摘要
目的 探讨miR-495通过靶向 Notch1信号调控高糖诱导的视神经节细胞(RGCs)凋亡的效果与机制。方法 将高糖培养的RGC-5细胞系随机分为三组:空白组、miR-495阴性对照(miR NC)和miR-495组。miR-NC组与miR-495组分别转染miR-495模拟物阴性对照(miR NC)与miR-495 模拟物(mimic),空白组不进行转染。采用qRT-PCR检测miR-495表达,CCK-8法检测细胞增殖指数,流式细胞术检测检测细胞周期,流式细胞术检测细胞凋亡指数,Western Blot检测Notch1蛋白表达。结果 转染后24 h与36 h,细胞增殖指数显著高于空白组与miR-NC组( P <0.05);细胞凋亡指数显著低于空白组与miR-NC组( P <0.05);转染后36 h,miR-495组的G1期比率相对于miR-NC组和空白组显著降低( P <0.05),S期+G2期比率则显著增加( P <0.05)。转染后24 h与36 h,miR-495组的Notch1蛋白相对表达量显著低于miR-NC组和空白组,空白组与miR-NC组比较差异无统计学意义( P >0.05)。结论 miR-495能减轻高糖诱导的RGCs损伤,促进RGCs增殖,抑制Notch1信号通路,从而调节细胞周期,抑制RGCs凋亡。
Objective To investigate the effect and mechanism of miR-495 on the apoptosis of retinal ganglion cells (RGCs) induced by high glucose by targeting Notch1 signaling. Methods High glucose cultured RGC-5 cells were randomly divided into three groups, including blank group, miR-NC group and miR-495 group. miR-NC group and miR-495 group were transfected with miR NC and miR-495 mimic, respectively. The blank group were not transfected. The expression of miR-495 were detected by qRT-PCR, the cell proliferation index were detected by CCK-8 method, the cell cycle were detected by flow cytometry, the apoptosis index were detected by flow cytometry, and the expression of Notch1 protein were detected by Western Blot. Results At 24 h and 36 h after transfection, the cell proliferation index in the miR-495 group were significantly higher than that of the blank group and the miR-NC group ( P <0.05), and the apoptotic index were significantly lower than that of the blank group and the miR-NC group ( P <0.05). At 36 h, the proportion of G1 in the miR-495 group were significantly lower than that in the miR-NC group and the blank group ( P <0.05), and the proportion in the S+G2 phase were significantly increased ( P <0.05). At 24 h and 36 h after transfection, the relative expression of Notch1 protein in miR-495 group were significantly lower than that in miR-NC group and blank group. There were no significant difference compared between blank group and miR-NC group ( P >0.05). Conclusion miR-495 can attenuate the damage of RGCs induced by high glucose, promote the proliferation of RGCs, inhibit the signaling pathway of Notch1, regulate cell cycle and inhibit the apoptosis of RGCs.
作者
杨晓
张胜利
赵俊宏
张秀丽
刘钊
YANG Xiao;ZHANG Sheng-li;ZHAO Jun-hong(Department of Ophthalmology, Weinan First Hospital, Weinan Shaanxi 714000, China;Department of Ophthalmology, Xi'an First Hospital, Xi'an Shaanxi 710002, China)
出处
《临床和实验医学杂志》
2019年第14期1486-1489,共4页
Journal of Clinical and Experimental Medicine
基金
陕西省科学技术厅软科学研究计划一般项目(编号:2018KRM174)
