Human amnion mesenchymal stem cells promote endometrial repair via paracrine, preferentially than transdifferentiation

作者全名：Huang, Xiyue; Yang, Xiao; Huang, Jinglin; Wei, Ling; Mao, Yanhua; Li, Changjiang; Zhang, Yingfeng; Chen, Qiuhong; Wu, Shasha; Xie, Lele; Sun, Congcong; Zhang, Wenwen; Wang, Jia

作者地址：[Huang, Xiyue; Yang, Xiao; Huang, Jinglin; Wei, Ling; Mao, Yanhua; Li, Changjiang; Zhang, Yingfeng; Chen, Qiuhong; Wu, Shasha; Xie, Lele; Sun, Congcong; Zhang, Wenwen; Wang, Jia] Chongqing Med Univ, Univ Town Hosp, Dept Obstet & Gynecol, 55,Daxuecheng Middle Rd, Chongqing 401331, Peoples R China

通信作者：Sun, CC; Zhang, WW; Wang, J (通讯作者)，Chongqing Med Univ, Univ Town Hosp, Dept Obstet & Gynecol, 55,Daxuecheng Middle Rd, Chongqing 401331, Peoples R China.

来源：CELL COMMUNICATION AND SIGNALING

ESI学科分类：MOLECULAR BIOLOGY & GENETICS

WOS号：WOS:001236241100001

JCR分区：Q1

影响因子：8.2

年份：2024

卷号：22

期号：1

开始页：　

结束页：　

文献类型：Article

关键词：Intrauterine adhesion; Human amniotic mesenchymal stem cells; Paracrine, transdifferentiation; Endometrial fibrosis

摘要：Background Intrauterine adhesion (IUA) is one of the most severe causes of infertility in women of childbearing age with injured endometrium secondary to uterine performance. Stem cell therapy is effective in treating damaged endometrium. The current reports mainly focus on the therapeutic effects of stem cells through paracrine or transdifferentiation, respectively. This study investigates whether paracrine or transdifferentiation occurs preferentially in treating IUA.Methods Human amniotic mesenchymal stem cells (hAMSCs) and transformed human endometrial stromal cells (THESCs) induced by transforming growth factor beta (TGF-beta 1) were co-cultured in vitro. The mRNA and protein expression levels of Fibronectin (FN), Collagen I, Cytokeratin19 (CK19), E-cadherin (E-cad) and Vimentin were detected by Quantitative real-time polymerase chain reaction (qPCR), Western blotting (WB) and Immunohistochemical staining (IHC). The Sprague-Dawley (SD) rats were used to establish the IUA model. hAMSCs, hAMSCs-conditional medium (hAMSCs-CM), and GFP-labeled hAMSCs were injected into intrauterine, respectively. The fibrotic area of the endometrium was evaluated by Masson staining. The number of endometrium glands was detected by hematoxylin and eosin (H&E). GFP-labeled hAMSCs were traced by immunofluorescence (IF). hAMSCs, combined with PPCNg (hAMSCs/PPCNg), were injected into the vagina, which was compared with intrauterine injection.Results qPCR and WB revealed that FN and Collagen I levels in IUA-THESCs decreased significantly after co-culturing with hAMSCs. Moreover, CK19, E-cad, and Vimentin expressions in hAMSCs showed no significant difference after co-culture for 2 days. 6 days after co-culture, CK19, E-cad and Vimentin expressions in hAMSCs were significantly changed. Histological assays showed increased endometrial glands and a remarkable decrease in the fibrotic area in the hAMSCs and hAMSCs-CM groups. However, these changes were not statistically different between the two groups. In vivo, fluorescence imaging revealed that GFP-hAMSCs were localized in the endometrial stroma and gradually underwent apoptosis. The effect of hAMSCs by vaginal injection was comparable to that by intrauterine injection assessed by H&E staining, MASSON staining and IHC.Conclusions Our data demonstrated that hAMSCs promoted endometrial repair via paracrine, preferentially than transdifferentiation. IUA is the crucial cause of infertility in women of childbearing age, and no satisfactory treatment measures have been found in the clinic. hAMSCs can effectively treat intrauterine adhesions through paracrine and transdifferentiation mechanisms. This study confirmed in vitro and in vivo that amniotic mesenchymal stem cells preferentially inhibited endometrial fibrosis and promoted epithelial repair through paracrine, thus effectively treating intrauterine adhesions. The level of fibrosis marker proteins in IUA-THESCs decreased significantly after co-culturing with hAMSCs for 2 days in vitro. However, the level of epithelial marker proteins in hAMSCs increased significantly, requiring at least 6 days of co-culture. hAMSCs-CM had the same efficacy as hAMSCs in inhibiting fibrosis and promoting endometrial repair in IUA rats, supporting the idea that hAMSCs promoted endometrial remodeling through paracrine in vivo. In addition, GFP-labeled hAMSCs continuously colonized the endometrial stroma instead of the epithelium and gradually underwent apoptosis. These findings prove that hAMSCs ameliorate endometrial fibrosis of IUA via paracrine, preferentially than transdifferentiation, providing the latest insights into the precision treatment of IUA with hAMSCs and a theoretical basis for promoting the "cell-free therapy" of MSCs.

基金机构：Chongqing Municipal Science and Technology Bureau [cstc2021ycjh-bgzxm0014]; Natural Science Foundation of Chongqing [CSTB2023NSCQ-MSX0587]; Joint key project of Chongqing Health Commission and Science and Technology Bureau [2023ZDXM020]; Joint project of Chongqing Health Commission and Science and Technology Bureau [2024MSXM099]; Chongqing Education Commission [CYS21249, CYS22386]

基金资助正文：This work was supported by grants from Chongqing Municipal Science and Technology Bureau (cstc2021ycjh-bgzxm0014), Natural Science Foundation of Chongqing (CSTB2023NSCQ-MSX0587), Joint key project of Chongqing Health Commission and Science and Technology Bureau (2023ZDXM020), Joint project of Chongqing Health Commission and Science and Technology Bureau (2024MSXM099), Chongqing Education Commission (CYS21249, CYS22386).