Apoptotic bodies of bone marrow mesenchymal stem cells inhibit endometrial stromal cell fibrosis by mediating the Wnt/β-catenin signaling pathway

Zhenghua Xiong; Yaru Ma; Jia He; Qin Li; Liu Liu; Chunli Yang; Jia Chen; Yi Shen; Xuesong Han

doi:10.1016/j.heliyon.2023.e20716

Apoptotic bodies of bone marrow mesenchymal stem cells inhibit endometrial stromal cell fibrosis by mediating the Wnt/β-catenin signaling pathway

Heliyon. 2023 Oct 13;9(11):e20716. doi: 10.1016/j.heliyon.2023.e20716. eCollection 2023 Nov.

Authors

Zhenghua Xiong^{1

2}, Yaru Ma³, Jia He⁴, Qin Li², Liu Liu⁴, Chunli Yang⁵, Jia Chen², Yi Shen², Xuesong Han^{1

2}

Affiliations

¹ Department of Gynecology, First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan, China.
² Department of Gynecology, Yan'an Hospital Affiliated to Kunming Medical University, Kunming, Yunnan, China.
³ Department of Gynecology, Women and Children's Hospital Affiliated to Qingdao University, Qingdao, Shandong, China.
⁴ Department of Plastic Surgery, Affiliated Calmette Hospital of Kunming Medical University, Kunming, Yunnan, China.
⁵ Department of Gynecology, Baoshan People's Hospital, Baoshan, Yunnan, China.

Abstract

Background: Intrauterine adhesions (IUAs) are a common illness of the uterine cavity. Endometrial fibrosis is the main pathological feature. In addition to a high recurrence rate, patients with severe IUAs have a low pregnancy rate. However, there are few effective treatments for IUAs. This study aims to confirm the influence of apoptotic bodies of bone marrow mesenchymal stem cells (BMSCs) on endometrial stromal cell fibrosis by mediating the Wnt/β-catenin signaling pathway and to provide new insight for the clinical treatment of IUAs.

Methods: Human endometrial stromal cells (HESCs) were used to establish an IUA cell model by treatment with TGF-β1, and a rat IUA model was established by the double injury method. Apoptosis of BMSCs was detected by TUNEL assays, and cell morphology was observed by the CM-DiI tracer. The morphology of apoptotic vacuoles and apoptotic bodies (ABs) was detected by TEM. We used Western blotting to detect the expression of histone H3.3, histone H2B, C3b, cyclin D1, C1QC, α-SMA, COL1A1, COL5A2, FN, CTGF, Wnt2b, c-MYC, CK-18 and VIM. The expression levels of α-SMA, COL1A1, COL5A2, FN and CTGF were detected by RT‒qPCR. The expression levels of α-SMA, COL1A1, FN and CTGF were detected by immunofluorescence. Immunohistochemistry was used to detect the expression of TGF-β, CK-18 and VIM. Flow cytometry, cell scratch assays, CCK-8 assays, and H & E and Masson staining were used to detect the cell cycle, cell migration, cell proliferation, and endometrial pathology, respectively.

Results: We found that ultraviolet light (UV) irradiation induced apoptosis of BMSCs and increased the production of ABs. TGF-β1 treatment can induce HESCs to form extracellular matrix (ECM), and aggravate cell fibrosis, and adding ABs or FH535, an inhibitor of the Wnt/β-catenin signaling pathway, can inhibit TGF-β1-induced HESC fibrosis. However, the inhibitory effect of ABs on TGF-β1-induced fibrosis of HESCs was attenuated by the addition of LiCl. In the Wnt/β-catenin signaling pathway, LiCl is an activator after coculture with TGF-β1. In vivo, IUA-induced narrowing of the uterine cavity was accompanied by intrauterine adhesions, increased deposition of collagen fibers, upregulation of TGF-β1, VIM, α-SMA, COL1A1 and COL5A2, and downregulation of CK-18. These changes in expression were reversed after treatment with ABs or FH535. When ABs and LiCl were added at the same time, the inhibitory effect of ABs on IUA fibrosis was weakened.

Conclusion: BMSC-derived ABs inhibit the fibrosis of HESCs by inhibiting the Wnt/β-catenin signaling pathway. These results provide a new direction for the clinical treatment of IUAs.

Keywords: Apoptotic body; Bone marrow mesenchymal stem cells; Endometrial stromal cells; Fibrosis; Wnt/β-catenin signaling pathway.