The Influence of <italic style="font-style: italic">Shoutai Wan </italic>（寿胎丸） on Lactic Acid Content at the Maternal-Fetal Interface and Expression of Immune-Related Factors in Recurrent Miscarriage Model Mice

LYU Jingfang; JIANG Min; ZHOU Zhenzhen; ZHANG Li; WANG Shuhui; LI Dandan; SONG Yajing; DU Huilan

doi:10.13288/j.11-2166/r.2024.09.011

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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

更新时间：2024-04-25

- 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
- Journal of Traditional Chinese Medicine Vol. 65, Issue 9, Pages: 934-942(2024)
- 作者机构：
  
  1.河北中医药大学，河北省石家庄市桥西区新石南路326号，050091
  2.河北省中西医结合生殖疾病协同创新中心
  3.河北省中西医结合肝肾病证研究重点实验室
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2024.09.011
  CLC：
- Received：16 December 2023，
  
  Revised：2024-01-20，
  
  Published：02 May 2024
- 稿件说明：
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吕竞芳,蒋敏,周真真等.寿胎丸对复发性流产模型小鼠母胎界面乳酸含量及免疫相关因子表达的影响[J].中医杂志,2024,65(09):934-942.

LYU Jingfang,JIANG Min,ZHOU Zhenzhen,et al.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[J].Journal of Traditional Chinese Medicine,2024,65(09):934-942.
吕竞芳,蒋敏,周真真等.寿胎丸对复发性流产模型小鼠母胎界面乳酸含量及免疫相关因子表达的影响[J].中医杂志,2024,65(09):934-942. DOI： 10.13288/j.11-2166/r.2024.09.011.

LYU Jingfang,JIANG Min,ZHOU Zhenzhen,et al.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[J].Journal of Traditional Chinese Medicine,2024,65(09):934-942. DOI： 10.13288/j.11-2166/r.2024.09.011.

摘要

目的

从母胎界面酸性微环境下免疫耐受角度探讨寿胎丸治疗复发性流产（RSA）的可能机制。

方法

雌性CBA/J小鼠随机分为正常组、模型组、地屈孕酮组、寿胎丸组，每组15只。正常组和模型组小鼠给予0.2 ml/d蒸馏水灌胃，寿胎丸组给予寿胎丸水煎液0.15 g/（10 g·d）灌胃，地屈孕酮组给予地屈孕酮片0.44 mg/（10 g·d）灌胃。灌胃14天后，正常组CBA/J雌性小鼠与BALB/c雄性小鼠2∶1合笼，其余各组CBA/J雌性小鼠与DBA/2雄性小鼠2∶1合笼建立RSA小鼠模型。次日雌性小鼠清晨做阴道涂片，以阴道涂片出现大量精子并见到阴道栓作为妊娠第1天。见栓后按照之前的给药方式继续给药，直至妊娠第10天。各组雌鼠在妊娠第10天取母胎界面组织，乳酸脱氢酶比色法检测乳酸（LA）含量；qPCR法、Western blot法分别检测免疫相关因子白细胞介素4（IL-4）、γ干扰素（IFN-γ）、转化生长因子β1（TGF-β1）、叉头转录因子（Foxp3）mRNA和蛋白表达；流式细胞术检测辅助性T淋巴细胞1（Th1）、辅助性T淋巴细胞2（Th2）、调节性T细胞（Treg）、经典巨噬细胞（M1）、替代巨噬细胞（M2）的数量。采用双变量Pearson检验分析LA含量与Th1、Th2、Treg、M1、M2数量相关性，以及LA含量与IL-4、IFN-γ、TGF-β1、Foxp3蛋白和mRNA的相关性。

结果

妊娠第10天，与正常组比较，模型组LA含量降低，母胎界面组织中IL-4、TGF-β1、Foxp3蛋白及mRNA表达降低，IFN-γ蛋白及mRNA表达升高，Th1和M1细胞数量增多，Th2、Treg、M2细胞数量减少（

＜0.05或

＜0.01）。与模型组比较，寿胎丸组及地屈孕酮组LA含量升高，母胎界面组织中IL-4、TGF-β1、Foxp3蛋白及mRNA表达升高，IFN-γ蛋白及mRNA表达降低，Th1和M1细胞数量减少，Th2、Treg和M2细胞数量增多（

＜0.05或

＜0.01）。LA含量与Th2、Treg、M2细胞数量，IL-4、TGF-β1、Foxp3蛋白及mRNA表达呈正相关（

＜0.05或

＜0.01）；LA含量与Th1、M1细胞数量，IFN-γ蛋白及mRNA表达呈负相关（

＜0.05或

＜0.01）。

结论

寿胎丸可能通过调控RSA模型小鼠母胎界面酸性微环境中免疫相关因子的表达，从而提高免疫耐受，发挥其保胎作用。

Abstract

Objective

To explore the possible mechanisms of

Shoutai Wan

（寿胎丸） in treating recurrent miscarriage （RSA） from the perspective of immune tolerance under the acidic microenvironment at the maternal-fetal interface.

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 spermatozoa 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 dehydrogenase 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-γ）， transforming 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）， classical macrophage （M1）， and alternative macrophage （M2）. The bivariate Pearson test was used to analyze the correlation 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-γ， 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-γ protein and mRNA increased. The numbers of Th1 and M1 cells increased， while the numbers of Th2， Treg， and M2 cells decreased （

＜0.05 or

＜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-γ protein and mRNA decreased. The numbers of Th1 and M1 cells decreased， while the numbers of Th2， Treg， and M2 cells increased （

＜0.05 or

＜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 protein， and mRNA （

＜0.05 or

＜0.01）； the LA content was negatively correlated with the numbers of Th1， M1 cells， and the expression of IFN-γ protein and mRNA （

＜0.05 or

＜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.

关键词

Keywords

references

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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

DOI：10.13288/j.11-2166/r.2024.09.011

摘要

Abstract

关键词

Keywords

references