柴胡三参胶囊含药血清对骨髓间充质干细胞分化为心肌细胞过程中PI3-K/Akt信号通路干预

Intervention of Chaihu Sanshen Capsule Containing Serum on the Differentiation of BMSCs into Myocardial Cell with PI3-K/Akt Signal Pathway

目的：研究柴胡三参胶囊含药血清对骨髓间充质干细胞分化为心肌细胞过程中PI3-K/Akt信号通路的影响,探讨其抗心律失常机制与PI3-K/Akt信号转导通路的相关性。方法：将P3代骨髓间充质干细胞随机分为4组,分别为BMP-2组、BMP-2＋含药血清组、含药血清组及空白血清组,以MTT法测BMSCs增殖情况,确定最佳浓度及最佳时间;加入相应浓度的诱导剂及血清,诱导至最佳时间,Western blotting法检测各组细胞内Akt蛋白表达水平;RT-PCR法检测各组细胞内GATA-4、β-MHC m RNA表达情况。结果：与空白血清组相比,BMP-2组、BMP-2＋含药血清组、含药血清组Akt、GATA-4、β-MHC表达水平均明显升高,统计学具有明显差异（P〈0.01）;与BMP-2组相比,BMP-2＋含药血清组表达水平升高（P〈0.05）,与含药血清组相比,BMP-2＋含药血清组表达水平升高（P〈0.05）;BMP-2组与含药血清组比较,差异无统计学意义（P〉0.05）。结论：柴胡三参胶囊能够通过作用于PI3-K/Akt信号通路诱导骨髓间充质干细胞向心肌细胞分化。

Objective：To study the effects of Chaihu Sanshen Capsule（CHSSC）containing serum on the differentiation of BMSCs into myocardial cell with PI3-K/Akt signal pathway and explore the mechanism of anti arrhythmia effect of CHSSC. Methods：P3 BMSCs were randomly divided into four groups,its group BMP-2,group BMP-2 with containing serum,containing serum group and blank serum group. MTT assay was used to detect the proliferation of BMSCs,determing the best concentration and the best time,joining the corresponding concentration of inducer and serum,inducing to optimal time,detecting p-Akt levels of each group with WB and the expression of GATA-4 and β-MHC in each group of cells with RT-PCR. Results：The expression of BMP-2 group,BMP-2 with containing serum group,containing serum group＇s p-Akt,GATA-4 and β-MHC levels were higher than the blank serum group,the difference was statistically significant（P〈0.01）,the expression level of BMP-2 with containing serum was higher than that of BMP-2 group（P〈0.05）,containing serum group（P〈0.05）,the difference of BMP-2 group and containing serum group was not statistically significant（P〈0.05）. Conclusion：CHSSC can induce the differentiation of bone marrow mesenchymal stem cells into cardiomyocytes by acting on PI3-K/Akt signaling pathway.