Integrin β1在维生素C诱导小鼠诱导性多能干细胞向心肌样细胞分化中的作用
《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]
- 期数:
- 2013年第2期
- 页码:
- 151-157167
- 栏目:
- 基础研究
- 出版日期:
- 2013-04-25
文章信息/Info
- Title:
- Effect of integrin β1 on Vc-induced cardiomyocyte differentiation of mouse induced pluripotent stem cells
- Author(s):
- WEI Ting; ZENG Di; OU Dong-bo; DING Lu; LI Xue; ZHENG Qiang-sun
- (Department of Cardiology, Tangdu Hospital, Fourth Military Medical University, Xi’an 710038, Shaanxi, China)
- 分类号:
- R329.21
- DOI:
- -
- 文献标识码:
- A
- 摘要:
- 目的:探讨整联蛋白β1（Integrin β1）在维生素C（Vc）诱导小鼠诱导性多能干细胞（induced pluripotent stem cells，iPSCs）向心肌样细胞(cardiomyocytes，CMs)分化中的作用。方法： 在立式显微镜下，分别摘取胚胎(12.5 d、14.5 d及16.5 d)BALB/c胎鼠、新生（出生1 d，P1）和成年BALB/c小鼠的心脏，提取总RNA，半定量及实时定量PCR分析心脏发育不同时期Integrin β1的表达。利用悬滴培养形成拟胚体(embryoid body，EB)的方法体外诱导iPSCs向心肌样细胞分化，诱导过程中分别添加Vc和integrin β1抑制剂（HMβ1-1）处理，共分为3组：对照组、Vc处理组和Vc+HMβ1-1处理组。诱导分化的第3、5、7天，半定量及实时定量PCR检测Oct4、integrin β1和心肌特异性因子（α-MHC、MLC2a）。用免疫荧光染色法检测心肌特异性结构蛋白心肌肌钙蛋白 I（cTnI）的表达。结果： Integrin β1在心脏胚胎发育时期（E12.5、E14.5、E16.5、P1）的表达递增（P<0.01），成年期表达有所回落（P<0.01）。半定量及实时定量PCR的结果提示，Vc组α-MHC、MLC2a的表达显著高于对照组（P<0.01），加integrin β1抑制剂HM β1-1后表达量降低(P<0.01)。跳动EBs计数的结果提示，诱导分化第10、14、18、22、26天，Vc组跳动EBs的百分率显著高于对照组（P<0.01）；而Vc+ HMβ1-1组跳动EBs的百分率相对于Vc组显著降低（P<0.01）。免疫荧光染色显示，Vc组有较强的cTnI表达，添加integrin β1抑制后，Vc+HMβ1-1组cTnI的表达明显减弱。结论： Vc可促进iPSCs向心肌细胞分化，其机制可能是通过integrin β1介导。
- Abstract:
- AIM:To explore the effect of integrin β1 on Vc-induced cardiomyocyte (CM) differentiation of mouse induced pluripotent stem cells (iPSCs). METHODS: Hearts were acquired, respectively, from embryonic BALB/c fetal mice (12.5 days, 14.5 days, 16.5 days), newborn mice (birth 1 day, P1) and adult BALB/c mice under stereomicroscopy, and total RNA was extracted and then analyzed by semiquantitative and real-time quantitative PCR to observe the expressions of integrin β1 during different periods of heart development. Embryoid bodies (EBs) formed from iPSCs via the hanging drop method in vitro were cultured in three groups: control group (without addition), Vc group, and Vc combined with HM β1-1 (inhibition of integrin β1) group. On the third, fifth and seventh day of differentiation, Oct4, integrin β1 and cardiac-specific factors (α-MHC, MLC2a) were evaluated by semiquantitative RT-PCR and quantitative real-time PCR and the expression of cTnI was evaluated by immunocytochemistry. RESULTS: Integrin β1 expressed increasingly during the embryonic development period of the heart (E12.5, E14.5, E16.5, P1) (P<0.01), but the expression of integrin β1 decreased during the adult stage (P<0.01). Semiquantitative RT-PCR and quantitative RT-PCR, the number of beating EB, and immunocytochemistry staining all suggested that the differentiation efficiency in the Vc group was much higher than in control group (P<0.01). Inhibition of integrin β1 (HMβ1-1) decreased this effect (P<0.01). CONCLUSION: Vc improves the differentiation efficiency of iPSC-derived CMs (iPSCMs). Integrin β1 plays an important role in this process.
参考文献/References
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备注/Memo
- 备注/Memo:
- 收稿日期：2012-11-29.基金项目：国家自然科学基金项目资助（31271039） 通讯作者：郑强荪，主任医师，主要从事心脏组织工程及复杂心律失常射频消融治疗Email:qiangsunzheng@gmail.com 作者简介：魏婷，硕士生 Email:waaiting@126.com
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