【摘要】 目的:探讨补肾益气方对正常早孕期人滋养层细胞凋亡的调节作用及机制。方法:体外培养正常早孕期人细胞滋养层细胞,分别加入空白对照组、5%含药血清组、10%含药血清组、20%含药血清组,用噻唑蓝(MTT)法、流式细胞仪检测含药血清作用24、48、72h后细胞的增殖活性,Caspase-3活性检测试剂盒检测细胞Caspase-3酶活性,Western blot检测Fas、FasL、Bcl-2和XIAP蛋白表达。结果:含药血清培养24、48、72h后,细胞在490nm波长的吸光度值增加(P<0.01);sub-G1期细胞减少(P<0.01),S期细胞增加(P<0.01),G2/M期细胞明显减少(P<0.01),Caspase-3酶活性降低;含药血清对Fas和FasL表达均有上调作用,呈剂量依赖式诱导(P<0.05);Bcl-2蛋白表达水平上升,随着浓度增加,表达量也增加,同时伴随XIAP水平升高(P<0.05)。结论:补肾益气方通过提高凋亡抑制蛋白Bcl-2水平,从而增强XIAP表达,抑制Caspase-3活性,抑制细胞死亡。FasL表达水平的上调有利于母胎免疫耐受的维持。更多还原

【Abstract】 Objective:To investigate the effect and mechanism of Bushen Yiqi Prescription on apoptosis of human first trimester trophoblasts.Methods:Isolated from first trimester placental tissues,human trophoblasts cultured in media with 5%, 10%,20%drug-containing serum for a 24,48 or 72h period.Then we measured cell viability by MTT assay,cell DNA content by flow cytomester,cell Caspase-3 enzyme activity by Caspase-3 active assay kit,and,cell Fas,FasL,Bcl-2 and XIAP protein.Results:Absorbency of trophoblast in 490nm wavelength increased greatly after being incubated with drug-containing serum for 24,48,and 72 h(P<0.01).Cells cultured with drug-containing serum in S phase were more and by contrast,in sub-G 1 and G 2 /M phase were less than those of control group.In addition,Caspase3 enzyme activity decreased.Also,the expression of Fas,FasL, and Bcl-2 showed dose dependent with the concentration of drug-containg serum,and it was followed with the increase of XIAP level.Conclusion:Bushen Yiqi Prescription antagonized the apoptosis of human first trimester trophoblasts by increasing the level of Bcl-2 to promote XIAP expression.Increasing of FasL expression facilitated the immunological tolerance of maternal fetal interface.更多还原