High glucose induces MCP-1 expression partly via tyrosine kinase-AP-1 pathway in peritoneal mesothelial cells
High glucose induces MCP-1 expression partly via tyrosine kinase-AP-1 pathway in peritoneal mesothelial cells
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DOI：
10.1046/j.1523-1755.2001.00770.x
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发表时间：
2001-07-01
影响因子：
19.6
通讯作者：
Park, JS
中科院分区：
文献类型：
--
作者：
Lee, SK;Kim, BS;Park, JS
背景腹膜透析液中的高糖与慢性非卧床腹膜透析（CAPD）患者腹膜纤维化的发病机制有关。然而.其机制尚不十分清楚。腹腔巨噬细胞似乎参与了腹膜纤维化的过程，单核细胞趋化蛋白-1（MCP-1）在单核细胞向腹腔募集中起关键作用。然而.高糖对人腹膜间皮细胞MCP-1表达及其信号转导通路的影响知之甚少。间皮细胞用葡萄糖（5 ~ 100 mmol/L）或甘露醇长期培养7 d。采用北方印迹法和酶联免疫吸附试验（ELISA）检测MCP-1 mRNA和蛋白的表达。用趋化实验测定高糖条件培养上清的趋化活性。采用凝胶电泳迁移率变动分析（EMSA）检测转录因子激活蛋白-1（AP-1）和核因子-κ B（NF-κ B）的表达。葡萄糖以时间和剂量依赖性方式诱导MCP-1 mRNA表达。细胞培养上清中MCP-1蛋白表达也明显增加。同等浓度的甘露醇无显著影响。高糖条件培养上清对单核细胞的趋化活性增加，可被抗MCP-1抗体中和。EMSA显示葡萄糖以时间和剂量依赖性方式增加AP-1结合活性。而不是NF-κ B。AP-Ⅰ抑制剂姜黄素呈剂量依赖性抑制高糖诱导的MCP-Ⅰ mRNA表达。酪氨酸激酶抑制剂如染料木素（12.5 ~ 50 μ mol/L）和除莠霉素A（0.1 ~ 1 μ mol/L）以剂量依赖方式抑制高糖诱导的MCP-Ⅰ mRNA表达。并抑制高糖诱导的AP-1结合活性。高糖部分通过酪氨酸激酶-AP-1途径诱导间皮细胞MCP-1表达。
Background. High glucose in peritoneal dialysis solutions has been implicated in the pathogenesis of peritoneal fibrosis in chronic ambulatory peritoneal dialysis (CAPD) patients. However. the mechanisms are not very clear. Peritoneal macrophages seem to participate in the process of peritoneal fibrosis and monocyte chemoattractant protein-1 (MCP-I) plays a key role in the recruitment of monocytes toward the peritoneal cavity. However. little is known about the effect of high glucose on MCP-1 expression and its signal transduction pathway in human peritoneal mesothelial cells.Methods. Mesothelial cells were cultured with glucose (5 to 100 mmol/L) or mannitol chronically for up to seven days. MCP-1 expression of mRNA and protein was measured by Northern blot analysis and enzyme-linked immunosorbent assay (ELISA). Chemotactic activity of high-glucose-conditioned culture supernatant was measured by chemotactic assay. To examine the roles of the transcription factors activator protein-1 (AP-1) and nuclear factor-KB (NF-kappaB), electrophoretic mobility shift assay (EMSA) was performed.Results. Glucose induced MCP-1 mRNA expression in a time-and dose-dependent manner. MCP-1 protein in cell culture supernant was also increased. Equivalent concentrations of mannitol had no significant effect. High-glucose-conditioned supernatant possessed an increased chemotactic activity for monocytes, which was neutralized by anti-MCP-1 antibody. EMSA revealed that glucose increased the AP-1 binding activity in a time- and dose-dependent manner. but not NF-kappaB. Curcumin, an inhibitor of AP-I, dose-dependently suppressed the induction of MCP-I mRNA by high glucose. Tyrosine kinase inhibitors such as genistein (12.5 to 50 mu mol/L) and herbimycin A (0.1 to I mu mol/L) inhibited the high-glucose-induced MCP-I mRNA expression in a dose-dependent manner. and also suppressed the high-glucose-induced AP-1 binding activity.Conclusions. High glucose induced mesothelial MCP-1 expression partly via the tyrosine kinase-AP-l pathway.