Alternative activation of macrophages in human peritoneum: implications for peritoneal fibrosis
Alternative activation of macrophages in human peritoneum: implications for peritoneal fibrosis
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DOI：
10.1093/ndt/gfq771
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发表时间：
2011-09-01
影响因子：
6.1
通讯作者：
Auxiliadora Bajo, Maria
中科院分区：
文献类型：
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作者：
Bellon, Teresa;Martinez, Virginia;Auxiliadora Bajo, Maria
背景取决于细胞因子微环境，巨噬细胞（M phi）可以采用促炎（M1）或促纤维化（M2）表型，其特征在于细胞表面蛋白如CD 206和CD 163以及可溶性因子如CC趋化因子配体18（CCL 18）的表达。已在不同器官环境中观察到M phi在纤维化中的关键作用。我们在腹膜透析的人类模型中研究了M phi人群，其中透析液和复发性腹膜炎引起的持续应激代表了腹膜功能障碍的风险，反映为超滤失败（UFF）和腹膜纤维化。我们使用流式细胞术和定量逆转录-聚合酶链反应来分析腹膜流出物M phi的表型，并测试它们刺激人成纤维细胞增殖的能力。比较非感染患者与活动性腹膜炎患者的M phi。通过酶联免疫吸附试验（ELISA）评价用过的透析液和细胞培养上清液中细胞因子的产生。流式细胞术分析发现腹膜流出液中存在CD 206（+）和CD 163（+）M2，腹膜炎期间CD 163（+）细胞的频率增加（P = 0.003）。腹腔巨噬细胞中TGF β 1、MMP 9和CCL 18信使RNA（mRNA）水平（pM phi）与体外分化的M2细胞中发现的水平相似。pM phi刺激成纤维细胞增殖的能力与CCL 18 mRNA水平相关（r = 0.924，P = 0.016）。通过细胞离心涂片样品的免疫染色和ELISA证实pM phi产生CCL 18。此外，CCL 18流出液浓度与腹膜功能下降相关，其评价指标为透析液与血浆肌酐比值（r = 0.724，P < 0.0001），且在UFF患者（P = 0.0025）和后来发生硬化性腹膜炎的患者（P = 0.024）中显著更高。M2可能通过刺激成纤维细胞生长和CCL 18产生参与人腹膜纤维化，因为高浓度的CCL 18与腹膜功能缺陷和纤维化相关。
Background. Depending on the cytokine microenvironment, macrophages (M phi) can adopt a proinflammatory (M1) or a profibrotic (M2) phenotype characterized by the expression of cell surface proteins such as CD206 and CD163 and soluble factors such as CC chemokine ligand 18 (CCL18). A key role for M phi in fibrosis has been observed in diverse organ settings. We studied the M phi population in a human model of peritoneal dialysis in which continuous stress due to dialysis fluids and recurrent peritonitis represent a risk for peritoneal membrane dys-function reflected as ultrafiltration failure (UFF) and peritoneal fibrosis.Methods. We used flow cytometry and quantitative reverse transcription-polymerase chain reaction to analyse the phenotype of peritoneal effluent M phi and tested their ability to stimulate the proliferation of human fibroblasts. M phi from non-infected patients were compared with those from patients with active peritonitis. Cytokine production was evaluated by enzyme-linked immunosorbent assay (ELISA) in spent dialysates and cell culture supernatants.Results. CD206(+) and CD163(+) M2 were found within peritoneal effluents by flow cytometry analysis, with increased frequencies of CD163(+) cells during peritonitis (P = 0.003). TGFB1, MMP9 and CCL18 messenger RNA (mRNA) levels in peritoneal macrophages (pM phi) were similar to those found in M2 cells differentiated in vitro. The ability of pM phi to stimulate fibroblast proliferation correlated with CCL18 mRNA levels (r = 0.924, P = 0.016). CCL18 production by pM phi was confirmed by immunostaining of cytospin samples and ELISA. Moreover, CCL18 effluent concentrations correlated with decreased peritoneal function, which was evaluated as dialysate to plasma ratio of creatinine (r = 0.724, P < 0.0001), and were significantly higher in patients with UFF (P = 0.0025) and in those who later developed sclerosing peritonitis (P = 0.024).Conclusions. M2 may participate in human peritoneal fibrosis through the stimulation of fibroblast cell growth and CCL18 production as high concentrations of CCL18 are associated with functional deficiency and fibrosis of the peritoneal membrane.