眼科新进展2018，Vol.38Issue(5)：416-420,5.DOI:10.13389/j.cnki.rao.2018.0097
高迁移率蛋白1的小干扰RNA对高糖环境下视网膜血管内皮细胞凋亡的影响
Effects of HMGB1 siRNA on the apoptosis of retinal vascular endothelial cells in high glucose environment
摘要
Abstract
Objective To investigate the effects of HMGB1 small interference RNA (siRNA) on retinal vascular endothelial cells.Methods siRNA was used to inhibit the expression of HMGB1,followed by the application of CCK8 assay,Hochest33342 staining and flow cytometry to observe the effects of HMGB1 siRNA on retinal vascular endothelial cells in high glucose environment.Meanwhile,the expression of proteins related to apoptosis was detected by Western blot.Results The transfection of HMGB1 siRNA down-regulated the protein expression level of HMGB1 by 73％ in siRNA group compared with normal control (NC) group (P ＜ 0.05),and the protein expression level of HMGB1 in siRNA group was decreased by 75％ compared with scr-siRNA group (P ＜ 0.05).There was no significant difference between NC group and scrsiRNA group (P ＞ 0.05).The total apoptotic rate of NC group,high-glucose group,scrsiRNA group and siRNA group was (0.40 ± 0.03)％,(49.80 ± 3.50)％,(47.60 ±1.98) ％ and (23.60 ± 2.40) ％ by flow cytometry.Compared with NC group,the apoptotic rates of high-glucose group,scr-siRNA group and siRNA group were increased (all P ＜ 0.05).Compared with scr-siRNA group,the apoptotic rate of HRECs in siRNA group was reduced,with significant statistical difference (P ＜ 0.05).There was no significant difference in the rate of cell apoptosis between scr-siRNA group and high-glucose group (P ＞ 0.05).Compared with the NC group,the protein expression level of cleavedcaspase3 protein in high-glucose group and scr-siRNA group were increased by (233 ±10) ％ and (266 ± 22) ％,respectively (both P ＜ 0.05);compared with scr-siRNA group,the protein expression level of cleaved-caspase 3 in siRNA group was reduced by (43 ±3) ％ (P ＜ 0.05);and there was no significant difference in the protein expression of cleaved-caspase 3 in high-glucose group and scr-siRNA group (P ＞ 0.05).Compared with the NC group,the protein expression of B-cell lymphoma-2 (Bcl-2) in high-glucose group and scr-siRNA group was decreased by (32 ± 2) ％ and (29 ± 3) ％,accordingly (both P ＜ 0.05);compared with scr-siRNA group,the protein expression level of Bcl-2 in siRNA group was increased by (42 ± 2) ％ (P ＜ 0.05);and there was no significant difference in the protein expression of Bcl-2 in high-glucose group and scr-siRNA group (P ＞ 0.05).Conclusion HMGB1 siRNA can reduce the apoptosis of retinal vascular endothelial cells in high glucose environment by inhibiting the activation of cleavedcaspase3 and increasing the expression of Bcl-2.关键词
糖尿病视网膜病变/高迁移率蛋白1/小干扰RNA/凋亡/视网膜血管内皮细胞Key words
diabetic retinopathy/high mobility protein 1/small interfering RNA/apoptosis/retinal vascular endothelial cells分类
医药卫生引用本文复制引用
郭宇婧,李宏娟,张嘉曦,付舒,姜双..高迁移率蛋白1的小干扰RNA对高糖环境下视网膜血管内皮细胞凋亡的影响[J].眼科新进展,2018,38(5):416-420,5.基金项目
国家自然科学基金项目(编号:81641057) （编号:81641057）
辽宁省科技厅计划项目(编号:201602287) （编号:201602287）
辽宁省大学生创新创业课题(编号:2015101600) （编号:2015101600）
锦州医科大学校长基金(编号:XZJJ20130213)National Natural Science Foundation of China (No:81641057) （编号:XZJJ20130213）
Science and Technology Project of Liaoning Province (No:201602287) （No:201602287）
Graduate Innovation Foundation of Liaoning Province (No:2015101600) （No:2015101600）
President Fund of Jinzhou Medical University (No:XZJJ20130213) （No:XZJJ20130213）
