补肾益气方调控Bcl-2和XIAP蛋白对早孕期人滋养层细胞凋亡的拮抗作用  

Antagonism of Bushen Yiqi Prescription on apoptosis of human first trimester trophoblastic cell by regulating Bcl-2 and XIAP in vitro

作　　者：王海燕[1] 李雪莲[1] 归绥琪[1] 

机构地区：[1]复旦大学附属妇产科医院,上海200011

出　　处：《中华中医药杂志》2009年第6期735-739,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金资助项目(No.C03050402);上海市科学技术发展基金资助项目(No.D2DZ19115);复旦大学985工程资助项目

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。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-G1 and G2/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.

关 键 词：补肾益气方 含药血清 细胞滋养层细胞 增殖 实验研究 

分 类 号：R285[医药卫生—中药学]