摘要
目的在转化生长因子β1(transforming growth factorβ1,TGFβ1)诱导的分化模型中探讨RhoA信号对于磷酸鞘氨醇受体1/3(sphingosine 1-phosphate receptor 1/3,S1PR1/3)介导骨髓间充质干细胞(bone marrow mesenchymal stem cell,BMSC)向肌成纤维细胞(myofibroblast,MF)分化的影响及其机制。方法分离并培养小鼠原代BMSC,TGFβ1诱导其分化。采用qRT-PCR检测分化标志物平滑肌肌动蛋白α(α-smooth muscle actin,αSMA)、Ⅰ型胶原[procollagenα1(Ⅰ),Colα1(Ⅰ)]和Ⅲ型胶原[Colα1(Ⅲ)]的mRNA表达;采用Pull-down试剂盒检测RhoA的活化。结果 TGFβ1能够显著上调BMSC细胞中分化标志物αSMA、Colα1(Ⅰ)和Colα1(Ⅲ)的mRNA表达,呈现剂量依赖效应。TGFβ1能够激活小G蛋白RhoA,该作用可以被S1PR1或S1PR3拮抗剂所阻断。RhoA抑制剂C3转移酶能够阻断TGFβ1诱导的αSMA、Colα1(Ⅰ)和Colα1(Ⅲ)mRNA水平的上调。结论RhoA信号参与了S1PR1/3介导的BMSC向MF的分化。
Objective Aim of this study is to explore the role of RhoA in transforming growth factor β1 （TGFβ1）-induced bone marrow mesenchymal stem cell （BMSC） differentiation to myofibroblast （MF） mediated by sphingosine 1-phosphate receptor 1/3 （S1PR1/3）.Methods Serum-starved primary cultured mouse BMSCs were stimulated by a series dose of TGFβ1.Expression of α-smooth muscle actin （αSMA）,procollagen α1（Ⅰ） [Col α1（Ⅰ）] and procollagen α1（Ⅲ） [Col α1（Ⅲ）] was measured by qRT-PCR.Activated RhoA induced by TGFβ1 in BMSCs was determined by Pull-down assay.Results TGFβ1 induced an up-regulation of αSMA,Col α1（Ⅰ） and Col α1（Ⅲ） mRNA levels in BMSCs.TGFβ1 triggered the activation of RhoA,which can be blocked by S1PR1/3 antagonists.RhoA inhibitor C3 Transferase reversed the up-regulation of αSMA,Col α1（Ⅰ） and Col α1（Ⅲ） mRNA expression induced by TGFβ1.Conclusion RhoA is involved in the differentiation of BMSC to MF induced by TGFβ1.
作者
杨苑儒
周璇
张若辰
李丽英
杨乐
Yang Yuanru Zhou Xuan Zhang Ruochen Li Liying Yang Le(Grade 2013 Medical Laboratory Animal Science, Capital Medical University, Beijing 100069 Department of Cell Biology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China)
出处
《首都医科大学学报》
CAS
北大核心
2017年第2期277-281,共5页
Journal of Capital Medical University
基金
北京市自然科学基金(7164237)
北京市优秀人才培养资助(2014000020124G156)~~
关键词
小鼠
骨髓间充质干细胞
磷酸鞘氨醇受体
RHOA
mouse
bone marrow mesenchymal stem cell
sphingosine 1-phosphate receptor
RhoA
