埃兹蛋白在延迟着床小鼠子宫的表达及雌孕激素对其的调节作用

Expression of Ezrin in Mouse Endometrium Under Delayed Implantation and the Regulation of Ovarian Steroid Hormones on Its Expression

目的:探讨埃兹蛋白(Ezrin)在延迟着床小鼠子宫的表达及甾体激素对其表达的调节。方法:以昆明小鼠为实验对象,分别建立延迟着床与激活模型、小鼠卵巢切除类固醇激素处理模型,采用逆转录聚合酶链式反应(RT-PCR)法检测小鼠子宫内膜Ezrin mRNA的水平,免疫组织化学(IHC)ABC法检测小鼠子宫Ezrin的定位。结果:1延迟着床与激活小鼠子宫Ezrin的表达:RT-PCR结果显示,延迟着床激活组小鼠子宫着床点Ezrin的mRNA表达显著高于延迟着床组(P<0.05)。IHC结果显示,在延迟着床小鼠子宫腔上皮及基质中可检测到弱的Ezrin的免疫染色,在延迟着床激活组的小鼠子宫着床点,Ezrin的免疫染色主要位于着床位点下方的基质中。免疫组化染色半定量结果显示,延迟着床激活组小鼠子宫着床点Ezrin的蛋白水平较延迟着床组高(P<0.05)。2雌、孕激素对小鼠子宫Ezrin的表达调节:卵巢切除小鼠雌、孕激素处理后,芝麻油组未检测到Ezrin的mRNA或蛋白表达,而雌、孕激素均可诱导Ezrin的表达。RT-PCR结果显示,雌激素(E)组、孕激素(P)组和E+P组Ezrin的mRNA表达均升高(P<0.05),以E组更为显著(P<0.01),E+P组、P组和E组各组间Ezrin的mRNA表达有显著性差异(P<0.05)。IHC结果显示,芝麻油组Ezrin的免疫染色为阴性;E组Ezrin的免疫染色主要位于子宫内膜腔上皮和腺上皮;E+P组及P组在腔上皮、腺上皮和基质中均可检测到Ezrin蛋白的表达;半定量分析结果显示,与芝麻油组比较,E组、E+P组和P组小鼠子宫Ezrin的蛋白水平均升高(P<0.05),E组更为显著(P<0.01)。结论:推测小鼠子宫Ezrin的表达可能受雌孕激素的双重调节,雌激素可诱导小鼠子宫内膜上皮的Ezrin的表达,孕激素主要促进基质细胞Ezrin的表达。

Objective. To investigate the expression of ezrin in mouse endometrium under delayed im- plantation and steroid regulation on ezrin in mouse uterus. Methods. Immunohistochemistry and reverse transcription polymerase chain reaction （RT-PCR） were adopted to detect the localization and expression of ezrin in mouse uterus by using of delayed and activated implantation and ster- oids treatment models. Results： ①Expression of ezrin in delayed and activated implantation mod-el. RT-PCR result showed that the mRNA level of ezrin was significantly higher in activated mouse uterus than that of in delayed implantation group （P〈0.05）. Immunohistochemistry assay showed that ezrin protein was weakly detected in the luminal epithelium and stroma in delayed uterus, while strongly detected at the subluminal stroma surrounding implanted embryo, which was significantly higher than that in delayed implantation group by semi-quantitative analysis （P 〈0.05）. ②In ovariectomized mice, either mRNA or protein of ezrin was nearly undetectable in oil treated group （control）, whereas estrogen （E） and progesterone （P） could induce ezrin expression. RT-PCR results showed that ezrin mRNA level was higher in E, E＋P and P groups （P 〈0.05） when compared with control group （P〈0.01）, which was prominent in E group. Immunostaining found that ezrin was negative in control group, while was strong positve to luminal and glandular epithelium in endometrium in E group, and could be detected both in epithelium and stroma when treated with P or E＋P. Semi-quantitative analysis showed that the mean optical density value of immunostaining of ezrin was higher in E, E＋P and P group compared with control group （P〈0.05, G0. 05 and G0. 01, respectively）, which was prominent in E group. Conclusion： Estrogen and progesterone synergistically regulate ezrin expression. Estradiol can induce the expression of ezrin in endometrial epithelium, while progesterone mainly promotes the expression of ezrin in stroma.