p38对骨髓间充质干细胞成肌分化作用的研究

THE ROLE OF p38 ON THE DIFFERENTIATION OF MSCs TO MYOBLASTS

目的以5氮杂胞苷(5AzaCR)诱导骨髓间充质干细胞(MSCs)成肌分化,探讨生肌调控因子中Myf5的表达及信号转导通路p38在此分化过程中的作用。方法分离、纯化MSCs,以10μmol/L5AzaCR诱导其向成肌细胞分化,RTPCR测定Myf5的表达,免疫组化检测肌球蛋白(myosin)的表达,Westernblot检测p38信号通路特异抑制剂SB203580作用前后磷酸化p38的变化。结果SB203580作用后,Myf5表达由6h延迟至9h,诱导12d部分MSCs表达myosin,18d表达的数量及程度明显增加;5AzaCR诱导后,磷酸化p38蛋白水平较作用前增强,但在SB203580抑制后明显受抑。结论5AzaCR诱导后,MSCs表达生肌调控因子并向成肌细胞分化,p38在此分化过程中起着正向信号转导作用。

Aim: Inducing mesenchymal stem cells (MSCs) differentiate to myoblasts with 5 azacytidine(5 Aza CR),investigating the expression of Myf5 and the role of the signal transduction case of p38 in all the course of differentiation. Methods: Seperating and purifying bone marrow derived MSCs, inducing MSCs differentiation to myoblasts with 10 μmol/L 5 Aza CR, assaying the gene expre ssion time of Myf5 with RT PCR method, the antigen expression of myosion with immunohistochemistray method and observing the changes of the activity of phosphorylation p38 before and after inhibited by SB203580 with Western blot method. Results: MSCs begin to express Myf5 delaied to the 9th day after inhibited by SB203580. Some of MSCs express myosion at the 7th day after induced; The phosphorylation p38 activity of MSCs enhanced after induced by 5 Aza CR but obviously decreased after inhibited by SB203580.Conclusion: MSCs can express myogenic regulator factors and orientation differentiate to myoblasts after induced by 5 Aza CR, p38 really have a positive signal transduction effection in this course.