高糖部分通过酪氨酸激酶-AP-1途径诱导腹膜间皮细胞中MCP-1的表达。

High glucose induces MCP-1 expression partly via tyrosine kinase-AP-1 pathway in peritoneal mesothelial cells.

作者信息

Lee S K, Kim B S, Yang W S, Kim S B, Park S K, Park J S

机构信息

Department of Internal Medicine, and Asan Institute for Life Sciences, University of Ulsan, College of Medicine, Seoul, Korea.

出版信息

Kidney Int. 2001 Jul;60(1):55-64. doi: 10.1046/j.1523-1755.2001.00770.x.

DOI:10.1046/j.1523-1755.2001.00770.x

PMID:11422736

Abstract

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-1) 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-kappaB (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-1, dose-dependently suppressed the induction of MCP-1 mRNA by high glucose. Tyrosine kinase inhibitors such as genistein (12.5 to 50 micromol/L) and herbimycin A (0.1 to 1 micromol/L) inhibited the high-glucose-induced MCP-1 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-1 pathway.

摘要

背景

腹膜透析液中的高糖已被认为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，其机制尚不完全清楚。腹膜巨噬细胞似乎参与了腹膜纤维化过程，单核细胞趋化蛋白-1（MCP-1）在单核细胞向腹腔的募集中起关键作用。然而，关于高糖对人腹膜间皮细胞中MCP-1表达及其信号转导途径的影响知之甚少。

方法

将间皮细胞分别用葡萄糖（5至100 mmol/L）或甘露醇长期培养长达7天。通过Northern印迹分析和酶联免疫吸附测定（ELISA）检测MCP-1 mRNA和蛋白的表达。通过趋化试验测定高糖条件培养上清液的趋化活性。为了研究转录因子激活蛋白-1（AP-1）和核因子-κB（NF-κB）的作用，进行了电泳迁移率变动分析（EMSA）。

结果

葡萄糖以时间和剂量依赖性方式诱导MCP-1 mRNA表达。细胞培养上清液中的MCP-1蛋白也增加。同等浓度的甘露醇无显著影响。高糖条件培养上清液对单核细胞具有增强的趋化活性，抗MCP-1抗体可使其中和。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，但不增加NF-κB结合活性。姜黄素是一种AP-1抑制剂，可剂量依赖性地抑制高糖诱导的MCP-1 mRNA表达。酪氨酸激酶抑制剂如染料木黄酮（12.5至50 μmol/L）和除莠霉素A（0.1至1 μmol/L）以剂量依赖性方式抑制高糖诱导的MCP-1 mRNA表达，并抑制高糖诱导的AP-1结合活性。