Chungsimyeonja-eum inhibits inflammatory responses in RAW 264.7 macrophages and HaCaT keratinocytes

Abstract

Background: Chungsimyeonja-eum (CSYJE) is an herbal prescription used in traditional Oriental medicine for treating cerebral infarction by reducing ischemic damage. However, the effects of CSYJE on inflammation have not been verified scientifically. Methods: Anti-inflammatory effects of CSYJE was investigated to dertermine the inhibitory effects of CSYJE against inflammation using RAW 264.7 mouse macrophages and HaCaT human keratinocytes. To measure the effects of CSYJE on inflammatory mediators and cytokines/chemokines, we used the following methods: cell viability assay, enzyme-linked immunosorbent assay (ELISA), western blotting, immunocytochemistry. RAW 264.7 cells were pretreated with CSYJE (250, 500, or 1000μg/mL) for 4h and treated with lipopolysaccharide (LPS) for additional 20h. HaCaT cells were stimulated with tumor necrosis factor alpha (TNF-aα) and interferon gamma (IFN-γ) (TI), and CSYJE (125, 250, or 500μg/mL) for 24h. Results: CSYJE suppressed the production of nitric oxide (NO, IC50 1000μg/mL), prostaglandin E2 (PGE2, IC50 = 12.1μg/mL), and interleukin (IL)-6 (IC50 = 248μg/mL) in LPS-stimulated RAW 264.7 cells. CSYJE suppressed the effects of TI on the production of thymus and activation-regulated chemokine (TARC, IC50 = 330.2μg/mL), macrophage-derived chemokine (MDC/CCL22, IC50 = 52.5μg/mL), regulated on activation, normal T-cell expressed and secreted (RANTES/CCL5, IC50 = 372.9μg/mL), and IL-8 (IC50 = 345.1μg/mL) in HaCaT cells. CSYJE inhibited TI-stimulated STAT1 phosphorylation in a dose-dependent manner and nuclear translocation at 500μg/mL in HaCaT cells. Conclusion: Our results suggest a possible therapeutic application of CSYJE for treating inflammatory diseases.