Department of Nephrology, Second Affiliated Hospital, Xi'an Jiaotong University, Xi'an, Shaanxi Province, P.R. China.
Medical Laboratory, Second Affiliated Hospital, Xi'an Jiaotong University, Xi'an, Shaanxi Province, P.R. China.
Immunol Invest. 2022 Feb;51(2):301-315. doi: 10.1080/08820139.2020.1828911. Epub 2021 Sep 7.
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.
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 < .05 as significant.
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 ( < .05); M2a-treated PMCs had an increased gene expression of α-SMA ( < .05); E-cadherin protein and gene expression were decreased and α-SMA protein and gene expression increased significantly in M2c-treated PMCs ( < .05 or < .01).
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.
腹膜纤维化(PF)会降低腹膜透析效率,最终导致超滤失败。腹膜间皮细胞(PMCs)的上皮-间充质转化(EMT)是 PF 的开始。巨噬细胞参与这一过程。本研究旨在探讨巨噬细胞极化对 PMCs EMT 的影响。
单核-巨噬细胞细胞(THP-1)经诱导分化为巨噬细胞亚群(M1、M2a、M2c),通过酶联免疫吸附试验(ELISA)和逆转录聚合酶链反应(RT-PCR)检测细胞因子蛋白和 mRNA 的表达来评估诱导效果。随后,将 PMCs 分别与 M1、M2a 和 M2c 共培养于 Transwell 小室中 48 h,检测 PMCs 中 E-钙黏蛋白和α-SMA 的表达。未与巨噬细胞共培养的 PMCs 作为对照 PMCs。采用单因素方差分析和 SNK-q 检验进行统计学分析, < 0.05 为差异有统计学意义。
检测细胞因子(包括 IL-6、IL-10、IL-12、TGF-β1、CCL17 和 CXCL13)表明,巨噬细胞亚型的诱导成功。与对照组相比,M1 处理的 PMCs 中 E-钙黏蛋白蛋白表达明显降低,α-SMA 蛋白表达增加( < 0.05);M2a 处理的 PMCs 中α-SMA 基因表达增加( < 0.05);M2c 处理的 PMCs 中 E-钙黏蛋白蛋白和基因表达降低,α-SMA 蛋白和基因表达明显增加( < 0.05 或 < 0.01)。
M2c 极化的巨噬细胞增强了 PMCs 的 EMT;同时,M1 和 M2a 极化在某种程度上可能有一定作用,但不如 M2c 确定。