子宫内膜异位症大鼠在位及异位内膜组织中RAS与MEK的表达

Expression of RAS and MEK in the eutopic and ectopic endometrial tissues of rats with endometriosis

目的探讨子宫内膜异位症大鼠在位及异位内膜组织中原癌基因蛋白(RAS)与丝裂原活化蛋白激酶(MEK)的表达水平。方法选取SPF级健康雌性未孕SD大鼠20只,自体移植法建立EMs大鼠模型,其中假手术组作为对照组(每组10只)。应用免疫组织化学法、Western-blot及qRT-PCR检测法检测EMs大鼠在位及异位内膜组织中RAS与MEK蛋白及mRNA表达情况,Pearson相关分析分析两种蛋白表达的相关性。结果苏木素伊红染色显示异位病灶组织见腺体及间质即为模型成功;免疫组化结果显示:RAS蛋白主要定位于子宫内膜腺上皮及腺体的胞浆中,间质内少量表达;MEK主要定位于子宫内膜腺体的胞浆中,胞核中少量表达。Western-blot及qRT-PCR结果显示:与对照组相比,EMs模型组在位及异位内膜组织中RAS、MEK蛋白及mRNA的表达均明显升高,异位内膜组中的表达高于在位内膜组,各组间差异具有统计学意义(P<0.05);Pearson相关分析结果显示,EMs大鼠在位及异位内膜组织中RAS与MEK蛋白表达呈正相关(r_(异位内膜组)=0.68,P<0.05;r_(在位内膜组)=0.79,P<0.05)。结论EMs大鼠在位及异位内膜组织中RAS与MEK蛋白表达升高且呈正相关,提示两者表达相辅相成,可能与子宫内膜异位症的发生密切相关。

Objective To investigate the expression of proto-oncogene protein(RAS)and mitogen-activated protein kinase(MEK)in the eutopic and ectopic endometrial tissues of rats with endometriosis.Methods Twenty SPF healthy female non-pregnant SD rats were selected,and EMs rat models were established with autologous transplantation.Among them,the rats received sham operation were used as the control group(ten rats).Immunohistochemistry,Western-blot,and q RT-PCR were used to detect the expression of RAS and MEK protein and m RNA in eutopic and ectopic endometrial tissues of EMs rats.Results HE staining showed glands and stroma in ectopic lesions,which indicated the models were successfully established.The results of immunohistochemistry showed that the RAS protein was mainly located in the cytoplasm of endometrial glandular epithelium and glands,with a small amount of expression in the interstitium;MEK was mainly located in the cytoplasm of endometrial glands,with a small amount of expression in the nucleus.The results of Western-blot and q RT-PCR showed that compared with the control group,the protein and m RNA expression of RAS and MEK was significantly increased in the EMs model group,and the expression in the ectopic endometrium group was higher than that in the eutopic endometrium group,with statistically significant differences(P<0.05).Pearson correlation analysis showed that RAS and MEK proteins were positively correlated in the EMs rats and in the ectopic endometrium(rectopic endometrium group=0.68,P<0.05;reutopic endometrium group=0.79,P<0.05).Conclusion The expressions of RAS and MEK protein in eutopic and ectopic endometrial tissues of EMs rats are elevated and positively correlated,suggesting that the expressions of the two complement each other and may be closely related to the occurrence of endometriosis.