P^(38)MAPK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 P^(38) mitogen-activated protein kinases signaling pathway in the differentiation of bone morphogenetic protein 9 induced C3H10T1/2 stem cell into myocardiocytes

目的:研究P38丝裂原活化蛋白激酶(P38mitogen-activated protein kinases,P38MAPK)通路在骨形态发生蛋白9(Bonemorphogenetic 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。

Objective ：To investigate the role of ps8 mitogen-activated protein kinases （pSsMAPK） signal pathway in the differentiation of bone morphogenetic protein 9（BMP9） induced C3H10T1/2 stem cell into myocardiocytes. Methods：The phospho-p3sMAPK and PSSMAPK were measured by Western blot at 24 h after the C3H10T1/2 stem cell being transfected with pAdEasyBMP9 and the phos- pho-pSSMAPK was detected by immunofluorescence at 24 h after the C3HI 0T1/2 stem cell being transfectcd 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 C3H 10T1/2 stem cell being transfected with pAdEasyBMP9 and treated with SB203580. Results：Transfection with pAdEasyBMP9 produced rapid increase in phospho-pSsMAPK and little change in p38MAPK. Phospho-p38MAPK expressed in nucleus and cytoplasm or in different states of nueleus. SB203580 significantly inhibited the pAdEasyBMP9-induced expression of CX43,cTnT,c^-MHC, GATA4, MEF2C in C3H 10T1/2 stem cell. Conclusions：pssMAPK signal pathway can be activated in C3H10T1/2 stem cell after being transfected with pAdEasyBMP9 and can promote the myocardiocytes differentiation.