IL-4-induced M2 macrophages inhibit fibrosis of endometrial stromal cells

Dan Feng; Yang Li; Hongyun Zheng; Ying Wang; Juexiao Deng; Tingting Liu; Wenxin Liao; Fujin Shen

doi:10.1016/j.repbio.2023.100852

IL-4-induced M2 macrophages inhibit fibrosis of endometrial stromal cells

Reprod Biol. 2024 Feb 13;24(2):100852. doi: 10.1016/j.repbio.2023.100852. Online ahead of print.

Authors

Dan Feng¹, Yang Li¹, Hongyun Zheng², Ying Wang¹, Juexiao Deng¹, Tingting Liu¹, Wenxin Liao¹, Fujin Shen³

Affiliations

¹ Department of Gynecology, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, PR China.
² Department of Clinical Laboratory, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, PR China.
³ Department of Gynecology, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, PR China. Electronic address: sfj296@163.com.

PMID: 38354656
DOI: 10.1016/j.repbio.2023.100852

Abstract

Background: Intrauterine adhesions (IUA) refers to endometrial fibrosis caused by irreversible damage of the endometrial basal layer. As the key regulators in tissue repair, regeneration, and fibrosis, macrophages play an essential role in endometrial regeneration and repair during the normal menstrual cycle. However, the mechanism of macrophages involved in IUA remains unclear.

Methods: In the late stages of proliferation, the endometrium was collected to make paraffin sections. HE and Masson staining were used to observing endometrial morphology and endometrial fibrosis. Immunohistochemistry and Western blotting were used to detect the expression level of fibrosis indexes COL1A1 and α-SMA. The macrophage infiltration was evaluated by immunohistochemistry for the expression levels of CD 206 and CD163. Next, we cultured the primary human endometrial stromal cells (HESCs), and then an IUA cell model was established with 10 ng/ml TGF-β1 for 72 h. THP 1 cells were differentiated by 100 ng/ml PMA into macrophages for 48 h, then macrophages were polarized to M2 macrophages by 20 ng/ml IL-4 for 24 h. The culture supernatants (M(IL-4) -S) of M2 macrophages were applied to the IUA cell model. The expression of fibrosis markers was then assessed using immunofluorescence and Western blotting.

Results: The results show that Patients with IUA have fewer endometrial glands and significantly increased fibrosis levels. Moreover, the infiltration of CD206-positive (M2) macrophages was significantly reduced in IUA patients, and negatively correlated with the expression of endometrial fibrosis indexes α-SMA and COL1A1. In addition, the primary HESCs treated with 10 ng/ml TGF-β1 for 72 h were found to have significantly increased levels of fibrosis indexes. Furthermore, supernatants from IL4-induced M2 macrophages inhibit the TGF-β1-induced fibrosis of HESCs.

Conclusions: M2 macrophages may negatively regulate the expression of COL1A1 and α-SMA, inhibiting the TGF-β1-induced fibrosis of HESCs. Our study suggests that targeting macrophage phenotypes and promoting the polarization of macrophages to M2 may become a novel strategy for the clinical treatment of IUA.

Keywords: Endometrium; Fibrosis; Human endometrial stromal cells; Intrauterine adhesions; Macrophages.