detect the expression of ERAD pathway related factors in the retina of diabetic rats. To explore the role of ERAD related factors in the early stage of diabetic retinopathy. Methods: a diabetic SD rat model was established by intraperitoneal injection of streptozotocin (STZ) in a single dose, and sodium citrate buffer was injected intraperitoneally to the same age SD rats as control group. A real-time quantitative PCR chip was designed to detect the expression of ER stress-related factors in the retina of diabetic rats in January. The expression of ERAD-related factor Herp protein in the retina of diabetic rats in January was detected by immunohistochemistry. Human retinal microvascular endothelial cells were cultured in vitro. The expression of Herp protein was detected by immunofluorescence after high glucose intervention. Results: the results of real-time quantitative PCR microarray showed that the mRNA levels of 13 ER stress-related factors in the retina of diabetic rats were significantly down-regulated (P0.05), covering 8 pathways of endoplasmic reticulum stress. The expression of Herp, NUC, Os9, ADO15 was significantly down-regulated (P0.05). Western blot and immunofluorescence indicated that Herp protein expression was significantly down-regulated in the retina of diabetic rats in January (P0.05); in vitro cultured human retinal microvascular endothelial cells (RMECs) were treated with high glucose. Immunofluorescence showed that Herp expression was significantly down-regulated at protein level (P0.05). Conclusion: endoplasmic reticulum stress is closely related to the occurrence and development of diabetic retinopathy. ERAD plays an important role in the pathogenesis and development of diabetic retinopathy.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R774.1

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