miR-132对缺氧/复氧条件下人视网膜微血管内皮细胞中血管新生相关基因表达的影响

Effect of microRNA-132 on Gene Expression Related to Retinal Neovascularization(RNV)in Human Retinal Microvascular Endothelial Cells(HRMECs)under Hypoxia/Reoxygenation(H/R)

【目的】探索miR-132对缺氧/复氧(H/R)条件下的人视网膜微血管内皮细胞(HRMECs)中视网膜血管新生(RNV)相关的基因表达的影响。【方法】HRMECs细胞株在缺氧条件下培养6 h,然后在常氧条件下培养6 h,作为H/R组;在H/R条件下,添加miR-132的抑拮抗剂(anti-miR-132)作为H/R+anti-miR-132组;常氧条件下培养HRMECs作为阴性对照组。提取细胞总mRNA,在mRNA微阵列上检测基因表达模式的变化,使用软件包Rosetta Resolver 7.2进行Pathway分析,最后结合Pubmed数据库确定与RNV相关的候选基因。【结果】与阴性对照组比较,H/R组中细胞黏附、脂肪细胞因子、细胞外基质受体相互作用等细胞信号变化显著(P<0.05);通过抑制miR-132的表达,发现miR-132参与到H/R对细胞中多种信号分子的调控;结合Pubmed数据库的分析,筛选出三个与RNV相关的基因:THBS1,IL-6和CXCL8。【结论】miR-132通过调控多个基因的表达参与H/R诱导的RNV,这有助于进一步阐明H/R诱导RNV发生的机制。

【Objective】To explore the effect of microRNA-132 on gene expression related to retinal neovascularization(RNV)in human retinal microvascular endothelial cells(HRMECs)under hypoxia/reoxygenation(H/R).【Methods】HRMECs cell lines were cultured under hypoxia for 6 hours and then under normal oxygen for 6 hours as H/R group;anti-mir-132 was added as H/R+anti-mir-132 group under H/R condition;HRMECs were cultured under normal oxygen condition as negative control group.Total cell RNA was extracted and the changes of gene expression patterns were detected on the RNA microarray.Pathway analysis was performed using Rosetta Resolver 7.2 software package.Finally,candidate genes related to RNV were identified with Pubmed database.【Results】Compared with the negative control group,there were significant changes in cell signals such as cell adhesion,adipocytokine and extracellular matrix receptor interaction in the H/R group(P<0.05).By inhibiting the expression of microRNA-132,we found that microRNA-132 participated in the regulation of various signal molecules in H/R cells.Three RNV related genes were screened out based on the analysis of Pubmed database:THBS1,IL-6 and CXCL8.【Conclusion】Mir-132 participates in H/R-induced RNV by regulating the expression of multiple genes,which helps to further elucidate the mechanism of H/R-induced RNV.