心肌细胞特异性miR-30b转基因小鼠的建立及miR-30b功能的探索

Pub Date : 2014-01-01 DOI:10.3724/SP.J.1206.2013.00206
Yuan-yuan Fan, B. Long, Fang Liu, Luyu Zhou, Kun Wang, Peifeng Li
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引用次数: 1

摘要

MicroRNAs(miRNAs)阵列结果显示,在MYH7错义突变携带者的家族性肥厚性心肌病患者的心脏组织以及小鼠心力衰竭模型中,miR-30b的表达下调,这意味着miR-30b可能在心脏病中发挥重要作用。为了研究miR-30b在体内的功能,我们在α-肌球蛋白重链(α-MHC) 5.5 kb启动子的控制下,构建了过表达miR-30b的转基因小鼠系。qRT-PCR结果显示,miR-30b转基因小鼠心脏组织中miR-30b水平显著升高(P < 0.05)。miR-30b转基因小鼠没有表现出明显的心脏/体重和左心室(LV)/体重变化和心肌结构异常。目前对miR-30b调控心肌梗死的机制知之甚少。采用冠状动脉结扎法构建I/R模型,以假手术小鼠为对照。生化检测结果和TTC-Evans蓝结果显示,与对照组相比,转基因小鼠缺血再灌注后LDH、CK、cTn的释放量较低(P < 0.05),心肌梗死面积较小(P < 0.05)。超声心动图结果显示,转基因小鼠的心功能较对照组有明显改善。综上所述,miR-30b对缺血再灌注损伤具有保护作用。为预防和治疗心肌梗死提供了新的治疗途径。
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Establishment of Cardiomyocyte-specific miR-30b Transgenic Mice and Exploring The Function of miR-30b
MicroRNAs(miRNAs) array results have shown that the expression of miR-30b is downregulated in heart tissues from patients with familial hypertrophic cardiomyopathy who were carriers of missense mutations in the MYH7,and also in a murine heart failure model,implying that miR-30b might play an important role in heart diseases.To study miR-30b in vivo function,we generated a transgenic mouse line overexpressing miR-30b under the control of the 5.5 kb promoter of α-myosin heavy chain(α-MHC).qRT-PCR results demonstrated that miR-30b was significantly increased in the heart tissues of miR-30b transgenic mice(P 0.05).miR-30b transgenic mice did not exhibit significant heart/body weight and left ventricular(LV)/body weight changes and abnormal myocardium structure.At present,little is known about how miR-30b regulates myocardial infarction.We constructed I/R models by the coronary artery ligation method and sham-operated mice were used as controls.Biochemical detection results and TTC-Evans blue results showed that after ischemia-reperfusion,these transgenic mice had lower releases of LDH,CK and cTn玉(P 0.05)and their hearts exhibited a smaller infarct size compared to those from control mice(P 0.05).Echocardiographic results indicated that cardiac function of transgenic mice was markedly improved compared to that of control mice.In conclusion,miR-30b has protective effect upon ischemic-reperfusion injury.And it may provide a new therapeutic approach for preventing and treating myocardial infarction.
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