哌唑嗪(PRA)拮抗去甲肾上腺素(NE)诱导人类胚胎干细胞来源的心肌细胞(hESCs-CMs)肥大  

The effect of prazosin (PRA) on cardiac hypertrophy induced by norepinephrine (NE) in human embryonic stem cells-cardiomyocytes (hESCs-CMs)

作　　者：黄洁 裴晏梓 刘阳 陈海燕 孙宁 HUANG Jie;PEI Yan-zi;LIU Yang;CHEN Hai-yan;SUN Ning(Department of Physiology and Pathophysiology,School of Basic Medical Sciences,Fudan University,Shanghai 200032,China;Department of Echocardiography,Zhongshan Hospital,Fudan University,Shanghai 200032,China)

机构地区：[1]复旦大学基础医学院生理与病理生理学系,上海200032 [2]复旦大学附属中山医院心脏超声诊断科,上海200032

出　　处：《复旦学报（医学版）》2019年第3期309-315,共7页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金(31571527)~~

摘　　要：目的在人类胚胎干细胞来源的心肌细胞(human embryonic stem cells-cardiomyocytes,hESCs-CMs)上,研究α受体拮抗剂-哌唑嗪(prazosin,PRA)对去甲肾上腺素(norepinephrine,NE)诱导的心肌肥大效果的影响。方法通过体外二维培养分化的方法将hESCs-H7细胞系定向诱导分化为CMs,并通过imageJ软件计算免疫荧光染色图片来检测CMs面积的改变,qRT-PCR方法检测CMs肥大相关基因mRNA表达的改变,通过Leica显微镜FelixGX细胞动缘检测系统来测定CM收缩力及跳动情况的改变。结果20 μmol/L NE处理可使hESCs-CMs面积明显增大,肥大相关基因mRNA水平表达上调,收缩力及跳动频率增加;15 μmol/L PRA处理后hESCs-CMs面积相较NE组有所降低,肥大相关基因 BNP、TNNT2 mRNA表达明显降低,CM收缩力及跳动频率明显减弱。结论α受体拮抗剂PRA在hESCs-CMs上通过抑制NE诱导的细胞面积增大、肥大相关基因表达上调、收缩力增强以及跳动频率增加,从而改善NE诱导的心肌肥大。Objective To investigate the effect of prazosin (PRA),α receptor antagonist, on cardiac hypertrophy induced by norepinephrine (NE) in human embryonic stem cells-cardiomyocytes-cardiomyocytes (hESCs-CMs). Methods Human embryonic stem cells (H7) were differentiated into CMs through 2D culture in vitro .Immunofluorescence staining images were calculated by software imageJ to analyze the cell size changes of CMs.The expression level of hypertrophy associated genes in CMs were analyzed by real time PCR.The contractility and of Leica beating rates of CMs were examined by FelixGX detection system. Results NE (20 μmol/L) significantly induced the increase of cell size,the expression of hypertrophy-associated genes,contraction force and beating rates of hESCs-CMs.PRA (15 μmol/L) treatment significantly inhibited NE-induced increase of cell size,mRNA expressions of BNP and TNNT2 , contraction force and beating rates of hESCs-CMs. Conclusions PRA can alleviate NE-induced cardiac hypertrophy in hESCs-CMs by reducing cell size,down-regulatingthe expression of hypertrophy-associated genes,reducing contractility and beating frequency.

关 键 词：哌唑嗪(PRA) 去甲肾上腺素(NE) 胚胎干细胞(ESC) 心肌分化 心肌肥大 

分 类 号：Q786[生物学—分子生物学] R542.2[医药卫生—心血管疾病]