miR-23a通过靶向GLS1调控H_(2)O_(2)诱导的视网膜色素上皮细胞增殖、迁移  

miR-23a regulates HO-induced proliferation and migration of retinal pigment epithelial cells via targeting GLS1

作　　者：田蕴霖 周歆 白淑玮[1] 姚倩[1] TIAN Yun-lin;ZHOU Xin;BAI Shu-wei;YAO Qian(Ophthalmology Department of the Fourth Hospital of Xi’an,Xi'an 710004,China)

机构地区：[1]西安市第四医院眼科,陕西西安710004

出　　处：《解剖科学进展》2021年第1期119-124,128,共7页Progress of Anatomical Sciences

基　　金：国家自然科学基金(81302198)。

摘　　要：目的探究氧化应激条件下miR-23a对视网膜色素上皮细胞增殖、迁移的影响及其机制。方法以不同浓度的H_(2)O_(2)诱导ARPE-19细胞氧化应激,qPCR检测诱导后ARPE-19细胞miR-23a的表达水平以及增殖和迁移能力的变化;ARPE-19细胞转染miR-23a mimic、NC(negative control) mimic后,采用MTT和Transwell分别检测各组细胞的增殖和迁移能力;双荧光素酶报告实验验证miR-23a和GLS1的靶向关系;检测GLS1表达对氧化应激下ARPE-19细胞增殖和迁移能力的影响;检测转染miR-23a mimic及调节GLS1表达后ARPE-19细胞谷氨酰胺摄取能力,Western blot检测转染miR-23a mimic及调节GLS1表达后ARPE-19细胞p42/p44 MAPK、JNK磷酸化水平。结果 H_(2)O_(2)诱导后ARPE-19细胞miR-23a表达水平显著上调,细胞增殖和迁移能力显著下降;转染miR-23a mimic后ARPE-19细胞增殖和迁移能力显著抑制。荧光素酶报告实验显示GLS1为miR-23a的靶基因,下调GLS1表达后,显著抑制H_(2)O_(2)诱导的ARPE-19细胞增殖和迁移能力;上调miR-23a及下调GLS1的表达则抑制H_(2)O_(2)诱导ARPE-19细胞的谷氨酰胺的摄取及p42/p44 MAPK、JNK磷酸化水平。结论氧化应激通过上调视网膜色素上皮细胞miR-23a而抑制GLS1的表达进而抑制MAPK信号通路影响细胞的增殖和迁移。Objective To investigate the effect and possible mechanism of miR-23 a on the proliferation and migration of retinal pigment epithelial cells under oxidative stress. Methods The oxidative stress of ARPE-19 cells was induced with different concentrations of H_(2)O_(2). QPCR was used to detect the expression of miR-23a. ARPE-19 cells were divided into: miR-23a mimic, NC(negative control) mimic, the cell proliferation and migration ability were determined by MTT and transwell respectively. The double luciferase report experiment was used to verify the targeting relationship between miR-23a and GLS1;the expressions of GLS1, p42/p44 MAPK, JNK phosphorylation in ARPE-19 cells were detected by Western blot. Results After H_(2)O_(2) induction, the expression level of miR-23a in ARPE-19 cells was significantly up-regulated, and the cell proliferation and migration ability were significantly decreased. After transfection with miR-23a mimic, ARPE-19 cell proliferation and migration abilities were significantly inhibited. The luciferase report experiment showed that GLS1 was a target gene of miR-23a, down-regulating the expression level of GLS1 inhibited the proliferation and migration of H_(2)O_(2)-induced ARPE-19 cells, and up-regulated miR-23a and down-regulated GLS1 expression inhibited glutamine uptake and p42/p44 MAPK and JNK phosphorylation levels in ARPE-19 cells after H_(2)O_(2) induction. Conclusion Oxidative stress inhibited the expression of GLS1 via up-regulating miR-23a in retinal pigment epithelial cells, and then inhibited MAPK signaling pathway to affect cell proliferation and migration.

关 键 词：氧化应激 视网膜色素上皮细胞 增殖 迁移 机制 

分 类 号：R774[医药卫生—眼科]