糖尿病心肌病小鼠心肌成纤维细胞miR-375表达及其对胶原蛋白Ⅰ合成的影响  

Expression and effect of miR-375 on synthesis of collagen Ⅰ in mice with diabetic cardiomyopathy

作　　者：黄燕[1] 张赢予[1] 王好[1] 周艳芳[1] 张国辉[1] 芮涛[1] 

机构地区：[1]江苏大学附属人民医院心内科,江苏镇江212002

出　　处：《江苏大学学报（医学版）》2013年第3期197-200,206,共5页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(批准号:81170210)

摘　　要：目的:探讨miR-375在糖尿病心肌病心肌纤维化发生发展过程中的作用机制。方法:原代培养小鼠(C57BL/6)心肌成纤维细胞,采用免疫组织化学染色法鉴定细胞纯度。将培养的第2～3代心肌成纤维细胞随机分成对照组和高糖组,高糖组在处理6,12,24 h后,采用实时定量PCR方法检测各组细胞中miR-375的差异性表达。miR-375抑制剂瞬时转染心肌成纤维细胞,实时定量PCR检测转染效率。免疫组织化学染色检测心肌成纤维细胞中Ⅰ型胶原蛋白的表达。结果:原代分离培养的第2代心肌成纤维细胞纯度可达90%;心肌成纤维细胞经高糖处理6,12,24 h后其miR-375的表达有增加趋势;抑制miR-375的表达后,高糖处理的心肌成纤维细胞内Ⅰ型胶原蛋白的表达显著降低。结论:miR-375可能在糖尿病心肌病心肌纤维化中发挥重要的作用。Objective：To investigate the effect of miR-375 on cardiac fibrosis in diabetic cardiomyopathy mice.Methods： Cardiac fibroblasts isolated from hearts of C57BL/6 mice were cultured in DMEM medium（low glucose concentration）.The purity of cardiac fibroblasts were evaluated by immunohistochemical staining.The second and third generation cardiac fibroblasts were randomly divided into control group（5 mmol/L glucose）and high glucose group（25 mmol/L glucose）.The cardiac fibroblasts of high glucose groups were treated by high glucose for different times（6,12,24 h）.The different expression of miR-375 in cardiac fibroblasts was detected by real-time quantitative PCR.miR-375 inhibitor was transiently transfected into cardiac fibroblasts by lipofectamineTM 2000 reagent;transfection efficiency was detected through real-time quantitative PCR.The expression of collagen Ⅰin cardiac fibroblasts was evaluated by immunohistochemical staining.Results： The purity of the second generation of cardiac fibroblasts was above 90%.During the time point 6 to 24 h cardiac fibroblasts treated by high glucose,the expression of miR-375 showed an increase tendency;in high glucose and miR-375 inhibitor treated group,the expression of collagen Ⅰdecreased significantly.Conclusion： MiR-375 might play an important role in diabetic myocardial fibrosis.

关 键 词：MiR-375 糖尿病心肌病 心肌纤维化 

分 类 号：R542.23[医药卫生—心血管疾病]