寿胎丸对复发性流产小鼠胚胎滋养细胞上皮间质转化的影响

唐思玲; 梁潇; 李荣霞; 宿滋; 刘雪平; 杜惠兰; 段彦苍

doi:10.3969/j.issn.1006-2157.2023.08.012

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寿胎丸对复发性流产小鼠胚胎滋养细胞上皮间质转化的影响

实验研究 | 更新时间：2023-09-08

- 寿胎丸对复发性流产小鼠胚胎滋养细胞上皮间质转化的影响
- Effects of Shoutai Pills on EMT of embryonic trophoblasts in recurrent spontaneous abortion mice
- 北京中医药大学学报 2023年46卷第8期页码：1139-1149
- 作者机构：
  
  1.河北中医药大学中西医结合学院　石家庄　050091
  2.河北中医药大学基础医学院
  3.河北省中西医结合生殖疾病协同创新中心
  4.河北省中西医结合肝肾病证研究重点实验室
- 作者简介：
  
  唐思玲，女，在读硕士生
  # 段彦苍，女，博士，教授，博士生导师，主要研究方向：中西医结合生殖内分泌，E-mail：duanyancang@163.com
- 基金信息：
  
  国家自然科学基金项目(U21A20403)
- DOI：10.3969/j.issn.1006-2157.2023.08.012
  中图分类号： R274.1
- 收稿日期：2022-09-15，
  
  网络出版日期：2023-05-12，
  
  纸质出版日期：2023-08-30
- 稿件说明：
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唐思玲, 梁潇, 李荣霞, 等. 寿胎丸对复发性流产小鼠胚胎滋养细胞上皮间质转化的影响[J]. 北京中医药大学学报, 2023,46(8):1139-1149.

TANG Siling, LIANG Xiao, LI Rongxia, et al. Effects of Shoutai Pills on EMT of embryonic trophoblasts in recurrent spontaneous abortion mice[J]. Journal of beijing university of traditional chinese medicine, 2023, 46(8): 1139-1149.
唐思玲, 梁潇, 李荣霞, 等. 寿胎丸对复发性流产小鼠胚胎滋养细胞上皮间质转化的影响[J]. 北京中医药大学学报, 2023,46(8):1139-1149. DOI： 10.3969/j.issn.1006-2157.2023.08.012.

TANG Siling, LIANG Xiao, LI Rongxia, et al. Effects of Shoutai Pills on EMT of embryonic trophoblasts in recurrent spontaneous abortion mice[J]. Journal of beijing university of traditional chinese medicine, 2023, 46(8): 1139-1149. DOI： 10.3969/j.issn.1006-2157.2023.08.012.

摘要

目的

探讨寿胎丸对复发性流产(RSA)小鼠胚胎滋养细胞上皮间质转化(EMT)及胚胎丢失率的影响。

方法

将90只6～8周龄CBA/J雌性小鼠按随机数字表法分为正常组、模型组、地屈孕酮组和寿胎丸低、中、高剂量组，每组15只；正常组与BALB/c雄鼠合笼建立正常妊娠小鼠，其他组雌鼠与DBA/2雄鼠合笼建立RSA小鼠模型。合笼前14 d开始每日给药，寿胎丸低、中、高剂量组分别以寿胎丸7.58、15.17、30.33 g/kg灌胃，地屈孕酮组以地屈孕酮0.003 g/kg灌胃，正常组和模型组以等体积蒸馏水灌胃。连续给药至妊娠第6天(PD 6)，各组随意选取9只孕鼠麻醉处死后剥离胚胎，HE染色观察PD 6小鼠母胎界面滋养细胞病理形态学改变；免疫荧光染色、实时荧光定量PCR检测各组胚胎滋养细胞EMT标志物上皮钙黏素(E-cadherin)、波形蛋白(Vimentin)及β－连环蛋白(β-catenin)的蛋白及mRNA表达。各组其余孕鼠继续给药至妊娠第14天(PD 14)，麻醉处死后观察胚胎丢失情况。

结果

与正常组比较，模型组孕鼠胚胎滋养细胞中上皮细胞标记物E-cadherin蛋白及mRNA表达升高(

0.05)，间质细胞标志物Vimentin蛋白及mRNA表达下降(

0.05)、β-catenin蛋白及mRNA表达下降(

0.05)，胚胎丢失率升高(

0.05)；与模型组比较，各剂量寿胎丸组及地屈孕酮组胚胎滋养细胞中E-cadherin蛋白及mRNA表达下调(

0.05)，Vimentin蛋白及mRNA表达升高、β-catenin蛋白及mRNA表达升高(

0.05)，其中寿胎丸中、高剂量组及地屈孕酮组更为明显，寿胎丸低剂量组胚胎丢失率有所降低(

0.05)，寿胎丸中、高剂量组及地屈孕酮组胚胎丢失率下降(

0.05)。

结论

寿胎丸能降低RSA小鼠胚胎丢失率，其机制可能是通过激活滋养细胞内β-catenin信号分子，促进EMT改善胚胎滋养细胞迁移侵袭能力，以利于胚胎在宫腔内着床而发挥保胎作用。

Abstract

Objective

We aimed to explore the effect of

Shoutai

Pills on the epithelial-mesenchymal transition (EMT) of trophoblasts and the embryo loss rate in embryos of mice with recurrent spontaneous abortion (RSA).

Methods

Ninety CBA/J female mice aged 6-8 weeks old were randomly divided into the normal group

the model group

the dydrogesterone group

the

Shoutai

Pills low-dose group

the

Shoutai

Pills medium-dose group and the

Shoutai

Pills high-dose group with 15 mice in each group. The normal pregnancy mouse was established with the mice in normal group × BALB/c male mice

and the RSA mice model was established with the mice in other groups × DBA/2 male mice. Female mice were administered doses 14 days before mating. The

Shoutai

Pills low-dose

medium-dose and high-dose groups were given 7.58

15.17

30.33 g/kg intragastric administration respectively

and dydrogesterone group was given dydrogesterone 0.003 g/kg intragastric administration

the normal group and the model group were given the same volume of distilled water intragastric administration. After continuous dosing to the 6th day of pregnancy

nine pregnant mice were selected from each group randomly and sacrificed under anesthesia

and then the embryos were harvested. Pathological changes of trophoblast cells at the maternal-fetal interface were observed by hematoxylin-eosin staining. Immunofluorescence staining was used to determine the protein expression levels

and real-time fluorescence quantitative PCR was used to determine the mRNA expression levels of EMT markers (E-cadherin

Vimentin) and β-catenin of embryonic trophoblasts in each group. The remaining pregnant mice in each group were administered doses until the 14th day of pregnancy

and after mice were sacrificed under anesthesia

embryo loss in each group was observed.

Results

Compared with the normal group

the protein and mRNA expression levels of the epithelial cell marker E-cadherin in the embryonic trophoblasts of the model group were increased (

0.05)

the expression levels of the mesenchymal cell marker Vimentin in the embryonic trophoblasts of the model group were decreased(

0.05)

the expression levels of β-catenin in the embryonic trophoblasts were decreased (

0.05)

and the embryo loss rate of the model group was increased (

0.05). Compared with the model group

the expression levels of E-cadherin in the embryonic trophoblasts were decreased (

0.05)

the expression levels of Vimentin in the embryonic trophoblasts were increased

and the expression levels of β-catenin in the embryonic trophoblasts were increased of each dose

Shoutai

Pills group and dydrogesterone group(

0.05)

all of which were most significant in the

Shoutai

Pills medium-dose and high-dose groups and the dydrogesterone group

the embryo loss rate in the

Shoutai

Pills low-dose group was decreased (

0.05)

and the embryo loss rate in the

Shoutai

Pills medium-dose and high-dose groups and the dydrogesterone group was decreased significantly (

0.05) .

Conclusion

Shoutai

Pills can reduce the rate of embryo loss in RSA mice

possibly by promoting EMT by activating β-catenin signaling molecules in trophoblasts and improving the migration and invasion ability of embryonic trophoblasts. These effects are conducive to the implantation of the embryo in the uterine cavity and play a role in protecting the fetus.

关键词

Keywords

references

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