SC79激活视网膜色素上皮细胞Akt-Nrf2信号通路抗紫外线氧化损伤

发布时间：2018-04-18 18:58

本文选题：SC79 + 视网膜色素上皮(RPE)细胞　；参考：《南京医科大学》2017年博士论文

【摘要】：研究目的:过度的紫外线导致视网膜色素上皮氧化应激和细胞凋亡。我们研究视网膜色素上皮细胞中,新型Akt激动剂SC79抗紫外线(UV)氧化损伤作用,重点解析其相关分子机制。研究方法:为检测SC79对UV引起的RPE细胞的保护作用,我们使用MTT、台盼蓝染色检测细胞活性;流式细胞仪、组蛋白DNAELISA、Caspase-9活性以及线粒体去极化检测等方法检测细胞凋亡。我们使用Western blot检测SC79活化RPE细胞中的Akt水平。通过Akt抑制剂MK-2206、shRNA沉默Aktl基因,分析Akt信号通路机制;通过qRT-PCR检测RPE细胞中HO-1、NQO1、Nrf2变化的影响,以及Nrf2shRNA、显性负性突变的Nrf2(DN,S40T)对SC79保护作用的影响,分析Nrf2信号通路机制。小鼠玻璃体腔注射SC79,通过视觉电生理检测其对视网膜光损伤的保护作用。研究结果:SC79保护RPE细胞免受UV损害。SC79激活Akt,抑制UV引起的RPE细胞损伤;SC79活化Akt下游Nrf2信号,保护RPE细胞免受UV诱导氧化损伤。SC79玻璃体腔内注射减轻小鼠视网膜光损害。结论:SC79激活RPE细胞Akt-Nrf2信号通路,抑制UV氧化损伤。
[Abstract]:Objective: excessive ultraviolet radiation induced oxidative stress and apoptosis of retinal pigment epithelium.We studied the effects of a novel Akt agonist, SC79, on the oxidative damage of retinal pigment epithelial cells (RPE), focusing on its molecular mechanisms.Methods: in order to detect the protective effect of SC79 on UV-induced RPE cells, we used MTTand Trypan blue staining to detect cell activity, flow cytometry, histone DNA ELISA-Caspase-9 activity and mitochondrial depolarization to detect apoptosis.We used Western blot to detect the level of Akt in RPE cells activated by SC79.The mechanism of Akt signaling pathway was analyzed by Akt inhibitor MK-2206 siRNA silencing Aktl gene, and the changes of HO-1NQO1Nrf2 in RPE cells were detected by qRT-PCR, and the protective effect of Nrf2shRNAand Nrf2DNN S40T on SC79 was analyzed, and the mechanism of Nrf2 signaling pathway was analyzed.SC79 was injected into the vitreous cavity of mice and the protective effect of SC79 on retinal light injury was detected by visual electrophysiology.Results RPE cells were protected from UV damage. SC79 activated Akt.SC79 inhibited UV-induced injury of RPE cells and activated downstream Nrf2 signal of Akt. It also protected RPE cells from UV-induced oxidative damage. SC79 intravitreal injection alleviated retinal light damage in mice.Conclusion the Akt-Nrf2 signaling pathway of RPE cells was activated by WSC79 and UV oxidative damage was inhibited.
【学位授予单位】：南京医科大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R774

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