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Epithelial-mesenchymal Transition of Peritoneal Mesothelial Cells Is Enhanced by M2c Macrophage Polarization
Lifang Tian1, Qiaoling Yu1, Dan Liu1
1Department of Nephrology, Second Affiliated Hospital, Xi'an Jiaotong University, Xi'an, Shaanxi Province, P.R. China.
Background:
Peritoneal fibrosis (PF) can reduce the efficiency of peritoneal dialysis and eventually lead to ultrafiltration failure. Epithelial-mesenchymal transition (EMT) of peritoneal mesothelial cells (PMCs) is the start of PF. Macrophages are involved in the process. This study was to investigate the effect of macrophage polarization on EMT of PMCs.
Methods:
Monocyte-macrophage cells (THP-1) were treated to induce macrophage subsets (M1, M2a, M2c). The inducing was assessed by detecting protein and mRNA expression of cytokines using ELISA and RT-PCR. Subsequently, PMCs were co-cultured with M1, M2a and M2c, respectively, in Transwell chambers for 48 h and then expressions of E-cadherin and α-SMA were determined in PMCs. The PMCs that were not co-cultured with macrophages served as control PMCs. One-way ANOVA and SNK-q test were used to conduct statistics and P < .05 as significant.
Results:
Detection of the cytokines, including IL-6, IL-10, IL-12, TGF-β1, CCL17 and CXCL13, verified that the inducting of macrophage subtypes was successful. Compared to control, E-cadherin protein expression was significantly decreased and α-SMA protein expression increased in M1-treated PMCs (P < .05); M2a-treated PMCs had an increased gene expression of α-SMA (P < .05); E-cadherin protein and gene expression were decreased and α-SMA protein and gene expression increased significantly in M2c-treated PMCs (P < .05 or P < .01).
Conclusions:
EMT of PMCs is enhanced by M2c macrophage polarization; meanwhile, M1 and M2a polarization may have the effect to some extent, but not as definite as M2c.
Insights
Macrophage polarization significantly impacts peritoneal fibrosis by promoting epithelial-mesenchymal transition (EMT) in peritoneal mesothelial cells (PMCs). M2c macrophage polarization most strongly enhances EMT, suggesting a key role in peritoneal dialysis complications.
Area of Science:
- Nephrology
- Cell Biology
- Immunology
Background:
- Peritoneal fibrosis (PF) impairs peritoneal dialysis efficiency and can lead to ultrafiltration failure.
- Epithelial-mesenchymal transition (EMT) in peritoneal mesothelial cells (PMCs) initiates PF.
- Macrophages are implicated in the pathogenesis of PF.
Purpose of the Study:
- To investigate the influence of macrophage polarization on PMC EMT.
- To determine the specific roles of different macrophage subtypes (M1, M2a, M2c) in promoting EMT.
Main Methods:
- THP-1 cells were differentiated into M1, M2a, and M2c macrophage subsets.
- Cytokine expression (IL-6, IL-10, IL-12, TGF-β1, CCL17, CXCL13) confirmed successful polarization.
- PMCs were co-cultured with M1, M2a, or M2c macrophages.
- Expression of E-cadherin and α-SMA in PMCs was analyzed via ELISA and RT-PCR.
Main Results:
- M1 macrophage treatment decreased E-cadherin and increased α-SMA expression in PMCs.
- M2a macrophage treatment increased α-SMA gene expression in PMCs.
- M2c macrophage treatment significantly decreased E-cadherin and increased α-SMA expression in PMCs at both protein and gene levels.
Conclusions:
- M2c macrophage polarization significantly enhances EMT in PMCs.
- M1 and M2a macrophage polarization may contribute to EMT, but to a lesser extent than M2c.
- Targeting M2c macrophages could be a therapeutic strategy for peritoneal fibrosis.
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