miR-155-5p靶向调控CCN1对高糖诱导的人视网膜血管内皮细胞增殖、凋亡和迁移的影响OA

Objective To investigate the effect of microRNA-155-5p(miR-155-5p)on high glucose-induced prolifera-tion,apoptosis,and migration of human retinal vascular endothelial cells(HRECs)through targeted regulation of cysteine-rich vascular inducing factor 61(CCN1).Methods HRECs were divided into the negative control(NC)group,high glu-cose(HG)group,miR-155-5p agonist negative control(agomir-NC)group,miR-155-5p agonist(miR-155-5p-agomir)group,miR-155-5p-agomir+empty plasmid(pc-NC)group,and miR-155-5p-agomir+CCN1 overexpression plasmid(pc-CCN1)group.Real-time fluorescence quantitative polymerase chain reaction(RT-PCR)was performed to detect the mRNA expression levels of miR-155-5p and CCN1 in cells.Cell proliferation was detected by methyl thiazolyl tetrazolium(MTT)assay.Cell migration was assessed by a wound healing assay.Flow cytometry was used to detect cell apoptosis.An in vitro tube formation assay was performed to assess the tube forming ability of cells.Western blot was used to detect the protein expression levels of matrix metalloproteinase-2(MMP-2),cleaved cysteinyl aspartate-specific proteinase 3(cleaved Caspase-3),proliferating cell nuclear antigen(PCNA),vascular endothelial growth factor(VEGF),and CCN1 in the cells.The relationship between miR-155-5p and CCN1 was validated.Results Compared with the HG and agomir-NC groups,the miR-155-5p-agomir group showed decreased CCN1 mRNA expression level,OD490(24,48 hours)values,wound healing rate,and apoptosis rate in HRECs,along with a reduced number of tube formations;besides,the relative protein expression levels of MMP-2,cleaved Caspase-3,PCNA,VEGF,and CCN1 decreased,while the relative expression level of miR-155-5p in-creased.All these differences were statistically significant(all P<0.05).Compared with the miR-155-5p-agomir+pc-NC group,the miR-155-5p-agomir+pc-CCN1 group exhibited increased CCN1 mRNA expression level,OD490(24,48 hours)val-ues,wound healing rate,and apoptosis rate in HRECs,along with an enhanced number of tube formations;in addition,the relative protein expression levels of MMP-2,cleaved Caspase-3,PCNA,VEGF,and CCN1 increased,and all these differences were statistically significant(all P<0.05).The luciferase activity in the miR-155-5p mimics+CCN1-WT group decreased compared with the miR-NC+CCN1-WT group(P<0.05).Conclusion miR-155-5p can inhibit high glucose-induced proliferation,apoptosis,migration,and angiogenesis of HRECs,which may be related to targeted regulation of CCN1.