中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (32): 5957-5961.doi: 10.3969/j.issn.1673-8225.2011.32.015

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

神经调节蛋白1诱导小鼠胚胎干细胞向心肌细胞的分化及其机制探讨

王  治，黄  进   

  1. 南京市胸科医院心内科，江苏省南京市  210029
  • 收稿日期:2011-02-14 修回日期:2011-03-14 出版日期:2011-08-06 发布日期:2011-08-06
  • 作者简介:王治☆，男，1977年生，汉族，2009年上海交通大学毕业，博士，主治医师，主要从事冠心病基础与临床研究、干细胞诱导分化等研究。 wangzhi1977nj@hotmail.com
  • 基金资助:

    江苏省自然科基金资助(BK2010119)。

Neuregulin-1 induces embryonic stem cells to differentiate into cardiomyocytes and its mechanism

Wang Zhi, Huang Jin   

  1. Department of Cardiology, Nanjing Chest Hospital, Nanjing  210029, China
  • Received:2011-02-14 Revised:2011-03-14 Online:2011-08-06 Published:2011-08-06
  • About author:Wang Zhi☆, Doctor, Attending physician, Department of Cardiology, Nanjing Chest Hospital, Nanjing 210029, China wangzhi1977nj@hotmail.com
  • Supported by:

    Natural Science Foundation of Jiangsu Province, No. BK2010119*

PDF

385

被引次数

5

摘要：

背景：前期研究发现神经调节蛋白1早期干预能够诱导胚胎干细胞向心肌细胞分化。
目的：进一步观察神经调节蛋白1诱导小鼠胚胎干细胞向心肌细胞分化的途径。
方法：分别观察胚胎干细胞自发与在神经调节蛋白1诱导情况下向心肌细胞的分化，RT-PCR检测胚胎干细胞自发与诱导分化下心肌特异性早期转录因子GATA-4、Nkx2.5 mRNA的表达，细胞免疫组织化学检测胚胎干细胞自发与诱导分化下心肌肌钙蛋白T及磷酸化蛋白激酶B的表达，Western blot半定量分析自发与诱导分化下心肌细胞磷酸化蛋白激酶B的表达。
结果与结论：神经调节蛋白1诱导心肌分化率显著高于自发心肌分化率。神经调节蛋白1呈剂量依赖性上调GATA-4、Nkx2.5 mRNA表达；表皮生长因子样受体拮抗剂及磷脂酰肌醇-3激酶抑制剂阻断了神经调节蛋白1对Nkx2.5 mRNA的表达上调作用。Western blot分析显示神经调节蛋白1诱导组拟胚体中自发性搏动心肌细胞团磷酸化蛋白激酶B的相对表达高于自发心肌分化组。提示神经调节蛋白1可能通过磷脂酰肌醇3激酶/磷酸化蛋白激酶B信号通路诱导胚胎干细胞向心肌细胞分化。

关键词: 神经调节蛋白1, 胚胎干细胞, 心肌细胞分化, 磷脂酰肌醇-3-激酶, 干细胞

Abstract:

BACKGROUND: Previous studies demonstrated that the early invention of neuregulin-1 (NRG-1) can induce embryonic stem cells (ESCs) to differentiate into cardiomyocytes.
OBJECTIVE: To further investigate the pathway of NRG-1 induces embryonic stem cells to differentiate into cardiomyocytes.
METHODS: The process of cardiomyocytes (CMs) differentiated from ESCs with or without NRG-1 treatment was observed continuously. RT-PCR was performed to detect the mRNA expression of early cardiac-restricted transcript factors Nkx2.5 and GATA-4 during differentiation. The protein expression of cardiac troponin T and phosphorylated protein kinase B was determined by immunhistochemistry (IHC), phosphorylated protein kinase B expression of CMs was analyzed by semiquantitative western blot analysis.
RESULTS AND CONCLUSION: The differentiation rate of CMs induced by NRG-1 was significantly higher than spontaneous CMs differentiation rate. NRG-1 exhibited a dose dependent up regulatory effect on the expression of GATA-4, Nkx2.5 mRNA, while NRG-1-induced increase of Nkx2.5 transcripts was inhibited by treatment with receptor antagonist and phosphatidylinositol 3-kinase (PI3K) inhibitor. Western blot analysis confirmed that the expression of phosphorylated protein kinase B of spontaneous beating CMs in embryoid body in NRG-1 induced group was relatively higher than that in spontaneous CMs differentiation group. The results suggested that NRG-1 promotes CMs differentiation of mouse ESCs via PI3K/phosphorylated protein kinase B/ phosphorylated protein kinase B activation.

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