孕激素对人子宫内膜间质细胞moesin表达的影响

Expression of moesin in Human Endometrial Stromal Cells Regulated by Progesterone

目的:探讨孕激素(P)对人子宫内膜间质细胞(hESC)中细胞骨架调节蛋白moesin的调节作用,为胚胎着床提供理论依据。方法:从因子宫肌瘤行全子宫切除术的妇女子宫标本中获取分泌期子宫内膜组织,分离、鉴定后建立人类子宫内膜间质细胞(hESC)体外模型。根据细胞培养液中添加激素不同分组为空白对照组(A组)、孕激素组(B组,P 10-5 mol/L)、雌、孕激素组(C组,E210-8 mol/L+P 10-5 mol/L)及米非司酮联合雌、孕激素组(D组,E210-8 mol/L+P 10-5mol/L+Mif 10-5 mol/L),培养3 d后应用细胞化学免疫方法和免疫印迹方法检测hESC moesin蛋白定位和定量的变化。结果:细胞化学荧光检测显示moesin主要分布于hESC的细胞胞质及胞膜突起、皱褶、细胞间接触等特殊细胞皮质膜部位,各实验组和对照组moesin定位无显著变化。免疫印迹方法显示B组、C组hESC中moesin表达显著均高于A组(P<0.05);B组和C组间表达无显著差异(P>0.05);D组hESC中moesin蛋白较C组表达显著降低(P<0.05)。结论:孕激素促进hESC的moesin表达上调,进一步通过调节细胞骨架重塑介导胚胎着床;米非司酮通过拮抗孕激素作用,抑制hESC moesin上调。

Objective： To investigate the expression of moesin in the human endometrial stromal cells（hESC） regulated by progesterone.Methods： Acquired the secretory endometrium from the patients undergoing hysterectomy for leiomyoma.Isolated and purified the hESC and cultured the hESC with the medium containing 0 mol/L progesterone（group A,the blank control）,10^-5 mol/L progesterone（group B）,10^-8 mol/L estradiol plus 10^-5 mol/L progesterone（group C）,10^-5 mol/L mifepristone combined with 10^-8 mol/L estradiol plus 10^-5 mol/L progesterone（group D）,respectively for 3 consecutive days.The expression of moesin was detected by cytochemistry and Western blotting.Results： Moesin wase mainly distributed in the cytoplastic and special cortical membranes such as membrane protuberances,ruffles and intercellular contact areas in hESC.Compared with the control,the distrubition of moesin in other 3 treatment groups didn＇t show differences using the fluorescence immunohistochemisty.Moesin expression was found significantly enhanced in group B than that in the control using Western blotting（P〈0.05）.Moesin expression was also significantly enhanced in group C（P〈0.05）.There was no difference between groups B and C（P〈0.05）.The expressin of moesin in group B was lower than that in group C（P〈0.05）.Conclusion： Moesin expression was increased by progesterone in hESC and mifepristone reversed the role of progesterone,which presumed moesin may be involved in implantation by regulating cytoskeleton reorganization.