Investigation of Anti-inflammatory and Anti-oxidative Activities of Lonicerae Flos, <i>Citri</i> Pericarpium and <i>Violae</i> Herba Complex (LCVC)

Abstract

Objectives: The anti-inflammatory and anti-oxidative activities of LCVC (<i>Lonicerae</i> Flos, <i>Citri</i> Pericarpium and <i>Violae</i> Herba Complex) have not been fully elucidated. The purpose of this study was to investigate the mechanisms underlying these effects in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages.Methods: The evaluation of the anti-oxidative activity of LCVC was completed via DPPH and ABTS radical scavenging capacity, FRAP assay, measurement of polyphenol and flavonoid, assessment of ROS and NO levels in LPS-induced RAW 264.7 cells. The anti-inflammatory activity was defined by measuring the production of biomarkers (PGE2, IL-1B, IL-6 and TNF-<i>α</i>), proteins (ERK, JNK, P38, Nrf2, Keap1, HO-1 and NQO1) and expressions of genes (iNOS, COX-2, IL-1<i>β</i>, IL-6, TNF-<i>α</i>, Nrf2, Keap1, HO-1 and NQO1) in LPS-induced RAW 264.7 cells.Results: LCVC have polyphenol and flavonoid contents. The results of DPPH and ABTS free radical scavenging capacity and FRAP assay showed that the anti-oxidative activity was increased. Production of ROS, NO, IL-6, TNF-<i>α</i>, mRNA expressions of IL-1<i>β</i>, IL-6, TNF-<i>α</i>, Keap1, iNOS and COX-2 were decreased, and NQO1, Nrf2, and HO-1 were increased. In protein expression, JNK and Keap1 were decreased, NQO1, Nrf2 and HO-1 were increased, and no relationships were observed with the ERK and P38 by LCVC.Conclusions: These results suggest that LCVC may offer protective effects against LPS-induced inflammatory and oxidative responses through attenuating Nrf2/HO-1 pathway and MAPKs pathway. Therefore, we propose that LCVC has anti-inflammatory and anti-oxidative activities that have therapeutic potential in the treatment of inflammatory and oxidative disorders caused by the over-activation of macrophages.