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http://dx.doi.org/10.6116/kjh.2017.32.2.77.

The study of anti-inflammatory effect of Hyeonto-dan extract in RAW 264.7 macrophage  

Kim, Ma-Ryong (Department of Oriental Pharmacy, College of Pharmacy, Wonkwang Oriental Medicines Research Institute, Wonkwang University)
Kang, Ok-Hua (Department of Oriental Pharmacy, College of Pharmacy, Wonkwang Oriental Medicines Research Institute, Wonkwang University)
Kong, Ryong (Department of Oriental Pharmacy, College of Pharmacy, Wonkwang Oriental Medicines Research Institute, Wonkwang University)
Seo, Yun-Soo (Department of Oriental Pharmacy, College of Pharmacy, Wonkwang Oriental Medicines Research Institute, Wonkwang University)
Zhou, Tian (Department of Oriental Pharmacy, College of Pharmacy, Wonkwang Oriental Medicines Research Institute, Wonkwang University)
Kim, Sang-A (Department of Oriental Pharmacy, College of Pharmacy, Wonkwang Oriental Medicines Research Institute, Wonkwang University)
Kim, Eun-Su (Department of Oriental Pharmacy, College of Pharmacy, Wonkwang Oriental Medicines Research Institute, Wonkwang University)
Sin, Min-A (Department of Oriental Pharmacy, College of Pharmacy, Wonkwang Oriental Medicines Research Institute, Wonkwang University)
Lee, Young-Seob (Department of Herbal Crop Research, National Institute of Horticultural & Herbal Science, RDA)
Kwon, Dong-Yeul (Department of Oriental Pharmacy, College of Pharmacy, Wonkwang Oriental Medicines Research Institute, Wonkwang University)
Publication Information
The Korea Journal of Herbology / v.32, no.2, 2017 , pp. 77-85 More about this Journal
Abstract
Objectives : This study aimed to investigate the unknown mechanisms behind the anti- inflammatory activity of Hyeonto-dan(HT) 70% ethanol extract on LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with Hyeonto-dan 1 h prior to addition of 200 ng/mL of LPS. Cell viability was measured by the MTS assay. Nitric oxide levels were determined by the Griess assay. $PGE_2$ were measured using EIA kit. Pro-inflammatory cytokine production was measured by the enzyme-linked immunosorbent assay (ELISA). The expression of COX-2, iNOS, and MAPKs was investigated by Western blot, qRT-PCR. $NF-{\kappa}B$/p65 localization and interaction of the TLR-4 receptor with LPS was examined by immunofluorescence assays. Results : Hyeonto-dan had no cytotoxicity at the measured concentration. Hyeonto-dan inhibited NO production and pro-inflammatory cytokines such as IL-6, $TNF-{\alpha}$, and PGE2 as well as the protein and mRNA expression of iNOS and COX-2. Moreover, Hyeonto-dan inhibited the interaction between LPS and TLR-4 in murine macrophages. It suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2), c-jun N-terminal kinase (JNK 1/2) and p38. Finally, it inhibited translocation of $NF-{\kappa}B$ in response to competitive LPS. Conclusions : Based on the results of this study, Hyeonto-dan inhibited the binding of TLR-4 receptor to LPS and inhibited the phosphorylation of extracellular signaling pathway MAPKs. These inhibitory effects are thought that the amount of $NF-{\kappa}B$ delivered to the nucleus was decreased and the inflammatory reaction was prevented by decreasing the production of LPS-induced $PGE_2$, NO, IL-6 and $TNF-{\alpha}$.
Keywords
Hyeonto-dan; Anti-inflammatory effect; RAW 264.7 cells; TLR-4;
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