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OBJECTIVE: This study aimed to evaluate whether Hwangryunhaedoktang(HHT), a herbal compound, has an inhibitory effect on lipopolysaccharide(LPS)-induced inflammation in RAW264.7 macrophages.METHODS: The effects of HHT were evaluated by confirming nitric oxide(NO) production and expression of inducible NO synthase(iN OS) and mitogen-activated protein kinases(MAPKs) in LPS-stimulated RAW264.7 macrophages via the Griess assay, Western blotting, and real-time reverse transcription quantitative polymerase chain reaction. Western blot analyses and luciferase assays were used to evaluate whether HHT has an effect on the phosphorylation and translocation of nuclear factor-κB(NF-κB). The secretion and expression of tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) were determined via enzyme-linked immunosorbent assay and Western blot analyses.RESULTS: HHT suppressed LPS-induced NO production and expression of iN OS in a dose-dependent manner. Additionally, MAPKs activation was also attenuated via inhibition of phosphorylation of extracellular signal-regulated kinases 1/2, c-Jun N-terminal kinase and p38 which were related to inflammatory pathway. Furthermore, HHT also effectively attenuated NF-κB activation and its translocation to the nucleus, a process that is closely linked to inflammation. LPS normally induced the expression of inflammatory cytokines such as TNF-α and IL-6, but the secretion and expression of TNF-α and IL-6 were significantly attenuated by pretreating the cells with HHT.CONCLUSION: HHT suppressed LPS-induced NO production by blocking the activation of NF-κB and MAPK signaling pathways in RAW264.7 macrophages. Furthermore, HHT may have an anti-inflammatory effect by suppressing the LPS-induced secretion of TNF-α and IL-6. Therefore, the traditional herbal formula HHT might be a useful potential therapeutic agent for inflammation.
OBJECTIVE: This study aimed to evaluate whether Hwangryunhaedoktang (HHT), a herbal compound, has an inhibitory effect on lipopolysaccharide (LPS) -induced inflammation in RAW264.7 macrophages. METHODS: The effects of HHT were evaluated by confirming nitric oxide (NO) production and expression of inducible NO synthase (iNOS) and mitogen-activated protein kinases (MAPKs) in the Griess assay, Western blotting, and real-time reverse transcription of the LPS- stimulated RAW264.7 macrophages. Western blot analyzes and luciferase assays were used to evaluate whether HHT has an effect on the phosphorylation and translocation of nuclear factor-κB (NF-κB). The secretion and expression of tumor necrosis factor-α (TNF-α) and interleukin- 6) were determined via enzyme-linked immunosorbent assay and Western blot analyzes. RESULTS: HHT suppressed LPS-induced NO production and expression of iN OS in a dose- dependent manner. Additionally, MAPKs activation was also att enuated via inhibition of phosphorylation of extracellular signal-regulated kinases 1/2, c-Jun N-terminal kinase and p38 which were related to inflammatory pathways. Furthermore, HHT also effectively attenuated NF-κB activation and its translocation to the nucleus, a process LPS normally induced the expression of inflammatory cytokines such as TNF-α and IL-6, but the secretion and expression of TNF-α and IL-6 were attenuated by pretreating the cells with HHT.CONCLUSION: HHT suppressed LPS-induced NO production by blocking the activation of NF-κB and MAPK signaling pathways in RAW264.7 macrophages. Furthermore, HHT may have an anti-inflammatory effect by suppressing the LPS-induced secretion of TNF-α and IL-6 Thus, the traditional herbal formula HHT might be a useful potential therapeutic agent for inflammation.