Medicine, Chinese traditional,Lipopolysaccharide,Nitric oxide,Nuclear factor-κB;,Mitogen-activated protein kinases,Tumor necrosis factor-α,Inflammation," /> Medicine, Chinese traditional,Lipopolysaccharide,Nitric oxide,Nuclear factor-κB;,Mitogen-activated protein kinases,Tumor necrosis factor-α,Inflammation,"/> Hwangryunhaedoktang exerts anti-inflammation on LPS-induced NO production by suppressing MAPK and NF-κB activation in RAW264.7 macrophages
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Journal of Integrative Medicine ›› 2017, Vol. 15 ›› Issue (4): 326-336.doi: 10.1016/S2095-4964(17)60350-9

• Research Article • Previous Articles    

Hwangryunhaedoktang exerts anti-inflammation on LPS-induced NO production by suppressing MAPK and NF-κB activation in RAW264.7 macrophages

Byung Hyuk Hana,b, Yun Jung Leea,b, Jung Joo Yoona,b, Eun Sik Choia,b, Seung Namgunga,b, Xian Jun Jina,b, Da Hye Jeonga,b, Dae Gill Kanga,b, Ho Sub Leea,b   

  1. a College of Oriental Medicine and Professional Graduate School of Oriental Medicine, Wonkwang University, Iksan, Jeonbuk 54538, Korea 
    b Hanbang Body-fluid Research Center, Wonkwang University, Iksan, Jeonbuk 54538, Korea
  • Received:2016-12-08 Accepted:2017-02-24 Online:2017-07-15 Published:2017-07-15
  • Contact: Dae Gill Kang, Professor; E-mail: dgkang@wku.ac.kr. Ho Sub Lee, Professor; E-mail: host@wku.ac.kr

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 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 iNOS 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.

Key words: Medicine, Chinese traditional, Lipopolysaccharide, Nitric oxide, Nuclear factor-κB;, Mitogen-activated protein kinases, Tumor necrosis factor-α, Inflammation

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