资冲颗粒大鼠含药血清诱导小鼠胚胎干细胞向生殖细胞分化相关基因的表达

Study on the related gene about mouse embryonic stem cells differentiating into germ cells induced by rat Zichong Granule drug serum

目的:研究资冲颗粒含药血清诱导小鼠胚胎干细胞(m ESC)生殖分化相关基因的表达。方法:以终浓度5%资冲颗粒含药血清干预m ESC,以5%人卵泡液及小鼠卵巢组织共培养作为对照。培养21d后利用实时荧光定量PCR测定干预后细胞内基因Oct4、Stra8、Tp2、Zp3、Itgb1、Gdf9的表达。结果:与m ESC未干预状态相比较,3组干预后生殖分化相关基因表达均有明显变化;同人卵泡液组比较,含药血清组细胞内基因Tp2、Gdf9表达量显著上调(P=0.00、P=0.04);同卵巢组织组比较,含药血清组多数基因上调幅度较大,但两组间无统计学意义。结论:3种诱导方法均促使了m ESC向生殖细胞方向分化;资冲颗粒含药血清或能加速m ESC向生殖细胞方向定向分化的进程。

Objective： To investigate the related gene about mouse embryonic stem cells（m ESC） differentiating into germ cells induced by rat Zichong Granule（ZCG） drug serum.Methods： 5% of rat ZCG drug serum was used to culture the m ESC,while 5% of human follicular fluid and mice ovarian tissue were used to co-culture as the control.After 21d-culture,real-time PCR was used to detect the mRNA expressions of Oct4,Stra8,Tp2,Zp3,Itgb1 and Gdf9 in cultured cells.Results： Compared with the un-intervention state,genes related to reproduction differentiation had obvious changes.Compared with the human follicular fluid group,the mRNA expressions of Tp2 and Gdf9 in drug serum group were both increased（P=0.00,P=0.04）.Compared with the ovarian tissue group,most of the genes in drug serum group were increased,with no statistical significance.Conclusion： All of the three methods above could induce m ESC to differentiate into germ cells,especially in the effect of rat ZCG drug serum.