基于Nrf2/HO-1信号通路的寿胎丸对复发性流产大鼠的影响

Effects of Shoutai Pills on Recurrent Spontaneous Abortion Mice Based on Nrf2/HO-1 Signaling Pathway

目的观察寿胎丸对复发性流产(RSA)大鼠的影响,从Nrf2/HO-1信号通路探讨其作用机制。方法32只SD雌鼠与16只SD雄鼠合笼饲养过夜,将32只孕鼠随机分为正常妊娠组、模型组、地屈孕酮组及寿胎丸组。除正常妊娠组外,其余各组在妊娠第1~9日下午予羟基脲溶液灌胃,妊娠第10日上午予米非司酮溶液灌胃建立RSA大鼠模型。寿胎丸组第1~9日上午予寿胎丸药液灌胃,地屈孕酮组予地屈孕酮溶液灌胃,模型组和正常妊娠组予等量生理盐水灌胃。米非司酮灌胃2 h后分离子宫,计算子宫脏器指数和胚胎丢失率,HE染色观察蜕膜组织病理变化,ELISA检测血清活性氧(ROS)含量,Western blot检测蜕膜组织核转录因子E2相关因子2(Nrf2)、血红素氧合酶1(HO-1)、醌氧化还原酶1(NQO1)蛋白表达,RT-qPCR检测蜕膜组织Nrf2、HO-1、NQO1 mRNA表达。结果与正常妊娠组比较,模型组大鼠子宫脏器指数显著降低,胚胎丢失率显著升高(P<0.01);蜕膜细胞胞浆水肿,细胞核萎缩或消失;血清ROS含量显著增加(P<0.01),蜕膜组织Nrf2、HO-1、NQO1蛋白和mRNA表达显著降低(P<0.01,P<0.05)。与模型组比较,寿胎丸组大鼠子宫脏器指数显著升高,胚胎丢失率显著降低(P<0.05);蜕膜细胞胞浆水肿、核仁固缩明显改善;血清ROS含量显著减少(P<0.01),蜕膜组织Nrf2、HO-1、NQO1蛋白和mRNA表达显著升高(P<0.05,P<0.01)。结论寿胎丸可能通过激活Nrf2/HO-1通路提高RSA大鼠抗氧化应激能力,从而治疗RSA。

Objective To observe the effects of Shoutai Pills on rats with recurrent spontaneous abortion(RSA);To explore its mechanism from the Nrf2/HO-1 signaling pathway.Methods Totally 32 SD female rats were co-caged with 16 SD male rats overnight.32 pregnant rats were randomly divided into normal pregnancy group,model group,dydrogesterone group and Shoutai Pills group.Except for the normal pregnancy group,the other groups were given hydroxyurea solution by gavage on the afternoon of the 1st to 9th day of pregnancy,and mifepristone solution was given by gavage on the morning of the 10th day of pregnancy to establish the RSA rat model.In addition,on the morning of the 1st to 9th day of pregnancy,the Shoutai Pills group was given Shoutai Pills liquid by gavage,the dydrogesterone group was given dydrogesterone aqueous solution by gavage,the model group and the normal pregnancy group were given the same amount of saline by gavage.The uterus was isolated 2 h after mifepristone gavage,the uterine organ index and embryo loss rate were calculated,HE staining was used to observe the pathological changes in decidual tissue,ROS content in serum were detected by ELISA,the proteins expression of Nrf2,HO-1,and NQO1 in decidual tissue were detected by Western blot,and the mRNA expression of Nrf2,HO-1and NQO1 in decidual tissue were detected by RT-qPCR.Results Compared with the normal pregnancy group,the uterine organ index of rats in the model group decreased,and the embryo loss rate increased(P<0.01),the decidual cell cytoplasmic edema,and the nucleus atrophied or disappeared;the content of serum ROS significantly increased(P<0.01),the expression of Nrf2,HO-1,NQO1 protein and mRNA expression in decidual tissue significantly decreased(P<0.01,P<0.05).Compared with the model group,the uterine organ index of rats in the Shoutai Pills group increased,and the embryo loss rate decreased(P<0.05),decidual cell cytoplasmic edema and nucleolar pyknosis were significantly improved;the content of serum ROS significantly reduced(P<0.01),the expressions of Nrf2,HO-1,and NQO1 protein and mRNA in decidual tissue significantly increased(P<0.05,P<0.01).Conclusion Shoutai Pills may improve the anti-oxidative stress ability of RSA rats by activating the Nrf2/HO-1 pathway,thereby treating RSA.