Effect of high glucose concentration on the synthesis of monocyte chemoattractant protein-1 in human peritoneal mesothelial cells: Involvement of protein kinase C
Effect of high glucose concentration on the synthesis of monocyte chemoattractant protein-1 in human peritoneal mesothelial cells: Involvement of protein kinase C
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DOI：
10.1159/000045941
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发表时间：
2001-04-01
期刊：
影响因子：
2.5
通讯作者：
Sitter, T
中科院分区：
文献类型：
--
作者：
Haslinger, B;Mandl-Weber, S;Sitter, T
人腹膜间皮细胞（HMC）通过其产生各种炎症介质的潜能而有助于腹膜中炎症过程的激活和控制。本研究旨在评估葡萄糖（大多数市售腹膜透析液中的渗透活性化合物）对培养的HMC中C-C趋化因子单核细胞趋化蛋白-1（MCP-1）合成的影响。用酶联免疫吸附法测定细胞培养上清中MCP-1的浓度，用北方印迹法检测MCP-1 mRNA的表达。HMC与葡萄糖（30-120 mM）孵育导致MCP-1蛋白分泌和mRNA表达的时间和浓度依赖性增加。24小时后，在用60 mM葡萄糖处理的HMC中，MCP-1合成从2.8 +/-0.46增加到4.2 +/-0.32 ng/10（5）细胞（n = 5，p < 0.05）。与此相反，渗透控制媒体含有代谢惰性单糖甘露醇或NaCl不影响MCP-1的生产。通过用佛波醇酯PMA（20 nM）激活蛋白激酶C（PKC）模拟高糖对HMC中MCP-1表达的刺激作用。将细胞与葡萄糖和特异性PKC抑制剂Ro 31-8220共孵育完全减弱了葡萄糖介导的MCP-1表达。总之，我们的研究结果表明，葡萄糖诱导MCP-1的合成PKC依赖性途径。由于渗透控制介质没有增加MCP-1的释放，这表明葡萄糖的作用主要与代谢有关，而与高渗性无关。这些数据可能部分解释了持续性非卧床腹膜透析患者透析流出液中MCP-1稳态水平升高的原因。版权所有2001 S. Karger AG.巴塞尔。
Human peritoneal mesothelial cells (HMC) contribute to the activation and control of inflammatory processes in the peritoneum by their potential to produce various inflammatory mediators. The present study was designed to assess the effect of glucose, the osmotic active compound in most commercially available peritoneal dialysis fluids, on the synthesis of the C-C chemokine monocyte chemoattractant protein-1 (MCP-1) in cultured HMC. The MCP-1 concentration in the cell supernatants was determined by enzyme-linked immunosorbent assay and the MCP-1 mRNA expression was examined using Northern blot analysis. Incubation of HMC with glucose (30-120 mM) resulted in a time- and concentration-dependent increase in MCP-1 protein secretion and mRNA expression. After 24 h the MCP-1 synthesis was increased from 2.8 +/- 0.46 to 4.2 +/- 0.32 ng/10(5) cells (n = 5, p < 0.05) in HMC treated with 60 mM glucose. In contrast, osmotic control media containing either the metabolically inert monosaccharide mannitol or NaCl did not influence MCP-1 production. The stimulating effect of high glucose on MCP-1 expression in HMC was mimicked by activation of protein kinase C (PKC) with the phorbol ester PMA (20 nM). Coincubation of the cells with glucose and the specific PKC inhibitor Ro 31-8220 completely blunted glucose-mediated MCP-1 expression. In summary, our results indicate that glucose induces MCP-1 synthesis by a PKC-dependent pathway. Since osmotic control media did not increase MCP-1 release, it is suggested that the effect of glucose is mainly related to metabolism and not to hyperosmolarity. These data may in part explain elevated steady-state levels of MCP-1 found in the dialysis effluent of continuous ambulatory peritoneal dialysis patients. Copyright 2001 S. Karger AG. Basel.