Interaction of M2 macrophages and endometrial cells induces downregulation of GRIM-19 in endometria of adenomyosis

Wang B, Bingyu Wang, Yang Yang, Yang Y, Deng X, Xiaohui Deng, Ban Y, Yanli Ban, Chao Lan, Chao L, Lan Chao
Reproductive biomedicine online · 2020 · vol. 41(5) , pp. 790–800 · doi:10.1016/j.rbmo.2020.04.022 · PMID:32896475 · W3024114123
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M2 macrophages interacting with endometrial cells were observed to downregulate GRIM-19 expression in the endometria of patients with adenomyosis.

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Abstract

Research questionDoes the aggregation of M2 macrophages affect the expression of gene associated with retinoid-interferon-induced mortality 19 (GRIM-19) in adenomyosis?DesignEndometrial tissues were collected from patients with (n = 15) and without (n = 15) adenomyosis. Tissues were analysed for GRIM-19 and toll-like receptor 4 (TLR4) expression by immunohistochemistry and western blotting. Apoptosis was analysed by TdT (terminal deoxynucleotidyl transferase)-mediated dUDP nick-end labelling (TUNEL) assay. Human endometrial stromal cells (HESC) were transfected with GRIM-19 small interfering RNA (SiRNA) to knockdown GRIM-19 expression. The HESC were co-cultured with M2 macrophages to detect the influence of M2 macrophages in HESC cells. Analyses included GRIM-19, caspase-3 and TLR4 expression by western blotting, and GRIM-19 and TLR4 by quantitative real-time polymerase chain reaction. Apoptosis was measured by flow cytometry and TUNEL assay. Cell proliferation (Cell Counting Kit-8 assay) and migration assays were carried out.ResultsThe expression of GRIM-19 was significantly lower in adenomyosis lesions compared with controls (P < 0.001). Deficiency of GRIM-19 induced by siRNA decreased apoptosis and increased proliferation and migration in HESC. A significant decrease in GRIM-19 expression occurred in HESC after co-culture with M2 macrophages (P = 0.018). After co-culture with M2 macrophage, apoptosis decreased and proliferation and cell invasion in HESC increased. Protein (P = 0.006) and mRNA (P = 0.013) expression of TLR4 in HESC also reduced after this co-culture. Up-regulation of GRIM-19 occurred in HESC treated with the activator TLR4 (P = 0.016). Up-regulation of GRIM-19 was significantly reversed in cells treated with the TLR4 inhibitor (P = 0.011).ConclusionM2 macrophages may be involved in regulating the expression of GRIM-19 partly through the TLR4 signalling axis in adenomyosis.

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Condition tags

adenomyosis

MeSH descriptors

Adenomyosis Apoptosis Regulatory Proteins Endometrium Macrophages NADH, NADPH Oxidoreductases Toll-Like Receptor 4 Adenomyosis Adenomyosis Adolescent Adult Apoptosis Apoptosis Apoptosis Regulatory Proteins Apoptosis Regulatory Proteins Caspase 3 Caspase 3 Cell Proliferation Cell Proliferation Down-Regulation Endometrium

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