Anti-oxidative and anti-inflammatory experiments of Talmyung-san in RAW264.7 cells

Objectives : The aim of this study was identification of the anti-oxidative and anti-inflammatory effects of Talmyung-san (TMS) in mouse macrophage, RAW264.7 cells. Methods : To identify the anti-oxidative effect of TMS, scavenging activities of DPPH radical, nitric oxide and peroxynitrite were measured in vitro. In RAW264.7 cells, DCFH-DA assay was conducted to examine the inhibitory effect of TMS on ROS production in response to lipopolysaccharide. And the productions of nitric oxide (NO), PGE₂ and pro-inflammatory cytokines were measured. The levels of COX-2, iNOS, nuclear NF- κB p65 expression and phosphorylation of IκB-α in cytosol were detected by western blotting analyses. Results : TMS couldn't scavenged DPPH radical, but nitric oxide and peroxynitrite were decreased. TMS decreased intracellular ROS, NO, and IL- 1β production effectively. However, TMS inhibited PGE₂ levels only in high concentration ( 300μg/m ) and TMS failed to suppress the production of IL-6 and TNF- α . Results from immunoblot analyses revealed that TMS decreased activation of COX-2, iNOS, phosphorylation of IκB-α and nuclear translocation of p65. Conclusions : TMS has anti-RNS and anti-inflammatory effects via NF- κB pathway and more intensive studies will be required to evaluate therapeutic potential of TMS.