促性腺激素释放激素拮抗剂上调小鼠种植窗期子宫自然杀伤细胞比例并增强其毒性

Gonadotropin-releasing hormone-antagonist up-regulate the proportion of uterine natural killer cells and enhance their toxicity during the implantation window period of mice

目的·探讨促性腺激素释放激素拮抗剂(gonadotropin-releasing hormone-antagonist,GnRH-ant)对小鼠种植窗期子宫自然杀伤细胞(uterine natural killer cell,uNK细胞)的比例及毒性的影响。方法·将16只C57BL/6小鼠随机分为GnRH-ant组和对照组,每组8只。自动情期第3日起,每日向GnRH-ant组腹腔注射GnRH-ant(1.5μg/100 g,连续7 d),对照组于相同时间点注射等体积生理盐水。第7日,2组小鼠均注射人绝经期尿促性腺激素(40 U/100 g),次日注射人绒毛膜促性腺激素(100 U/100 g)并于48 h后处死小鼠,取出子宫组织进行原代消化获取单细胞悬液。采用流式细胞术分析uNK细胞的比例,及毒性分子穿孔素(perforin,Pf)和颗粒酶B(granzyme B,Gz-B)的表达水平。结果·与对照组相比,GnRH-ant组小鼠种植窗期uNK细胞的比例增加(P=0.000)、增殖水平升高(P=0.000)、凋亡水平降低(P=0.004),且该组小鼠的uNK细胞毒性分子Pf(P=0.000)和Gz-B(P=0.034)的表达均上调。结论·GnRH-ant可上调小鼠种植窗期uNK细胞的比例并增强其毒性。

Objective·To investigate the effects of gonadotropin-releasing hormone-antagonist(GnRH-ant)on the proportion and toxicity of mice uterine nature killer(uNK)cells during implantation window.Methods·Sixteen C57BL/6 mice were randomly divided into GnRH-ant group and control group,with 8 mice in each group.From the 3rd day of the estrous cycle,GnRH-ant(1.5μg/100 g)was injected intraperitoneally into the mice of the GnRH-ant group for 7 days continuously,and the control group was injected with the same volume of normal saline at the same time point.On the 7th day,the mice of the two groups were injected with human menopausal gonadotropin(40 U/100 g).The next day,they were injected with human chorionic gonadotropin(100 U/100 g)and sacrificed after 48 h.The uterus tissues were taken out for primary digestion to obtain single-cell suspension.Flow cytometry was used to analyze the proportion of uNK cells and the expression levels of toxicity molecules perforin(Pf)and granzyme B(Gz-B).Results·Compared with the control group,the proportion of uNK cells in GnRH-ant group increased(P=0.000),the proliferation level increased(P=0.000),the apoptosis level decreased(P=0.004),and the expression of toxicity molecules Pf(P=0.000)and Gz-B(P=0.034)were up-regulated.Conclusion·GnRH-ant may upregulate the proportion of uNK cells and enhance their toxicity in the implantation window period of mice.