Role of Notch signaling pathway in treatment of intrauterine adhesions through human amniotic mesenchymal stem cells transplantation

Jie YU; Wenwen ZHANG; Jiayue HUANG; Yating GOU; Yanhua MAO; Yingfeng ZHANG; Congcong SUN; Benyuan WU; Changjiang LI; Nizhou LIU; Tingting WANG; Jiren HUANG; Jia WANG

doi:10.16016/j.1000-5404.202102021

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Role of Notch signaling pathway in treatment of intrauterine adhesions through human amniotic mesenchymal stem cells transplantation

Basic Medicine | 更新时间：2026-01-09

- Role of Notch signaling pathway in treatment of intrauterine adhesions through human amniotic mesenchymal stem cells transplantation
- Journal of Third Military Medical University Vol. 43, Issue 14, Pages: 1346-1357(2021)
- 作者机构：
  
  401331 重庆，重庆医科大学附属大学城医院妇产科
- 作者简介：
- 基金信息：
  
  the Basic Research and Frontier Exploration Project of Yuzhong District, Chongqing(20190122);the Postgraduate Scientific Research and Innovation Project of Chongqing(CYS19195)
- DOI：10.16016/j.1000-5404.202102021
  CLC： R329.24;R394.2;R711.74
- Received：03 February 2021，
  
  Revised：2021-03-31，
  
  Published：30 July 2021
- 稿件说明：
移动端阅览
Jie YU, Wenwen ZHANG, Jiayue HUANG, et al. Role of Notch signaling pathway in treatment of intrauterine adhesions through human amniotic mesenchymal stem cells transplantation[J]. Journal of Third Military Medical University, 2021, 43(14): 1346-1357.
DOI：

Jie YU, Wenwen ZHANG, Jiayue HUANG, et al. Role of Notch signaling pathway in treatment of intrauterine adhesions through human amniotic mesenchymal stem cells transplantation[J]. Journal of Third Military Medical University, 2021, 43(14): 1346-1357. DOI： 10.16016/j.1000-5404.202102021.

摘要

目的

探讨Notch信号通路在人羊膜间充质干细胞(human amniotic mesenchymal stem cells，hAMSCs)移植治疗宫腔粘连(intrauterine adhesions，IUA)过程中的作用。

方法

体外实验: 将细胞分为3组，即hAMSCs空白对照组、AdRFP腺病毒转染hAMSCs组、AdR-dnNotch1腺病毒转染hAMSCs组；RT-PCR检测各组中Notch1 mRNA水平，Western blot检测NICD的蛋白表达改变。体内实验: 40只Sprague-Dawley (SD)雌性大鼠分为4组，即假手术组、IUA组、hAMSCs移植治疗组、腺病毒(AdR-dnNotch1)转染hAMSCs后移植治疗组；取各组子宫组织，HE染色检测子宫内膜腺体数目和子宫内膜厚度变化，Masson染色检测纤维化程度变化，免疫组化检测Vimentin、Notch1、NICD、Hes1蛋白表达变化，Western blot检测各组子宫组织中NICD的蛋白表达量，RT-PCR检测各组中Hes1 mRNA水平。通过雌雄合笼评估各组SD大鼠生育力的变化。

结果

体外实验: Notch1显性负性突变腺病毒(AdR-dnNotch1)可成功抑制Notch信号通路。体内实验: HE、Masson染色结果显示，与IUA组比较，hAMSCs移植组可使腺体数目增多，子宫内膜厚度变薄，纤维化面积比减少(

0.05)，而与IUA组比较，AdR-dnNotch1转染hAMSCs组变化不明显。免疫组化结果显示，与假手术组比较，IUA组中Vimentin表达升高，Notch1、NICD、Hes1表达降低(

0.05)。与IUA组比较，hAMSCs移植组中Vimentin表达下降，Notch1、NICD、Hes1表达增加(

0.05)，AdR-dnNotch1腺病毒转染组Vimentin、Notch1、NICD、Hes1蛋白表达变化差异无统计学意义；Western blot、RT-PCR结果显示，Notch信号通路在hAMSCs移植治疗组中处于激活状态；雌雄合笼结果显示，与IUA组比较，hAMSCs移植组胚胎数量明显增加(

0.05)，而AdR-dnNotch1转染hAMSCs移植组胚胎数量无明显增加。

结论

Notch信号通路参与调控hAMSCs生物学功能，抑制Notch信号通路后，hAMSCs移植治疗宫腔粘连的作用降低。

Abstract

Objective

To explore the role of Notch signaling pathway in the transplantation of human amniotic mesenchymal stem cells (hAMSCs) for treatment of intrauterine adhesions (IUA).

Methods

In vitro

experiments: hAMSCs were isolated from human amnion obtained from term cesarean section

and then induced and identified. Then the cells were divided into 3 groups: blank control group (hAMSCs only)

AdRFP group (Ad-RFP adenovirus+hAMSCs)

and AdR-dnNotch1+hAMSCs group (a recombinant adenovirus expressing the dominant-negative mutant of Notch1+hAMSCs)

respectively. Then the mRNA level of Notch1 and the protein level of Notch intracellular domain (NICD) were detected by RT-PCR and

Western blotting

respectively.

In vivo

experiments: 40 Sprague-Dawley (SD) female rats were randomized into Sham group

IUA group

hAMSCs group (IUA animals treated with hAMSCs transplantation)

AdR-dnNotch1+hAMSCs group (IUA animals receiving transplantation of AdR-dnNotch1-transfected hAMSCs) (

=10). The number of endometrial glands and the thickness of endometrium in each group were observed by HE staining

and the changes of fibrosis were studied by Masson staining. Immunohistochemical assay was adopted to detect the protein expression of Vimentin

Notch1

NICD

and Hes1 in each group

and that of NICD was further tested by Western blotting. In addition

RT-PCR was used to determine the mRNA level of Hes1. Finally

the changes of fertility in each group were evaluated by co-cage of male and female rats.

Results

AdR-dnNotch1 successfully inhibited Notch signaling pathway

in vitro. In vivo

experiments: HE and Masson staining showed that when compared with IUA group

the number of glands was increased

and the thickness of endometrium and the ratio of fibrosis area were decreased in hAMSCs group (

0.05)

while no significant changes were found in the AdR-dnNotch1+hAMSCs group (

0.05); Immunohistochemical assay indicated that the level of Vimentin was higher and those of Notch1

NICD and Hes1 were lower in the IUA group than the Sham group (

0.05). But the above changes were reversed in the hAMSCs group as compared with the IUA group (

0.05)

and there were no significant differences in the AdR-dnNotch1+hAMSCs group (

0.05); Western blotting and RT-PCR results suggested the Notch signaling pathway was activated in hAMSCs group; The co-cage experiment showed that the number of embryos was increased significantly in hAMSCs group (

0.05)

though without noticeable changes in AdR-dnNotch1+hAMSCs group (

0.05).

Conclusion

Notch signaling pathway is involved in the regulation of biological functions of hAMSCs

and inhibition of Notch signaling pathway may reduce the effect of hAMSCs transplantation in the treatment of IUA.

关键词

Keywords

references

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