摘要
目的探讨不同浓度细胞外信号调节激酶(ERK)抑制剂(U0126)对子宫内膜异位症(EMs)小鼠的治疗作用,并分析U0126选择性抑制雌二醇(E_(2))-雌激素受体/G蛋白耦联雌激素受体(ER/GPER)-ERK1/2通路对小鼠异位病灶增殖及血管新生的影响。方法建立EMs小鼠模型,将造模成功的小鼠随机分为模型对照组(注射生理盐水)、孕三烯酮组(给予孕三烯酮灌胃,0.325 mg/kg)、U0126低剂量组(腹腔注射U01260.5 mg/kg)、U0126中剂量组(腹腔注射U01261.0 mg/kg)和U0126高剂量组(腹腔注射U01262.0 mg/kg),并以正常小鼠设为空白对照组,每组5只。给药3周后统一处死小鼠,用酶联免疫吸附试验检测小鼠血清E_(2)的含量,测量异位病灶的大小并进行苏木素-伊红(HE)染色观察内膜结构,应用免疫组织化学法检测各组小鼠在位内膜和异位病灶组织中ER、GPER、β-ERK1/2、CD31和增殖细胞核抗原(PCNA)的定位和表达情况。结果模型对照组、不同U0126剂量组和孕三烯酮组小鼠血清E 2水平显著高于空白对照组(P均<0.05);模型对照组、不同U0126剂量组和孕三烯酮组间小鼠血清E_(2)水平比较差异无统计学意义(P>0.05)。各组小鼠药物治疗3周后,处死小鼠解剖后发现,不同U0126剂量组、孕三烯酮组的异位病灶体积显著小于模型对照组(P<0.05),且随着U0126剂量增加,病灶体积有逐渐减小的趋势。免疫组织化学结果显示,ER主要定位于上皮细胞和腺上皮细胞核,GPER主要定位于上皮细胞和腺上皮细胞膜,β-ERK1/2主要定位于上皮细胞和腺上皮细胞质,CD31主要定位于血管,PCNA主要定位于上皮细胞和腺上皮细胞核。结论EMs小鼠体内血清雌激素含量升高且ERs表达受雌激素调控;U0126能够抑制EMs小鼠异位病灶生长和腺体增殖,其机制可能与选择性抑制E_(2)-ER/GPER-ERK1/2通路活性、降低异位病灶生长增殖、抑制血管新生等有关。
Objective:To investigate the therapeutic effects of different concentrations of extracellular regulated protein kinase(ERK)inhibitor(U0126)in mice with endometriosis(EMs)and to analyze the effects of selective inhibition of the estradiol-estrogen receptor/G-protein-coupled estrogen receptor-extracellular regulated protein kinase 1/2(E_(2)-ER/GPER-ERK1/2)pathway by U0126 on the proliferation of ectopic foci and on the neovascularization of mice.Methods:The EMs mouse model was established,and the mice that successfully completed the procedure were randomly allocated to the model control group(injected with normal saline),gestrinone group(intragestric administration of 0.325 mg/kg of gestrinone),the U0126 low-dose group(intraperitoneal injection administration of 0.5 mg/kg of U0126),the U0126 medium-dose group(intraperitoneal injection administration of 1.0 mg/kg of U0126)and the U0126 high-dose group(intraperitoneal injection administration of 2.0 mg/kg of U0126),with a normal blank control group of 5 mice in each group.After 3 weeks of drug administration,the mice were uniformly executed,and the serum E_(2) level was detected by enzyme-linked immunosorbent assay,the size of ectopic lesions was measured and HE staining was carried out to observe the structure of the endometrium,and immunohistochemistry was applied to detect the localization and the expression of ER,GPER,β-ERK1/2,CD31,and proliferating nuclear antigen(PCNA)in the endometrium and ectopic lesions of the mice in each group.Results:The serum E_(2) levels in the model control group,different doses of U0126 groups and the gestrinone group were significantly higher than that of the blank control group(all P<0.05).There was no statistically significant difference in serum E_(2) level between the model control group,different doses of U0126 groups,and progesterone group(P>0.05).After 3 weeks of drug treatment in each group,after the mice were sacrificed and dissected,it was found that the ectopic lesion volumes in different dose of U0126 groups and the gestrinone group were significantly smaller than those in the model control group(P<0.05),and the lesion volumes gradually decreased with the increase of U0126 dose.Immunohistochemical results showed that ER was mainly localized in the nuclei of epithelial cells and glandular epithelial cells,GPER was mainly localized in the membranes of epithelial cells and glandular epithelial cells,β-ERK1/2 was mainly localized in the cytoplasm of epithelial cells and glandular epithelial cells,CD31 was mainly localized in blood vessels,and PCNA was mainly localized in the nuclei of epithelial cells and glandular epithelial cells.Conclusions:In EMs mice,serum estrogen level was elevated and the expression of ERs was regulated by estrogen.U0126 was able to inhibit the growth of ectopic foci and glandular proliferation in mice with endometriosis,and the mechanism may involve the selective inhibition of E_(2)-ER/GPER-ERK1/2 pathway activity,the reduction of ectopic foci growth and proliferation,and the inhibition of the development of neovascularization.
作者
肖文婷
韩明
程鑫
王桂丽
XIAO Wen-ting;HAN Ming;CHENG Xin;WANG Gui-li(Affiliated Hospital of Shandong Second Medical University,Weifang 261000;School of Basic Medical Sciences,Shandong Second Medical University,Weifang 261000;School of Medical Imaging,Shandong Second Medical University,Weifang 261000)
出处
《生殖医学杂志》
2025年第8期1087-1094,共8页
Journal of Reproductive Medicine
基金
山东省医药卫生科技发展计划项目(202005010475)。
关键词
子宫内膜异位症
细胞增殖
雌二醇
新生血管化
动物实验
Endometriosis
Cell proliferation
Estradiol
Neovascularization
Animal experimentation
