目的:观察H2 O2处理对晶状体上皮细胞HLE-B3miRNA表达谱的影响,初步明确miRNA在氧化损伤诱导晶状体细胞凋亡中的作用及机制。方法:100μmol/L H2 O2处理HLE-B324h后,收集细胞用Trizol法提取总RNA。使用芯片检测miRNA的表达,并且qRT-PCR对芯片结果进行验证。运用生物信息学方法预测差异miRNA调控的靶基因。结果:HLE-B3细胞经H2 O2处理后,28个miRNA表达发生明显变化(18个上调,10个下调)。差异miRNA可能通过调控BCL2 L2和MIP的表达,参与了晶状体细胞凋亡,进而导致了白内障的发生发展。结论:氧化损伤能够明显改变晶状体上皮细胞的miRNA表达谱,此变化可能在白内障的发生发展中起调节作用。
AIM: To identify the changes in microRNA ( miRNA ) profile of human lens epithelium ( HLE) induced by H2 O2 and the role of miRNA in oxidative stress induced apoptosis. METHODS: HLE cell line HLE - B3 was treated by 100μmol/L H2 O2 for 24h and the total RNA were isolated by Trizol reagent. miRNA profile was generated by miRCURYTM LNA microRNA Array. The target genes of differentially expressed miRNAs were predicted by bioinformatics software. RESULTS:Twenty-eight miRNAs showed significantly differential expression after H2 O2 treatment, 18 miRNAs upregulated and 10 miRNAs downregulated. The differentially expressed miRNAs may involve in apoptosis of lens epithetium and development of cataract through targeting BCL2L2 and MIP. CONCLUSION: H2 O2 can induce dramatically changes in miRNA profile of HLE, which may play a pivotal role in the pathogenesis and development of cataract.