miR-135a-5p通过调控Bcl-2/Bax通路对青光眼视网膜神经节细胞凋亡的作用机制  

Mechanism of miR-135a-5p on apoptosis of retinal ganglion cells in glaucoma by regulating Bcl-2/Bax pathway

作　　者：谢九冰 杨杉杉[1] 陈喜月[1] 岳向东[1] XIE Jiubing;YANG Shanshan;CHEN Xiyue;YUE Xiangdong(Department of Glaucoma,Tangshan Eye Hospital,Tangshan,Hebei 063000,China)

机构地区：[1]河北省唐山市眼科医院青光眼科,063000

出　　处：《重庆医学》2022年第8期1290-1296,共7页Chongqing medicine

基　　金：河北省卫生厅科研基金项目(20201548)。

摘　　要：目的探讨miR-135a-5p与Bcl-2/Bax间的关系及其对青光眼视网膜神经节细胞(RGCs)凋亡的作用和机制。方法建立BALB/C小鼠青光眼模型,检测眼压和视网膜RGCs数量及miR-135a-5p表达。实验用RGCs细胞分为miR-135a-5p组、空白组、miR-135a-5p抑制组,miR-135a-5p组转染miR-135a-5p mimic,空白组转染无意义序列miRNA-NC,miR-135a-5p抑制组转染si-miR-135a-5p。qRT-PCR检测miR-135a-5p表达水平,流式细胞术检测各组RGCs细胞凋亡率,CCK-8检测各组RGCs细胞活性,Western blot测定各组RGCs中Bcl-2、Bax蛋白表达。结果随着小鼠青光眼模型作用时间增加,小鼠视网膜内miR-135a-5p表达逐渐降低,RGCs活细胞数量逐渐降低(P<0.05);miR-135a-5p组miR-135a-5p表达水平及RGCs活性均高于空白组,空白组高于miR-135a-5p抑制剂组(P<0.05);miR-135a-5p组RGCs Bcl-2蛋白表达高于空白组,空白组高于miR-93-5p抑制剂组(P<0.05);miR-135a-5p组RGCs Bax蛋白表达水平低于空白组,空白组低于miR-135a-5p抑制剂组(P<0.05);miR-135a-5p组RGCs凋亡率低于空白组,空白组低于miR-135a-5p抑制剂组(P<0.05)。结论miR-135a-5p在青光眼RGCs中呈低表达,miR-135a-5p高表达能降低青光眼RGCs的凋亡,其机制可能是通过调控miR-135a-5p/Bcl-2/Bax轴实现。Objective To investigate the relationship between miR-135a-5p and Bcl-2/Bax and its effect and mechanism of miR-135a-5p on apoptosis of retinal ganglion cells(RGCs)in glaucoma.Methods Glaucoma in BALB/C rats′models were established,and the intraocular pressure,RGCs quantity and miR-135a-5p expression level were measured.RGCs cells were divided into the miR-135a-5p group,the blank group and the miR-135a-5p inhibition group.The miR-135a-5p group was transfected with miR-135a-5p mimic,the blank group was transfected with meaningless sequence(miRNA-NC),and the miR-135a-5p inhibition group was transfected with si-miR-135a-5p.The expression level of miR-135a-5p was detected by qRT-PCR,the apoptosis rate of RGCs cells in each group was detected by flow cytometry,CCK-8 was used to detect the cell activity,and Western blot was used to detect the expression of Bcl-2 and Bax protein in RGCs.Results The expression level of miR-135a-5p gradually decreased and the number in living cells decreased with the increase of the time of glaucoma in rats′models(P<0.05).The expression of miR-135a-5p and the amount of RGCs in the miR-135a-5p group were higher than those in the blank group,and those in the blank group were higher than those in the miR-135a-5p inhibitor group(P<0.05).The expression of Bcl-2 protein in RGCs in the miR-135a-5p group was higher than that in the blank group,and that in the blank group was higher than that in the miR-93-5p inhibitor group(P<0.05).The expression of Bax protein in the miR-135a-5p group was lower than that in the blank group,and that in the blank group was lower than that in the miR-135a-5p inhibitor group(P<0.05).The apoptosis rate of RGCs in the miR-135a-5p group was lower than that in the blank group,and that in the blank group was lower than that in the miR-135a-5p inhibitor group(P<0.05).Conclusion The expression of miR-135a-5p is decreased in RGCs of glaucoma,and the high expression of miR-135a-5pp can reduce the apoptosis of RGCs of glaucoma,which may be realized by regulating miR-135a-

关 键 词：青光眼 神经节细胞 miR-135a-5p Bcl-2 BAX 细胞凋亡 

分 类 号：R775[医药卫生—眼科]