Role of receptor for advanced glycation end-products and signalling events in advanced glycation end-product-induced monocyte chemoattractant protein-1 expression in differentiated mouse podocytes

作者：Leyi Gu, Shinji Hagiwara, Qiuling Fan, Mitsuo Tanimoto, Mami Kobata, Michifumi Yamashita, Tomohito Nishitani, Tomohito Gohda, Zhaohui Ni, Jiaqi Qian, Satoshi Horikoshi, Yasuhiko Tomino · 发表于：Nephrology Dialysis Transplantation · 年份：2005 · DOI：10.1093/ndt/gfi210 · 被引用次数：115 · 研究领域：Advanced Glycation End Products research、Chronic Kidney Disease and Diabetes、Renal Diseases and Glomerulopathies

BACKGROUND: Upregulation of local monocyte chemoattractant protein-1 (MCP-1) production is involved in glomerular damage through macrophage recruitment and activation in diabetic nephropathy. Treatment of db/db mice with soluble receptor for advanced glycation end-products (RAGE) prevented recruitment of macrophages to the glomeruli and reduced albuminuria, suggesting that binding of ligands and RAGE may be involved in MCP-1 expression. Therefore, we investigated the role of advanced glycation end-products (AGEs) in MCP-1 production by podocytes and signalling events after RAGE activation. METHODS: MCP-1 gene and protein expression were examined by using reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay in differentiated mouse podocytes. Dichlorofluorescein-sensitive intracellular reactive oxygen species (ROS) generation was measured by confocal microscopy. RAGE, phosphorylation of mitogen-activated protein kinases, nuclear factor (NF)-kappaB, c-Jun and Sp1 were studied using western blotting and immunocytochemistry. RESULTS: Both differentiated and undifferentiated podocytes expressed RAGE. MCP-1 was induced by AGEs and carboxymethyllysine (CML) in a time-dependent and dose-dependent manner in differentiated podocytes. Neutralizing antibody for RAGE suppressed AGE- and CML-induced MCP-1 production. AGEs and CML rapidly generated intracellular ROS in podocytes. Blocking of ROS by using N-acetyl-l-cysteine abolished CML and H(2)O(2)-induced ...