米非司酮抑制人子宫内膜基质细胞增殖  

Mifepristone inhibits the proliferation of cultured human endometrium stromal cells

作　　者：郭永[1] 赵爱华[2] 苏晨[1,3] 丁宁[1] 张瑶楠[1] 刘庆[1] 王乾兴[3] 

机构地区：[1]国家人口计生委科学技术研究所内分泌室,北京100081 [2]解放军第309医院妇产科,北京100091 [3]遵义医学院细胞生物学与遗传学教研室,遵义563000

出　　处：《生殖医学杂志》2014年第6期475-479,共5页Journal of Reproductive Medicine

基　　金：北京市自然科学基金资助项目(7132160);国家"十二五"科技支撑计划(2012BAI31B06)

摘　　要：目的在改良子宫内膜细胞原代分离培养方法的基础之上,探讨米非司酮对原代分离培养的人子宫内膜基质细胞增殖的抑制作用。方法利用改良后的"二次消化法"分离培养增殖期人子宫内膜基质和上皮细胞,进行细胞形态学观察,并通过流式细胞术检测人子宫内膜基质和上皮细胞的波形蛋白和角蛋白表达阳性的细胞比率,计算分离培养的基质和上皮细胞的纯度;采用四甲基偶氮唑盐(MTT)法检测米非司酮对原代分离培养人子宫内膜基质细胞的半数抑制浓度(IC50)。结果原代分离培养的基质细胞较上皮细胞生长迅速。流式细胞检测结果显示,分离培养的基质细胞波形蛋白阳性率达97%,上皮细胞角蛋白阳性率达90%。米非司酮处理人子宫内膜基质细胞2d或3d后,结果均显示,随着米非司酮浓度增加,基质细胞的细胞抑制率(IR)逐渐升高。米非司酮作用基质细胞2d后,50和100μmol/L组的IR值均高于对照组,差异均具有统计学意义(P<0.05);米非司酮作用3d后,25、50和100μmol/L组的IR值均高于对照组,差异均有统计学意义(P<0.05)。通过对IR值进行直线回归分析,结果表明:米非司酮处理2d和3d对原代人子宫内膜基质细胞的IC50分别为52.85μmol/L和86.46μmol/L。结论改良后的"二次消化法"可获得高纯度人子宫内膜基质细胞和上皮细胞。米非司酮对原代人子宫内膜基质细胞有抑制作用,并呈时间和剂量依赖效应。Objective：To explore the inhibitiory effect of mifepristone on the proliferation of human endometrial cells in primary culttures.Methods：Primary human endometrial cells were isolated by an improved ＂twice-enzyme digestion＂method.The purity of primary human endometrial stomal and epithelial cells was examined by flow cytometer（FCM）based on vimentin and cytokeration expression respectively.Human endometrial stomal cells treated with different concentrations（0,25,50 and 100μmol/L）of mifepristone were examined for the growth inhibition with MTT assay on day 2and day 3of mifepristone administration.The median effective inhibitory concentration 50%（IC50）was obtained.Results：Compared to primary human endometrial epithelial cells,primary human endometrial stormal cells grew more rapidly.FCM results demonstrated that a total purity of 97% and 90% was obtained in primary human endometrial stomal and epithelial cells,respectively.The inhibition rating of mifepristone on the cultured human endometrium stomal cells at day 2and day 3 were both increased in a dosedependent manner.Compared to that of control group,IR values were significantly increased in 50μmol/L and 100μmol/L groups on day 2（P〈0.05）,and significantly increased in 25μmol/L,50μmol/L and 100μmol/L groups on day 3（P〈0.05）.The IC50 of primary human endometrial stormal cells,by a linear regression analysis of IR values,were 52.85μmol/L on day 2and 86.46μmol/L on day 3respectively.Conclusions：An improved method ＂twice enzyme digestion＂ was established to obtained a high purity of human endometrial cells.Mifepristone inhibits the proliferation of cultrued human endometrial stormal cells in a dose-dependent manner.

关 键 词：米非司酮 人子宫内膜细胞 细胞增殖 半数抑制浓度 

分 类 号：R[医药卫生]