The Influence of Shoutai Wan(寿胎丸)on Lactic Acid Content at the Maternal-Fetal Interface and Expression of Immune-Related Factors in Recurrent Miscarriage Model Mice
Objective To explore the possible mechanisms of Shoutai Wan(寿胎丸)in treating recurrent miscar-riage(RSA)from the perspective of immune tolerance under the acidic microenvironment at the maternal-fetal inter-face.Methods Female CBA/J mice were randomly divided into normal group,model group,progesterone group,and Shoutai Wan group,with 15 mice in each group.The mice in the normal group and model group were given 0.2 ml distilled water by gavage each day,the Shoutai Wan group given Shoutai Wan decoction 0.15 g/(10 g-d)by gavage,the progesterone group given progesterone tablets 0.44 mg/(10 g·d)by gavage.After gavage for 14 days,the mice were cohabited.Female CBA/J mice in the normal group were mated with male BALB/c mice at a ratio of 2∶1,and female CBA/J mice in the other groups were mated with male DBA/2 mice at a ratio of 2∶1 to establish the RSA mouse model.Vaginal smears were taken from the female mice the next morning,and the appearance of a large number of sper-matozoa and the presence of a vaginal plug were considered as the first day of pregnancy.After the appearance of the plug,the mice were continued to be administered according to the previous method until the 10th day of pregnancy.On the 10th day of pregnancy,maternal-fetal interface tissues were collected from each group of mice,and lactate de-hydrogenase colorimetric method was used to detect lactate(LA)content;qPCR method and Western blot method were used to detect the expression of immune-related factors interleukin-4(IL-4),interferon-gamma(IFN-y),transform-ing growth factor beta 1(TGF-β1),and forkhead box protein 3(Foxp3)mRNA and protein;flow cytometry was used to detect the numbers of helper T lymphocyte 1(Th1),helper T lymphocyte 2(Th2),regulatory T cell(Treg),clas-sical macrophage(M1),and alternative macrophage(M2).The bivariate Pearson test was used to analyze the corre-lation between LA content and the numbers of Th1,Th2,Treg,M1,and M2 cells,as well as the correlation between LA content and the expression of IL-4,IFN-y,TGF-β1,Foxp3 protein,and mRNA.Results On the 10th day of pregnancy,compared with the normal group,the LA content decreased in the model group,and the expression of IL-4,TGF-β1,Foxp3 protein and mRNA in the maternal-fetal interface tissues decreased,while the expression of IFN-γprotein and mRNA increased.The numbers of Th1 and M1 cells increased,while the numbers of Th2,Treg,and M2 cells decreased(P＜0.05 or P＜0.01).Compared with the model group,the LA content increased in the Shoutai Wan group and progesterone group.The expression of IL-4,TGF-β1,Foxp3 protein and mRNA in the maternal-fetal interface tissues increased,while the expression of IFN-y protein and mRNA decreased.The numbers of Th1 and M1 cells decreased,while the numbers of Th2,Treg,and M2 cells increased(P＜0.05 or P＜0.01).The LA content was positively correlated with the numbers of Th2,Treg,and M2 cells,and the expression of IL-4,TGF-β1,Foxp3 pro-tein,and mRNA(P＜0.05 or P＜0.01);the LA content was negatively correlated with the numbers of Th1,M1 cells,and the expression of IFN-γ protein and mRNA(P＜0.05 or P＜0.01).Conclusion Shoutai Wan may improve immune tolerance by regulating the expression of immune-related factors in the acidic microenvironment at the maternal-fetal interface of RSA model mice,thereby exerting its role in preventing miscarriage.
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