糖脂毒性作用下小鼠胰岛miRNA的表达谱变化  

Effect of Glycolipid Toxicity on miRNA Expression Profile in Murine Pancreatic Islets

作　　者：吴倜珺 许茹凤 韩晓[1] 严虹[2] WU Tijun XU Rufeng HAN Xiao YAN Hong(State Key Laboratory of Human Functional Genomics of Jiangsu Province, Nanjing Medical University, Nanjing, 210029, China Department of Clinical Laboratory, Nanjing Chest Hospital, Nanjing, 210029, China)

机构地区：[1] 南京医科大学江苏省人类功能基因组学重点实验室,南京市210029 [2] 南京市胸科医院检验科,南京市210029

出　　处：《医学分子生物学杂志》2016年第6期329-335,共7页Journal of Medical Molecular Biology

基　　金：南京市卫生局医学科技发展基金（No.ZKX13042,QRX11229）

摘　　要：目的：初步探索长期高糖和／或高脂培养对小鼠原代胰岛中miRNA表达的影响。方法选取B6小鼠原代胰岛作为研究对象，提取RNA后通过生物芯片高通量筛选法及RT-PCR法检测并统计胰岛中表达水平变化的miRNA种类；生物信息学方法预测变化显著的miRNA的下游靶基因并通过Western印迹法验证预测结果。结果生物芯片与定量PCR结果均显示，高糖和／或高脂培养后原代胰岛中相当一部分miRNA的表达发生了显著改变，其中miR320在高糖、高脂和高糖高脂联合培养下表达均呈上升趋势，通过生物信息学方法预测p85α为其下游靶基因，干扰小鼠胰岛β细胞系中miR320表达后p85α表达明显下降。结论在本实验中，高糖和／或高脂培养会引起原代胰岛中miR？320的表达水平升高，提示miR？320可能通过参与免疫系统的调控从而对2型糖尿病的发生发展产生作用。Objective To explore the effect of long-term high-glucose and/or high-fat culture on the miRNAs expression in mouse primary pancreatic islets. Methods RNA was extracted from the primary pancreatic islets obtained from B6 mice. MiRNAs that experienced great changes after high-glucose and/or high-fat culture were detected by high-throughput biochip microarray and RT-PCR. The bioinformatics method was used to predict the downstream target genes of miRNAs and the results were verified by western blotting. Results A variety of miRNAs changed significantly in the primary islets after high-glucose and/or high-fat culture, and miR-320 was found to increase under the culture of high glucose and high fat. p85α was predicted to be a downstream target gene of miR320 by bioinformatics method. Western blotting showed that p85α was significantly decreased in pancreatic isletβcells after miR-320 was inhibited. Conclusion The expression of miR-320 is sig-nificantly increased in primary pancreatic islets under high-glucose and/or high-fat culture, sugges-ting that miR-320 may be involved in the development of type 2 diabetes by regulating the immune system.

关 键 词：糖脂毒性 原代胰岛 MIRNA 

分 类 号：R342.3[医药卫生—基础医学]