Progesterone regulating glutathione S-transferase Omega-1 expression in the mouse uterine luminal and glandular epithelium during preimplantation period

目的研究谷胱甘肽S-转移酶Omega-1(Gsto 1)在小鼠胚胎着床过程中的表达和孕酮的调节。方法105只CD1小鼠,分为正常妊娠模型和类固醇激素处理模型。正常妊娠模型中,收集妊娠第1~第5天子宫,采用Real-time PCR、原位杂交和Western blotting 3种方法检测Gsto1的表达变化;类固醇激素处理模型均采用卵巢切除2周后的小鼠,又分为雌孕激素处理组、孕酮处理不同时间组和孕酮受体拮抗剂Ru486处理组,所有组中的对照均用芝麻油处理。雌孕激素处理组中,收集芝麻油、雌激素、孕酮、雌激素加孕酮分别处理12h后的子宫;孕酮处理不同时间组中,收集芝麻油和孕酮分别处理1、3、12、24 h后的子宫;Ru486处理组中,收集芝麻油、Ru486、孕酮、Ru486加孕酮分别处理12 h后的子宫。类固醇激素处理模型使用Real-time PCR和Western blotting两种方法检测Gsto1的表达变化。结果 Gsto1主要在妊娠第1~4天的子宫腔上皮及腺上皮中表达,其中,妊娠第1~3天表达量较高,第4天表达量较低,第5天着床点和非着床点均不表达。孕酮诱导Gsto1的表达,雌激素不能诱导Gsto1的表达,并能抑制孕酮对Gsto1的诱导。Ru486降低孕酮对Gsto1的诱导,孕酮处理1、3、12 h均促进Gsto1的表达,但作用24 h后,抑制Gsto1的表达。结论 Gsto1在小鼠妊娠早期子宫腔上皮及腺上皮中表达,雌激素能够拮抗孕酮对Gsto1的诱导,孕酮可以通过孕酮受体调节Gsto1的表达,并且具有短时调节作用。Objective To investigate the expression and regulation of glutathione S-transferase Omega-1( Gsto1) in the mouse uterus during embryo implantation. Methods A total of 105 CD1 mice were divided into the normal pregnancy model and steriod hormone treatment model. Uterus were collected from days 1 to 5 of pregnancy in normal pregnancy model,and Gsto1 expression was detected by Real-time PCR,in situ hybridization and Western blotting. Ovariectomized mice were used in the steriod hormone model after 2 weeks,and divided into estrogen and progesterone treatment group,progesterone treatment course group,progesterone receptor antagonist Ru486 treatment group. Sesame oil was used for the control of all groups. In the estrogen and progesterone treatment group,uterus was collected after sesame oil,estrogen,progesterone,estrogen plus progesterone treatment 12 hours,respectively. In the progesterone treatment course group,uterus was collected after progesterone treatment 1 hours,3 hours,12 hours and 24 hours,respectively. In Ru486 treatment group,uterus was collected after sesame oil,Ru486,progesterone,Ru486 plus progesterone treatment 12 hours. Gsto1 expression was detected by Real-time PCR,and Western blotting in the steriod hormone model. Results Gsto1 was mainly expressed in the uterine luminal and glandular epithelium on days 1 to 4 of pregnancy. Gsto1 expression was high on day 1 to 3,but became lower onday 4. On day 5,Gsto1 expression was not detected at implantation sites and non-implantation sites. Progesterone induced Gsto1 expression. Estrogen did not induce Gsto1 expression,but inhibited the induction of progesterone on Gsto1. Ru486 reduced the induction of progesterone on Gsto1. Progesterone treatment for 1 hour,3 hours,12 hours promoted Gsto1 expression,but after 24 hours,inhibited Gsto1 expression. Conclusion This study suggests that Gsto1 is mainly expressed in the uterine luminal and glandular epithelium during preimplantation. Estrogen inhibits the induction of progesterone on Gsto1.Progesterone enhances Gsto1 expression by progesterone receptor in short time.国家自然科学基金(31401953);; 安徽省教育厅自然科学重点项目(2014A143