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环巴胺(CPA)对牛脂肪间充质干细胞周期及分化的影响
黄亚斐1,刘扬2,杨春荣3,魏著英3,李东芳3,胡思敏3,白春玲1
1. 内蒙古大学
2. 西北农林科技大学动物科技学院
3. 内蒙古大学哺乳动物生殖生物学及生物技术教育部重点实验室
Effects of Cyclopamine Treatment on Cell Cycle and Differenciation of Bovine(Bos taurus) Adipose Mesenchymal Stem Cells
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摘要 间充质干细胞(mesenchymal stem cell, MSC)来源于中胚层的多潜能干细胞,具有分化为成骨细胞、软骨细胞和脂肪细胞等终末分化细胞的能力。本实验研究了hedgehog信号通路的特异性抑制剂环巴胺(cyclopamine, CPA)对牛(Bos taurus)脂肪间充质干细胞(adipose mesenchymal stem cells, AMSCs)的细胞分裂周期及成骨与成脂诱导分化的影响。首先获得牛AMSCs,然后用CPA处理AMSCs,用流式细胞仪分析和Real-time PCR方法检测CPA对细胞周期及定向分化过程中关键基因表达的影响。结果显示,CPA以剂量依赖的方式影响AMSCs的细胞周期进程,并且引起细胞停滞在G0/G1期;后经成脂和成骨诱导后,细胞形态没有显著变化;但是用Real-time PCR的方法检测诱导分化过程中成脂分化关键基因过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptor γ, PPARγ)与成骨分化关键基因成骨细胞转录因子2(runt-related transcription factor 2, Runx2)的表达量显示,CPA对两种分化相关基因的表达量都有显著影响:在成脂分化过程中,4 d后处理组的PPARγ比对照组的表达量显著提高(P<0.01);在成骨分化过程中,处理组Runx的表达量从开始一直显著低于对照组(P<0.01)。结果表明,CPA处理对成脂与成骨没有形态学方面的影响,但是能够引起关键基因表达量的显著变化。研究结果为研究hedgehog信号通路在间充质干细胞的增殖与分化过程中的作用提供参考依据。
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作者相关文章
黄亚斐
刘扬
杨春荣
魏著英
李东芳
胡思敏
白春玲
关键词 脂肪间充质干细胞Hedgehog信号通路环巴胺细胞周期分化    
Abstract:Mesenchymal stem cells (MSCs) are multipotent stem cells derived from the mesoderm, which have ability to differentiate into osteoblasts, chondrocytes and adipocytes. In this experiment, the effects of cyclopamine (CPA), a specific inhibitor of hedgehog signaling pathway, on cell division cycle and adipogenic induction and osteogenic induction of bovine(Bos taurus) adipose mesenchymal stem cells (AMSCs) were studied. After isolation and identification of AMSCs, CPA was used to treat AMSCs, then effects of CPA on cell cycle and adipocyte differentiation and osteoblast differentiation associated gene expressions were investigated using flow cytometry and Real-time PCR. The results indicated that treatment of AMSCs with CPA at different concentrations affected cell cycle progression in a dose-dependent manner. CPA resulted in cells arrested at G0/G1 stages. After adipogenic induction and osteogenic induction, no obvious morphological differences was observed in the formation of adipocytes and osteoblasts. However, the Real-time PCR detections of the induced cells showed that both peroxisome proliferator-activated receptorγ(PPARγ, critical adipogenic gene) and runt-related transcription factor 2(Runx, critical osteogenic gene) were significantly changed during adipogenic induction or osteogenic induction. PPARγ expression was significantly increased after 4 d treatment compared to the control (P<0.01), Runx expression was lower in treatment group than the control (P<0.01). These results suggested CPA induced AMSCs arrested at G0/G1 stages and CPA treatments of AMSCs affected adipogenic and osteogenic induction in gene expression level. This experiment provides a basis for the study of hedgehog signaling pathway in AMSCs proliferation and differentiation.
Key wordsAdipose mesenchymal stem cells    Hedgehog signaling pathway    Cyclopamine    Cell cycle    Differentiation
收稿日期: 2013-05-06     
通讯作者: 白春玲   
引用本文:   
黄亚斐1,刘扬2,杨春荣3,魏著英3,李东芳3,胡思敏3,白春玲1. 环巴胺(CPA)对牛脂肪间充质干细胞周期及分化的影响[J]. , 2013, 21(10): 1221-1230.
, , , , , ,. Effects of Cyclopamine Treatment on Cell Cycle and Differenciation of Bovine(Bos taurus) Adipose Mesenchymal Stem Cells. , 2013, 21(10): 1221-1230.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2013/V21/I10/1221
 
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