EMs异位及在位内膜间质细胞Cx43的表达及GJIC功能的体外研究

Study on Cx43 expression and function of GJIC in ectopic and eutopic endometrial stromal cells in vitro

目的:研究子宫内膜异位症(EMs)的异位与在位内膜间质细胞中Cx43的表达及间隙连接细胞间通讯(GJIC)功能,探讨间质细胞GJIC功能异常对EMs发病的影响。方法:取24例卵巢处子宫内膜异位灶、41例EMs在位内膜以及30例正常子宫内膜,分离出间质细胞,加入雌、孕激素培养,建立体外间质细胞培养模型。利用激光共聚焦显微镜(LSCM)分别检测三组间质细胞的Cx43表达及GJIC功能。结果:间质细胞成功培养率分别为:异位内膜组45.8%(11/24)、EMs在位内膜组92.7%(38/41)、对照组93.3%(28/30),异位内膜间质细胞纯度为95%,在位内膜间质细胞纯度达98%。EMs异位内膜间质细胞中Cx43表达水平及GJIC功能明显比其他两组低,正常对照组的Cx43表达水平及GJIC功能最高,且差异均有显著性(P﹤0.01)。结论:(1)同时建立EMs异位内膜、在位内膜及正常内膜间质细胞的体外模型,有助于研究EMs的发病机制;(2)间质细胞中Cx43蛋白表达下调及其GJIC功能异常与EMs发生有关,调节Cx43表达或GJIC功能可能是潜在的治疗策略。

Objective：To investigate Cx43 expression and gap junctional intercellular communication （GJIC） in ectopic and eutopic endometrial stromal cells in endometriosis （EMs）,and to explore the influence of aberrant GJIC in stromal cells on pathogenesis of EMs. Methods：The stromal cells were isolated from samples including 24 ectopic endometriotic tissues located in ovaries,41 eutopic endometria with endometriosis and 30 normal endometria. The endometrial stromal cells models were established in vitro by being cultured in manual conditions mimicked with estrogen and progesterone. Laser scanning confocal microscopy （LSCM） was used to determine the expression of Cx43 protein and the function of GJIC in three groups stromal cells. Results：The success rate of isolation and culture of endometriotic stromal cells was 45.8%（11/24） ;of eutopic endometrial stromal cells with EMs was 92.7 （38/41）;of normal endometrial stromal cells was 93.3%（28/30）. The purities of ectopic and eutopic endometrial stromal cells were 95% and 98% respectively. The level of Cx43 protein and the function of GJIC in stromal cells from ectopic endometrial tissues were much lower than those from the other two groups, the highest level of Cx43 protein and the function of GJIC were observed in normal endometrial stromal cells group,and the differences among these groups were significant （P 〈 0.01 ）. Conclusions： （1） It will be helpful to establish models of normal, ectopic and eutopic endometrial stromal cells in vitro simultaneously when investigating the pathgenesis of EMs. （2）Downregulation of Cx43 expression and aberrant function of GJIC are related to pathogenesis of EMs. Regulation of Cx43 or GJIC in endometrial stromal cells is implied to be a potential strategy to treat EMs.