Anti-oxidative and anti-inflammatory effect of Do-Ki-Tang methanol extract in mouse macrophage cells

Abstract

ABSTRACT Objective：The aim of this study was to determine whether methanol extract of Do-Ki-Tang (DKT) inhibit free radical generation and production of nitrite an index of NO, PG E 2 , iNOS, COX-2 and pro-inflammatory cytokines such as TNF-α, IL-1β, IL-6 and MCP-1 in lipopolysacch aride (LPS)-stimulated RAW 264.7 macrophages.Methods：Cytotoxic activity of extract on RAW 264.7 cells was measured using 5-(3-caroboxymeth -oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. The express ion level of inflammatory response-related proteins was confirmed by western blot. The production of proin flammatory cytokines was measured by ELISA.Results：Our results indicated that DKT scavenged DPPH radical and nitric oxide in vitro. Moreover, DKT significantly inhibited the LPS-induced NO, PGE 2 production and iNOS, COX-2 expression accompanied by an attenuation of TNF-α, IL-1β, IL-6 and MCP-1 formation in macrop hages. Furthermore, DKT treatment also blocked LPS-induced intracellular ROS production and the activation of NF-κB and MAPKs.Conclusion：Our data suggest that the anti-inflammatory effect of DKT is me diated through down-modulation of pro-inflammatory mediators and cytokines by blocking the signaling pathways of NF-κB and MAPKs. These inhibitory effects by DKT represent a potential therapeutic approach to the treatment of inflammatory diseases.Key words : Do-Ki-Tang (DKT), inflammation, macrophage