过表达miR-130a-3p对心肌细胞肥大的缓解作用研究  

Overexpression of miR-130a-3p attenuates cardiomyocyte hypertrophy

作　　者：王晓姣 曲径 李东旭 李君丽[1,3] 吴文超[1] 刘小菁[1,3] WANG Xiaojiao;QU Jing;LI Dongxu;LI Junli;WU Wenchao;LIU Xiaojing(Lab of Cardiovascular Diseases,Regenerative Medicine Research Center,West China Hospital,Sichuan University,Chengdu 610041,P.R.China;Department of Cardiac Surgery,West China Hospital,Sichuan University,Chengdu 610041,P.R.China;Department of Cardiology,West China Hospital,Sichuan University,Chengdu 610041,P.R.China)

机构地区：[1]四川大学华西医院,再生医学研究中心,心血管疾病研究室,成都610041 [2]四川大学华西医院心脏外科,成都610041 [3]四川大学华西医院心内科,成都610041

出　　处：《生物医学工程学杂志》2020年第2期340-348,共9页Journal of Biomedical Engineering

基　　金：国家自然科学基金项目(11672197)。

摘　　要：本研究旨在探索miR-130a-3p对心肌细胞肥大的作用及其可能机制。通过胸主动脉缩窄法(TAC)构建压力超负荷所致心肌肥厚小鼠模型。使用去甲肾上腺素(NE)刺激SD乳鼠原代心肌细胞(NRCMs)及H9c2大鼠心肌细胞系,诱导这两种心肌细胞发生肥大表型转变。检测miR-130a-3p的表达变化,并进一步探索其对心肌细胞肥大是否有调控作用。结果表明,miR-130a-3p在肥厚心肌组织、肥大NRCMs及H9c2细胞中的表达均明显降低。给予miR-130a-3p mimics使其过表达后,H9c2细胞中肥大标志基因心房利钠肽(ANP)、脑利钠肽(BNP)和肌球蛋白重链β(β-MHC)的表达较对照组(mimics N.C.+NE组)明显下调,且细胞面积明显减小。而给予miR-130a-3p inhibitor抑制其表达后,肥大心肌细胞中ANP、BNP、β-MHC的表达进一步上升,且细胞面积进一步增加。Western blot检测发现,过表达miR-130a-3p后心肌细胞中磷酸化Akt和磷酸化mTOR的表达水平下调。以上结果提示,miR-130a-3p mimics可缓解心肌细胞肥大的程度;其inhibitor则可使心肌细胞肥大进一步加剧。过表达miR-130a-3p可能通过影响Akt通路来缓解H9c2心肌细胞肥大的程度。This study aimed to explore the role of miR-130a-3p in cardiomyocyte hypertrophy and its underlying mechanisms. Pressure-overload induced myocardial hypertrophy mice model was constructed by thoracic aortic constriction(TAC). In vitro, norepinephrine(NE) was used to stimulate neonatal rat cardiomyocytes(NRCMs) and H9c2 rat cardiomyocytes to induce hypertrophic phenotypes. The expression of miR-130a-3p was detected in mice hypertrophic myocardium, hypertrophic NRCMs and H9c2 cells. The mimics and inhibitors of miR-130a-3p were transfected into H9c2 cells to observe the role of miR-130a-3p on the hypertrophic phenotype change of cardiomyocytes separately.Furthermore, whether miR-130a-3p regulated hypertrophic related signaling pathways was explored. The results showed that the expression of miR-130a-3p was significantly decreased in hypertrophic myocardium, hypertrophic NRCMs and H9c2 cells. After transfection of miR-130a-3p mimics, the expression of hypertrophic marker genes, atrial natriuretic peptide(ANP), brain natriuretic peptide(BNP) and β-myosin heavy chain(β-MHC), and the cell surface area were notably down-regulated compared with the control group(mimics N.C. + NE group). But after transfection of miR-130a-3p inhibitor, the expression of ANP, BNP and β-MHC in H9c2 cells increased significantly, and the cell area increased further. By Western blot, it was found that the protein phosphorylation level of Akt and mTOR were down-regulated after over-expression of miR-130a-3p. These results suggest that miR-130a-3p mimics may alleviate the degree of cardiomyocyte hypertrophy, meanwhile its inhibitor can further aggravate cardiomyocyte hypertrophy. Over-expression of miR-130a-3p may attenuate cardiomyocytes hypertrophy by affecting the Akt pathway.

关 键 词：miR-130a-3p 心肌肥厚 心肌细胞肥大 压力超负荷 AKT 

分 类 号：R54[医药卫生—心血管疾病]