【摘要】 目的：探讨E2F1对高糖诱导的视网膜神经节细胞(retinal ganglion cells,RGCs)存活和凋亡的影响及其作用机制。方法：以RGC-5细胞为研究对象，构建高糖损伤模型，将RGC-5细胞转染后分为空白对照组、高糖组、高糖+si-E2F1组、高糖+si-con组、高糖+si-E2F1+PBS组、高糖+si-E2F1+SD19组。分别采用实时荧光定量PCR(real-time RT-PCR)与蛋白质印迹法检测细胞中E2F1 mRNA表达；四甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)检测细胞存活率，TUNEL法和流式细胞术(flowcytometry,FCM)检测细胞凋亡。蛋白质印迹法检测细胞中E2F1、磷酸化的转录激活子3(p-signal transducer and activator of transcription 3, p-STAT3)、 STAT3、磷酸化的Janus蛋白酪氨酸激酶2(p-Januskinase2,p-JAK2)、JAK2、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(cleavedcaspase-3)和活化的多聚二磷酸腺苷核糖聚合酶(cleaved poly-ADP-ribose polymerase,cleaved PARP)蛋白表达。结果：与空白对照组相比，高糖组RGC-5细胞中E2F1表达水平升高，细胞存活率明显下降，p-STAT3、p-JAK2、cleavedcaspase-3、cleavedPARP、E2F1表达及细胞凋亡率显著升高；沉默E2F1表达后，细胞存活率明显升高，而cleaved caspase-3蛋白和cleaved PARP蛋白表达及细胞凋亡率降低。SD19可部分逆转沉默E2F1对RGC-5细胞的保护作用。结论：抑制E2F1表达能够减轻高糖诱导的RGCs凋亡，其机制可能与抑制JAK2/STAT3信号通路有关。更多还原

【Abstract】 Objective: To investigate the effect of E2F1 on the survival and apoptosis of retinal ganglion cells(RGCs) induced by high glucose and its mechanism. Methods: A high-glycemic injury model was constructed with.RGC-5 cells used as the research object. After transfection, RGC-5 cells were divided into a blank control group, a high-sugar group, a high-sugar + si-E2F1 group, a high-sugar + si-con group, a high glucose + si-E2F1 + PBS group, and a high glucose + si-E2F1 + SD19 group. Real-time RT-PCR and Western blotting were used to detect the expression of E2F1 m RNA in cells; methyl thiazolyl tetrazolium(MTT) was used to detect cell viability; TUNEL and flow cytometry(FCM) were used to detect cell apoptosis. E2F1, p-signal transducer and activator of transcription 3(p-STAT3), STAT3, p-Janus kinase 2(p-JAK2), JAK2, cleaved caspase-3, and cleaved poly-ADPribose polymerase(cleaved PARP) protein expression in cells detected by Western blotting. Results: Compared with blank control group, E2F1 expression level increased in RGC-5 cells in the high glucose group, cell survival rate was significantly decreased, p-STAT3, p-JAK2, cleaved caspase-3, cleaved PARP, E2F1 expression, and apoptotic rate were increased. After silencing E2F1 expression, the cell survival rate was significantly increased, while cleaved caspase-3 protein expression, cleaved PARP protein expression, and apoptotic rate were decreased. SD19 partially reversed the protective effect of silencing E2F1 on RGC-5 cells. Conclusion: Inhibition of E2F1 expression can reduce high glucose-induced apoptosis in RGCs. The mechanism may be related to the inhibition of the JAK2/STAT3 signaling pathway.更多还原