P38MAPK通路在BMP9诱导C3H10T1/2干细胞向心肌样细胞分化中的作用

Role of P38 mitogen-activated protein kinases signaling pathway in the differentiation of bone morphogenetic protein 9 induced C3H10T1/2 stem cell into myocardiocytes

摘要:

目的：研究P38丝裂原活化蛋白激酶（P38 mitogen-activated protein kinases，P38MAPK）通路在骨形态发生蛋白9（Bone morphogenetic protein 9，BMP9）诱导 C3H10T1/2干细胞向心肌样细胞分化中的作用。方法：免疫印迹检测经pAdEasyBMP9诱导24 h的C3H10T1/2干细胞磷酸化P38MAPK和P38MAPK表达水平，免疫荧光定位磷酸化P38MAPK；SB203580抑制C3H10T1/2干细胞磷酸化P38MAPK活性，pAdEasyBMP9诱导21 d时免疫印迹检测心肌特异性蛋白CX43、cTnT表达，免疫荧光检测心肌特异性蛋白cTnT、α-MHC表达，RT-qPCR检测心肌特异性基因GATA4、MEF2C表达。结果：pAdEasyBMP9促进磷酸化P38MAPK表达增高，却不影响P38MAPK表达水平；磷酸化P38MAPK有表达于胞浆和胞核、仅表达于胞核的不同状态。SB203580可显著性地抑制由pAdEasyBMP9诱导的C3H10T1/2干细胞CX43、cTnT、α-MHC、GATA4、MEF2C表达。结论：pAdEasyBMP9诱导C3H10T1/2干细胞能激活P38MAPK通路，并促进其向心肌样细胞分化。

Abstract:

Objective：To investigate the role of P38 mitogen-activated protein kinases（P38MAPK） signal pathway in the differentiation of bone morphogenetic protein 9（BMP9） induced C3H10T1/2 stem cell into myocardiocytes. Methods：The phospho-P38MAPK and P38MAPK were measured by Western blot at 24 h after the C3H10T1/2 stem cell being transfected with pAdEasyBMP9 and the phos－pho-P38MAPK was detected by immunofluorescence at 24 h after the C3H10T1/2 stem cell being transfected with pAdEasyBMP9. The cardiac-specific proteins cTnT and CX43 were measured by Western blot，the cardiac-specific proteins cTnT and α-MHC were measured by immunofluorescence，the cardiac-specific gene GATA4 and MEF2C expressions were detected by RT-qPCR at 21 d after the C3H10T1/2 stem cell being transfected with pAdEasyBMP9 and treated with SB203580. Results：Transfection with pAdEasyBMP9 produced rapid increase in phospho-P38MAPK and little change in P38MAPK. Phospho-P38MAPK expressed in nucleus and cytoplasm or in different states of nucleus. SB203580 significantly inhibited the pAdEasyBMP9-induced expression of CX43，cTnT，α-MHC，GATA4，MEF2C in C3H10T1/2 stem cell. Conclusions：P38MAPK signal pathway can be activated in C3H10T1/2 stem cell after being transfected with pAdEasyBMP9 and can promote the myocardiocytes differentiation.