Nafamostat mesilate suppresses NF-κB activation and NO overproduction in LPS-treated macrophages

• DOI: 10.1016/s1567-5769(03)00146-2
• Publication Date: 2003-06-30T13:57:08Z
• Authors: Soichi Noguchi; Mikiya Nakatsuka; Hideki Konishi; Yasuhiko Kamada; Chebib Chekir; Takafumi Kudo
• Keywords: Cell Nucleus; Lipopolysaccharides; 0301 basic medicine; Transcription, Genetic; Macrophages; Anti-Inflammatory Agents, Non-Steroidal; NF-kappa B; Nitric Oxide Synthase Type II; Electrophoretic Mobility Shift Assay; Nitric Oxide; Guanidines; Benzamidines; Cell Line; 3. Good health; Mice; 03 medical and health sciences; NG-Nitroarginine Methyl Ester; Depression, Chemical; Enzyme Induction; Animals; I-kappa B Proteins; Enzyme Inhibitors; Nitric Oxide Synthase

Nafamostat mesilate (NM), a clinically used serine protease inhibitor, suppressed the overproduction of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS) in RAW264.7 murine macrophages treated with lipopolysaccharide (LPS, 100 ng/ml); however, it had little effect on endothelial NOS (eNOS) in human umbilical vein endothelial cells (HUVEC). Electrophoretic mobility shift assay (EMSA) revealed that LPS activated nuclear factor-kappaB (NF-kappaB) in RAW264.7 cells and that this activation was suppressed by nafamostat mesilate. Western blotting showed that nafamostat mesilate suppressed the phosphorylation and degradation of inhibitor kappaB-alpha (IkappaB-alpha), which holds NF-kappaB in the cytoplasm in an inactivated state. Our observations suggest that nafamostat mesilate is a candidate agent for various diseases such as ischemia-reperfusion, graft rejection, inflammatory diseases, and autoimmune diseases, in which iNOS and/or NF-kappaB are upregulated.