【摘要】 目的:研究miR-150在心肌梗死后心肌纤维化中的作用及机制。方法:在心梗大鼠,通过过表达miR-150的慢病毒上调miR-150,用RT-PCR及Western bloting检测梗死边缘区col1ɑ1与α-SMA的表达及Masson染色检测纤维化,研究miR-150在心梗纤维化模型中的作用。分离及培养心肌成纤维细胞,用荧光素酶报告载体、慢病毒及c-Myb的si RNA验证miR-150与c-Myb的关系。结果:体内实验表明miR-150在心梗后第2周及第4周心肌表达下调(P<0.001,P<0.05),而上调miR-150明显改善心肌纤维化(P<0.001),抑制col1ɑ1及ɑ-SMA的表达(P<0.01,P<0.05)。体外实验表明c-Myb是miR-150的靶基因且miR-150可通过c-Myb抑制col1ɑ1及ɑ-SMA的表达。结论:miR-150在心肌梗死边缘区表达下降,且可通过靶基因c-Myb改善心肌纤维化。更多还原

【Abstract】 Objective To evaluate the role and mechanism of miR-150 in cardiac fibrosis after MI.Methods A rat model of MI was established by up-regulating miR-150 through overexpressing miR-150 lentivirus.Real-time PCR and Western blot were applied in detecting the expression of collagen1α1 and ɑ-SMA protein ininfarction area border. Masson coloration was applied in measuring fibrosis. Cardiac fibroblasts were isolated andcultured. UTR was used to report the carrier and lentivirus. And c-Myb si RNA was used to verify the relationshipbetween c-Myb and micro RNA-150. Results In vivo,Mi R-150 was down-regulated in myocardium border zonein 14 day and 28 day after infarction(P < 0.001,P < 0.05),and overexpressing miR-150 promoted myocardialfibrosis(P < 0.001),and inhibited the expression of collagen1α1 and ɑ-SMA(P < 0.01,P < 0.05). In vitro,c-Myb was the direct target gene of miR-150,and inhibited the expression of c-Myb resulting in the down-regulation of collagen1α1 and ɑ-SMA,suggesting that the role of miR-150 was achieved by regulating c-Myb.Conclusions Mi R-150 was down-regulated in myocardium border zone,and myocardial fibrosis can be improvedby targeting c-Myb.更多还原