小半夏加茯苓汤含药血清对小鼠胚胎干细胞体外增殖及分化的影响

Effect of Xiaobanxia added Fuling decoction-containing serum on the proliferation and differentiation of mouse embryonic stem cells in vitro

[目的]研究小半夏加茯苓汤含药血清对体外培养的小鼠胚胎干细胞(ES)增殖及分化的影响。[方法]噻唑蓝比色法(MTT)检测小半夏加茯苓汤含药血清作用下24、48、72、96 h后的ES细胞活性。半定量逆转录-聚合酶链反应法(RT-PCR)检测小半夏加茯苓汤含药血清作用下ES分化为心肌细胞的能力,判断受试物的胚胎发育毒性。[结果]在促细胞增殖方面:小半夏加茯苓汤含药血清作用下的ES细胞,在连续培养24、48、72、96 h时,其活性水平均呈显著上升趋势,与其他组比较,差异均无统计学意义(P>0.05)。在促细胞分化方面:小半夏加茯苓汤含药血清对ES分化心肌细胞的肌球蛋白重链基因β-MHC的表达能力,与其他组比较,差异均无统计学意义(P>0.05)。[结论]小半夏加茯苓汤含药血清能够促进ES细胞增殖,并能促进ES细胞向心肌细胞分化,提示在该实验剂量下,小半夏加茯苓汤无明显胚胎毒性。

[Objective] To investigate the effect of Xiaobanxia added Fuling decoction-containing serum on the proliferation and differentiation of mouse embryonic stem cells （ESCs） in vitro. [Methods] Isolated ESCs were cultured in vitro. The cells were treated with drug-containing serum and Qingbanxia respectively, the blank serum and fetal bovine serum were used as the control. The proliferation of cells was detected by MTT assay at 24, 48, 72 and 96 h. The differentiation of cells to cardiac myoblasts specific genes （13-MHC） was detected by RT-PCR. [Results] Mouse ESCs were proliferated after treating with drug-containing serum at 24, 48, 72 and 96 h. Differentiation of mouse ESCs were good when treated with this serum. Compared with control group there were no significant differences. （P〉0.05）. [Conclusion] Xiaobanxia added Fuling decoction-containing serum can promote ESCs proliferation, and effectively stimulate ESCs differentiation into cardiac myoblasts. It is possible that Xiaobanxia added Fuling decoction may not have any embryotoxicity.