miR-1-3p调控ANXA2表达对高糖诱导的视网膜微血管内皮细胞新生血管生成的影响  

Effect of micro ribonucleic acid-1-3p on high glucose-induced neovascularization of retinal microvascular endothelial cells through regulation of Annexin A2 expression

作　　者：巨朝娟[1] 许寅聪 李康宁[1] 石笑楠 熊朝晖 戴明明 JU Chaojuan;XU Yincong;LI Kangning;SHI Xiaonan;XIONG Zhaohui;DAI Mingming(Department of Ophthalmology,the First Hospital of Hebei Medical University,Shijiazhuang 050031,Hebei Province,China)

机构地区：[1]河北医科大学第一医院眼科,河北省石家庄市050031

出　　处：《眼科新进展》2023年第12期952-957,共6页Recent Advances in Ophthalmology

基　　金：河北省重点科技研究计划(编号:20210842);河北省卫生健康委员会项目(编号:20191762)。

摘　　要：目的 探讨微小RNA-1-3p(miR-1-3p)/膜联蛋白A2(ANXA2)分子轴在高糖诱导的人视网膜微血管内皮细胞(HRMECs)新生血管生成过程中的作用机制。方法 体外培养HRMECs并采用高糖(HG)处理细胞建立细胞损伤模型。HRMECs分组处理:Con组(含体积分数10%胎牛血清的DMEM培养)、HG组(25 mmol·L^(-1)D-葡萄糖培养)、HG+miR-NC组(转染miR-NC)、HG+miR-1-3p组(转染miR-1-3p mimics)、HG+sh-NC组(转染sh-NC)、HG+sh-ANXA2组(转染sh-ANXA2)、HG+miR-1-3p+pcDNA组(转染miR-1-3p mimics+pcDNA)、HG+miR-1-3p+pcDNA-ANXA2组(转染miR-1-3p mimics+pcDNA-ANXA2)。转染48 h后收集细胞,采用25 mmol·L^(-1)的D-葡萄糖培养基培养HRMECs 24 h。采用MTT、Transwell小室实验分别检测细胞活力、迁移细胞数。管腔形成实验检测管腔形成数。双荧光素酶报告实验检测miR-1-3p与ANXA2的靶向关系。Western blot检测VEGF、MMP-2蛋白水平。结果 与Con组比较,HG组miR-1-3p表达水平降低,ANXA2 mRNA及蛋白水平均升高,差异均有统计学意义(均为P<0.05)。与Con组比较,HG组细胞活力升高,迁移细胞数、管腔形成数增多,VEGF、MMP-2蛋白水平升高,差异均有统计学意义(均为P<0.05)。与HG+miR-NC组比较,HG+miR-1-3p组细胞活力降低,迁移细胞数、管腔形成数减少,VEGF、MMP-2蛋白水平降低,差异均有统计学意义(均为P<0.05)。与HG+sh-NC组比较,HG+sh-ANXA2组细胞活力降低,迁移细胞数、管腔形成数减少,VEGF、MMP-2蛋白水平降低,差异均有统计学意义(均为P<0.05)。与HG+miR-1-3p+pcDNA组比较,HG+miR-1-3p+pcDNA-ANXA2组细胞活力升高,迁移细胞数、管腔形成数增多,VEGF、MMP-2蛋白水平升高,差异均有统计学意义(均为P<0.05)。结论 miR-1-3p过表达可通过靶向调控ANXA2表达抑制HRMECs的增殖、迁移及新生血管生成。Objective To investigate the possible action mechanism of the micro ribonucleic acid-1-3p(miR-1-3p)/Annexin A2(AnxA2)molecular axis in high glucose(HG)-induced neovascularization of human retinal microvascular endothelial cells(HRMECs).Methods A cell injury model was established by culturing HRMECs in vitro and treating them with HG.The HRMECs were divided into the Con group(DMEM medium containing fetal bovine serum in volume fraction of 10%),HG group(cultured in 25 mmol·L^(-1) D-glucose),HG+miR-NC group(transfected with miR-NC),HG+miR-1-3p group(transfected with miR-1-3p mimics),HG+sh-NC group(transfected with sh-NC),HG+sh-AnxA2 group(transfected with sh-AnxA2),HG+miR-1-3p+pcDNA group(transfected with miR-1-3p mimics+pcDNA),and HG+miR-1-3p+pcDNA-AnxA2 group(transfected with miR-1-3p mimics+pcDNA-AnxA2).After 48 h of transfection,cells were collected and cultured in 25 mmol·L^(-1) D-glucose medium for 24 h.Cell viability and number of migrating cells were detected using MTT and Transwell chamber experiments,respectively.The number of lumen formations was detected by the lumen formation experiment.The dual luciferase reporter assay was adopted to detect the targeting relationship between miR-1-3p and AnxA2.Western blot was used to detect the protein levels of vascular endothelial growth factor(VEGF)and matrix metalloproteinase-2(MMP-2).Results Compared with the Con group,the expression level of miR-1-3p in the HG group decreased,while the levels of AnxA2 messenger ribonucleic acid(mRNA)and protein increased,with statistically significant differences(all P<0.05).Compared with the Con group,the HG group showed an increase in cell viability,number of migrating cells,lumen formation and protein levels of VEGF and MMP-2,with statistically significant differences(all P<0.05).Compared with the HG+miR-NC group,the HG+miR-1-3p group showed a decrease in cell viability,number of migrating cells,lumen formation and protein levels of VEGF and MMP-2,with statistically significant differences(all P<0.05).Compared with the HG+

关 键 词：微小RNA-1-3p 膜联蛋白A2 高糖 视网膜微血管内皮细胞 新生血管 细胞增殖 细胞迁移 

分 类 号：R774.1[医药卫生—眼科]