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Expression and effect of miR375 on synthesis of collagen Ⅰ in mice with diabetic cardiomyopathy |
HUANG Yan, ZHANG Ying-yu, WANG Hao, ZHOU Yan-fang, ZHANG Guo-hui, RUI Tao |
Department of Cardiology, the Affiliated People′s Hospital of Jiangsu University, Zhenjiang Jiangsu 212002, China |
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Abstract Objective:To investigate the effect of miR375 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 miR375 in cardiac fibroblasts was detected by realtime quantitative PCR.miR375 inhibitor was transiently transfected into cardiac fibroblasts by lipofectamineTM 2000 reagent; transfection efficiency was detected through realtime 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 miR375 showed an increase tendency;in high glucose and miR375 inhibitor treated group,the expression of collagen Ⅰdecreased significantly. Conclusion: MiR375 might play an important role in diabetic myocardial fibrosis.
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Received: 20 March 2013
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