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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 delayed implantation mouse uterus and the regulation of steroid hormone on its expression. Methods: Kunming mice were used as experimental animals. Delayed implantation and activation models were established. Mouse ovariectomized steroid treatment model was established. Ezrin mRNA expression in mouse endometrium was detected by reverse transcription-polymerase chain reaction (RT-PCR) The immunohistochemical (IHC) ABC method was used to detect the localization of Ezrin in mouse uterus. Results: 1 Delayed implantation and activation of mouse uterus Ezrin expression: RT-PCR results show that delayed activation group mice uterine implantation Ezrin mRNA expression was significantly higher than delayed implantation (P <0.05). IHC results showed that weak Ezrin immunostaining was detectable in delayed implantation mouse uterine epithelium and stroma, and in delayed implantation activation mouse uterus implantation site, Ezrin immunostaining was mainly located below the implantation site In the matrix. Semi-quantitative immunohistochemical staining showed that the protein level of Ezrin in implantation-delayed mice was higher than that in delayed implantation group (P <0.05). 2 estrogen and progesterone on mouse uterus Ezrin expression regulation: ovariectomized mice estrogen and progesterone treatment, sesame oil group did not detect Ezrin mRNA or protein expression, and estrogen and progesterone can induce Ezrin expression. The mRNA expression of Ezrin in estrogen (E) group, progesterone (P) group and E + P group was significantly higher than that in E group (P <0.05) + P group, P group and E group Ezrin mRNA expression was significantly different (P <0.05). The result of IHC showed that the immunostaining of Ezrin in sesame oil group was negative. The immunostaining of Ezrin in E group was mainly located in endometrial luminal epithelium and glandular epithelium. E + P group and P group were detectable in luminal epithelium, glandular epithelium and stroma The results of semi-quantitative analysis showed that compared with sesame oil group, the protein levels of Ezrin in E group, E + P group and P group were significantly increased (P <0.05), E group was more significant (P < 0.01). CONCLUSION: It is speculated that the expression of Ezrin in mouse uterus may be regulated by estrogen and progesterone. Estrogen may induce the expression of Ezrin in mouse endometrial epithelium. Progesterone mainly promotes the expression of Ezrin in stromal cells.