抑制PTEN对视网膜脱离后光感受器细胞的保护作用

发布时间：2018-02-26 10:55

本文关键词： PTEN 视网膜脱离光感受器细胞凋亡 PI3K/Akt信号通路　出处：《上海交通大学》2015年博士论文　论文类型：学位论文

【摘要】：目的:视网膜脱离后光感受器细胞的凋亡是视功能恢复不良的的主要原因。PTEN是神经元凋亡发生中关键的调节因素。PTEN/PI3K/Akt信号通路作为一种细胞存活信号通路,其神经保护作用已得到确认,而且它的作用靶点也逐渐得到确定。本课题采用PTEN的特异性抑制剂bp V(pic)干预,拟研究PTEN/PI3K/Akt信号通路及其下游分子以及光感受器细胞凋亡的变化特点,阐明PTEN/PI3K/Akt信号通路对视网膜脱离后光感受器细胞凋亡的调控机制。方法:建立视网膜脱离模型,成模后1 d、2 d、3 d和7 d运用Western Blot和免疫荧光检测视网膜内PTEN表达的变化。建模后视网膜下注射bp V(pic),运用Western Blot和免疫荧光检测p-Akt和Bcl-2在视网膜的表达与分布;运用Western Blot检测视网膜内p-PDK1、p-BAD、胞浆Cyt c和Caspase-3蛋白水平的表达;ELISA检测PIP3的表达水平;运用TUNEL检测视网膜的细胞凋亡情况;记录ONL的厚度。结果:视网膜脱离后PTEN的表达逐步上调,3 d为峰值(P0.05),之后逐渐下降,7 d时基本恢复到正常。视网膜下注射bp V(pic)后,能使视网膜脱离后光感受器细胞的凋亡显著减少(P0.05),视网膜ONL厚度增加(P0.05)。与vehicle-treated RD组相比,bp V-treated RD组的p-Akt和p-PDK1显著升高(P0.05)。此外,bp V还能使p-BAD和Bcl-2的表达上调(P0.05),胞浆Cyt c和Caspase-3的表达降低(P0.05)。结论:PTEN参与了视网膜脱离后光感受器细胞的凋亡过程。抑制PTEN可通过激活PI3K/Akt信号通路,抑制线粒体通路的活化,从而减少光感受器细胞凋亡的发生。
[Abstract]:Objective: apoptosis of photoreceptor cells after retinal detachment is the main cause of poor recovery of visual function. PTEN is a key regulatory factor in neuronal apoptosis. PTEN / PI3K / Akt signaling pathway is a cell survival signal pathway. Its neuroprotective effect has been confirmed, and its target has been gradually determined. In this study, the characteristics of apoptosis of PTEN/PI3K/Akt signaling pathway, its downstream molecules and photoreceptor cells were studied with the intervention of BP Vpica, a specific inhibitor of PTEN. To elucidate the mechanism of PTEN/PI3K/Akt signaling pathway regulating photoreceptor cell apoptosis after retinal detachment. Methods: retinal detachment model was established. The changes of PTEN expression in the retina were detected by Western Blot and immunofluorescence on the 3rd and 7th day after the model was established. The expression and distribution of p-Akt and Bcl-2 in the retina were detected by Western Blot and immunofluorescence, and the expression and distribution of p-Akt and Bcl-2 in the retina were detected by Western Blot and immunofluorescence. Western Blot was used to detect p-PDK1 p-BAD. the expression of cytoplasmic Cyt c and Caspase-3 protein was detected by Elisa, and TUNEL was used to detect the apoptosis of retina. The thickness of ONL was recorded. Results: after retinal detachment, the expression of PTEN was gradually up-regulated to a peak value of P0.05 at 3 days, then gradually decreased to normal at 7 days. Compared with vehicle-treated Rd group, p-Akt and p-PDK1 were significantly increased in p-Akt and p-PDK1 in vehicle-treated Rd group. In addition, BPV could also up-regulate the expression of p-BAD and Bcl-2, and increase the expression of p-BAD and Bcl-2 in the cytoplasm Cyt c. Conclusion Caspase-3 is involved in the apoptosis of photoreceptor cells after retinal detachment. Inhibition of PTEN may be mediated by activation of PI3K/Akt signaling pathway. Inhibition of mitochondrial pathway activation, thereby reducing photoreceptor cell apoptosis.
【学位授予单位】：上海交通大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R774.12

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