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血管紧张素Ⅱ诱导人骨髓间充质干细胞分化为心肌样细胞(PDF)

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2006年第3期
页码:
258-261
栏目:
基础研究
出版日期:
2006-06-25

文章信息/Info

Title:
AngiotensinⅡ induces cardiomyocytes from human bone marrow-derived mesenchymal stem cells
作者:
苑媛; 吕安林; 陈丹; 刁繁荣; 李军杰; 胡小菁; 易甫
第四军医大学西京医院心血管内科， 陕西 西安 710032
Author(s):
YUAN Yuan;  Lv An-lin;  CHEN Dan;  DIAO Fan-rong;  LI Jun-jie;  HU Xiao-jing;  YI Fu
Department of Cardiology, Xijing Hospital, Fourth Military Medical University, Xi′an,Shaanxi 710032, China
关键词:
骨髓间充质干细胞; 心肌样细胞; 血管紧张素Ⅱ
Keywords:
bone marrow-derived mesenchymal stem cells; cardiomyocyte; angiotensinⅡ
分类号:
R331.31
DOI:
-
文献标识码:
A
摘要:
目的 研究血管紧张素Ⅱ(AngⅡ)作用于人骨髓间充质干细胞(hMSCs)后，心肌特异蛋白及细胞外信号调节激酶（ERK 1／2）的表达变化。方法 第8代hMSCs用0.1 μmol/L AngⅡ孵育24 h。倒置显微镜下观察细胞形态变化,免疫细胞化学法标测心肌肌钙蛋白I(cTnI)、间隙连接蛋白43（connexin 43）及α肌动蛋白(αactin)表达情况,反转录PCR法检测细胞cTnI表达，透射电镜观察分化的心肌样细胞内是否有肌丝存在，Western blot法检测细胞ERK 1／2总蛋白表达,流式细胞仪测定心肌样细胞分化率。结果 与对照组比较，AngⅡ组hMSCs诱导后细胞形态改变，第2周开始表达αactin及connexin 43，第3周开始表达cTnI，AngⅡ组细胞ERK 1／2的蛋白表达水平逐渐增加，流式细胞仪测得心肌样细胞分化率为(23±3)％。结论 AngⅡ在体外可能经ERK通路诱导hMSC向心肌样细胞分化。
Abstract:
AIM To investigate the changes of specific protein of myocardium and extracellular signal regulated kinase 1／2(ERK1／2) of human bone marrowderived mesenchymal stem cells (hMSCs) induced to cardiomyocytes in vitro with the treatment of angiotensinⅡ (AngⅡ). METHODS 4 ml of bone marrow was obtained from normal adults and hMSCs were isolated by proportion 1.077 Ficoll and were cultured in LGDMEM for more than 8 passage. hMSCs were treated with 0.1 μmol/L AngⅡ for 24 hours. The differentiation of MSCs was observed with a phasecontrast mictmcope. The induced cells were evaluated by immunocytochemistry staining and RTPCR analysis, and the myofilament structure was observed by electron microscopy observed. The ERKI／2 at different time points was measured by Western blot and cardiomyocytes differentiation was examined by flow cytometer. RESULTS After the induction by AngⅡ, some hMSCs became bigger and longer. The cells were arrayed in a similar direction. The cells induced by AngⅡ were stained positively for aactin, connexin 43 and cTnI. RTPCR showed that these cells expressed Cardiac Troponin I. The expression of ERK1／2 increased in AngⅡ group. CONCLUSION AngⅡ induces human bone marrowderived mesenchymal stem cells into cardiomyocytes and ERK1／2 MAPK pathway may be involved in cardiomyocytes differentiation of hMSCs cultures.

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备注/Memo

备注/Memo:
收稿日期：2006-01-06.基金项目：西京医院临床高新技术项目资助(No. XJGX0510M08) 通讯作者：吕安林，副教授，主要从事冠心病诊断与治疗研究 Tel：（029）84775183 Email:lvanlin@fmmu.edu.cn 作者简介：苑媛，硕士 Tel：(029)84773464 Email:jujube@fmmu.edu.cn
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