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目的探讨干扰素/维甲酸联合应用诱导细胞凋亡相关基因-19(GRIM-19)在小鼠围着床期子宫内膜中的表达及其与胚胎着床的关系。方法收集早期妊娠第1~6天小鼠子宫组织,采用免疫组织化学法检测GRIM-19蛋白在子宫内膜上皮细胞中的表达及定位情况;收集早期妊娠、假孕和雌孕激素处理小鼠子宫组织,采用Western blotting和Real-time PCR检测GRIM-19蛋白和mRNA水平;TUNEL方法检测早期妊娠第1~6天小鼠子宫组织细胞凋亡情况;采用p EGFP GRIM-19质粒GRIM-19-siRNA转染技术使RL95-2细胞系过表达或低表达GRIM-19,检测其对RL95-2-Be Wo共培养体外着床模型球状体黏附率的影响,Annexin V/PI双染色法检测细胞凋亡,线粒膜电位检测试剂盒(JC-1)检测线粒体跨膜电位;采用Western blotting和Real-time PCR检测转染后信号转导子和转录激活因子3(STAT3)、肿瘤坏死因子(TNF)-α及白细胞介素(IL)-11蛋白和mRNA水平。结果 GRIM-19在早期妊娠第1~6天小鼠子宫腔上皮和腺上皮均有表达,妊娠第4天GRIM-19蛋白和mRNA水平减低,细胞凋亡减少;假孕第1~6天小鼠子宫内膜GRIM-19表达无明显差异;雌激素处理组小鼠子宫内膜GRIM-19表达减弱;过表达GRIM-19后,RL95-2-Be Wo共培养球状体黏附率降低,细胞凋亡增加,线粒体膜电位减低,RL95-2细胞系中STAT3及IL-11蛋白和mRNA水平减低,TNF-α蛋白和mRNA水平升高。结论 GRIM-19在胚胎着床中发挥重要作用,可能是通过调节细胞凋亡和免疫耐受为胚胎着床提供保障。
Objective To investigate the expression of GRIM-19 induced by interferon / retinoic acid in peri-implantation endometrium and its relationship with embryo implantation. Methods The uterine tissues of mice were collected from the first to the sixth day of pregnancy. The expression and localization of GRIM-19 protein in the endometrial epithelial cells were detected by immunohistochemical staining. The early pregnancy, pseudopregnancy and estrogen-progesterone-treated mice The expression of GRIM-19 protein and mRNA was detected by Western blotting and Real-time PCR. The apoptosis of mouse uterine tissue was detected by TUNEL method. The expression of GRIM- siRNA transfected RL95-2 cell line overexpression or low expression of GRIM-19, RL95-2-Be Wo to detect the impact of co-cultured in vitro implantation model of spherical adhesion rate, Annexin V / PI double staining of cells (JC-1) was used to detect mitochondrial transmembrane potential. Western blotting and Real-time PCR were used to detect the expression of signal transducer and activator of transcription 3 (STAT3), tumor necrosis factor TNF) -alpha and interleukin (IL) -11 protein and mRNA levels. Results GRIM-19 was expressed in the uterine epithelium and glandular epithelium of mice on the 1st to 6th day of gestation. The GRIM-19 protein and mRNA levels were decreased on the 4th day of gestation, and the apoptosis of GRIM-19 was decreased. On the 1st to 6th day of pregnancy, The expression of GRIM-19 in endometrium of mice was not significantly different. The expression of GRIM-19 in endometrium of estrogen-treated mice was weakened. The adhesion rate of RL95-2-Be Wo spheroid decreased after GRIM-19 was overexpressed, Increased death, decreased mitochondrial membrane potential, RL95-2 cell line STAT3 and IL-11 protein and mRNA levels decreased, TNF-α protein and mRNA levels. Conclusion GRIM-19 plays an important role in embryo implantation, which may provide a guarantee for embryo implantation by regulating apoptosis and immune tolerance.