Effect of Salicylate on the Monocyte Chemoattractant Protein-1 Expression and Intracellular Reactive Oxygen Species Formation in Human Mesangial Cells.
- Author:
Shi Jung CHUNG
1
;
Choung Soo KIM
;
Jae Won CHANG
;
Soon Bae KIM
;
Sang Koo LEE
;
Jung Sik PARK
Author Information
1. Department of Internal Medicine, Asan Medical Center, College of Medicine, University of Ulsan, Seoul, Korea. sklee2@amc.seoul.kr
- Publication Type:Original Article
- Keywords:
Mesangial cell;
Salicylate;
Monocyte chemoattractant protein-1;
Nuclear factor-kappaB;
Reactive oxygen species
- MeSH:
Aspirin;
Blotting, Northern;
Blotting, Western;
Chemokine CCL2*;
Electrophoretic Mobility Shift Assay;
Enzyme-Linked Immunosorbent Assay;
Flow Cytometry;
Hand;
Humans*;
Inflammation;
Interleukin-1beta;
Lysophosphatidylcholines;
Mesangial Cells*;
Monocytes*;
NF-kappa B;
Reactive Oxygen Species*;
RNA, Messenger;
Tumor Necrosis Factor-alpha
- From:Korean Journal of Nephrology
2003;22(3):261-272
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: Monocyte chemoattractant protein-1 (MCP-1) and reactive oxygen species (ROS) play an important role during glomerular inflammation. We investigated the effect of aspirin metabolite, salicylate on the pro-inflammatory cytokine-induced MCP-1 expression and lysophosphatidylcholine -induced intracellular ROS formation in human mesangial cells. METHODS: Cells were pretreated with salicylate, and then stimulated with tumor necrosis factor-alpha (TNF-alpha) or interleukin-1beta (IL-1beta). The expression of MCP-1 mRNA and MCP-1 protein were measured by Northern blot analysis and enzyme-linked immunosorbent assay, respectively. Nuclear factor-kappa B (NF-kappaB) activity was measured by electrophoretic mobility shift assay. Degradation of Ikappa B-alpha was assessed by Western blot analysis. Intracellular ROS production was monitored by flow cytometry using 2'7'-dichlorofluorescin diacetate. RESULTS: Salicylate inhibited the TNF-alpha- or IL-1beta- induced MCP-1 mRNA expression in a dose dependent manner (1-20 mM) and also suppressed the MCP-1 protein expression. Its effect was not attributable to de novo synthesis of intermediary proteins. Salicylate inhibited the TNF-alpha- or IL-1beta-induced NF-kappa B binding activity and also suppressed the TNF-alpha-induced Ikappa B-alpha degradation. Low concentration of salicylate (0.01-1 mM) suppressed the lysophosphatidylcholine-induced ROS formation. CONCLUSION: Milimolar concentration of salicylate inhibited the MCP-1 expression at least in part, via suppression of NF-kappaB by reducing the degradation of Ikappa B-alpha. On the other hand, lower concentration of salicylate could suppress the lysophosphatidylcholine-induced intracellular ROS formation.