玄兎丹의 RAW 264.7 대식 세포에서의 항염증 효과에 관한 연구

• Published in: 대한본초학회지, 32(2) pp. 77 - 85
• Main Authors: 김마룡, 강옥화, 공룡, 서윤수, 주전, 김상아, 김은수, 신민아, 이영섭, 권동렬
• Format: Journal Article
• Language: Korean
• Published: 대한본초학회 01.03.2017
• Subjects: 한의학
• ISSN: 1229-1765; 2288-7199
• DOI: 10.6116/kjh.2017.32.2.77

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. PGE2 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-κB/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-α, 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-κB 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-κB delivered to the nucleus was decreased and the inflammatory reaction was prevented by decreasing the production of LPS-induced PGE2,NO, IL-6 and TNF-α. KCI Citation Count: 0