[논문]금은화산(金銀花散)의 LPS로 유도된 Raw 264.7 세포에서의 항염증 효과

김연수; 김수진; 지선영; 황보민

doi:10.6114/jkood.2021.34.3.001

금은화산(金銀花散)의 LPS로 유도된 Raw 264.7 세포에서의 항염증 효과
Anti-inflammatory Effect of Geumeunwha-san Water Extract on LPS-induced Raw 264.7 Cells 원문보기

韓方眼耳鼻咽喉皮膚科學會誌 = The journal of Korean Medicine Ophthalmology & Otolaryngology & Dermatology, v.34 no.3, 2021년, pp.1 - 12

김연수 (대구한의대 부속 대구한방병원 안이비인후피부과) , 김수진 (대구한의대 부속 대구한방병원 안이비인후피부과) , 지선영 (대구한의대 부속 포항한방병원 안이비인후피부과) , 황보민 (대구한의대 부속 대구한방병원 안이비인후피부과)

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Objectives : The purpose of this study is to investigate the anti-inflammatory effect of Geumeunwha-san(GEHS) water extract in vitro. Methods : To evaluate the anti-inflammatory effect of GEHS, Raw 264.7 cells were pretreated with 10-300㎍/㎖ of GEHS for 1hr, and then exposed to 1㎍/㎖ of LPS. MTT assay was used to detect the cell viability. Productions of pro-inflammatory cytokines and NO were measured in culture media. By using immunoblot analysis, protein levels of iNOS and NF-𝜅B were determined. Results : In vitro study, cell viability assay on GEHS treatment of 10-300㎍/㎖ has no cytotoxicity in Raw 264.7 cells. Pretreated 100, 300㎍/㎖ of GEHS had significantly inhibited LPS-induced NO production. And also pretreatment of 100, 300㎍/㎖ GEHS had significantly decreased production of interleukin-6, -1𝛽 and tumor necrosis factor-𝛼 in LPS-activated Raw 264.7 cells. In addition, GEHS reduced LPS-mediated iNOS expression. Moreover I-𝜅B𝛼 expression was significantly induced by GEHS and NF-𝜅B expression was reduced by GEHS. Conclusions : These results suggest the clinical basis of GEHS for the treatment of inflammatory diseases.

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표/그림 (6)

표 Table 3. The Composition of Geumeunwha-san
그림 Fig. 1. Effect of GEHS on NO Production in LPS-activated Raw 264.7 Cells. 10-300㎍/㎖ of GEHS were treated on Raw 264.7 cells, dissolved in DMEM for 1hr before the treatment of LPS(1㎍/㎖), and then the cells were incubated for 18hr. The concentration of nitric oxide in culture medium was evaluated as described in materials and methods part. Values represent the mean±SD with separate experiments. ** p<0.01. ## p<0.01.
그림 Fig. 2. Effect of GEHS on Cell Viability. Raw 264.7 cells were treated with 10, 30, 100, 300㎍/㎖ of GEHS dissolved in DMEM for 1hr before the treatment of LPS(1㎍/㎖), and then the cells were incubated for 18hr. After the incubation, the cells were treated with MTT solution. Values represent the mean ± SD with each independent experiments. ** p<0.01.
그림 Fig. 3. Effect of GEHS on iNOS Expression in LPS-activated Raw 264.7 Cells. iNOS protein’s levels were evaluated 18hr after the treatment of LPS(1㎍/㎖) with or without GEHS(100, 300㎍/㎖) pretreatment(i.e. 1hr before LPS). SDS-PAGE separates equal amounts of protein. β-actin was used as an equal loading control (A). The protein’s relative density level was measured by scanning densitometry (B). Values represent the mean ± SD for each condition. ** p<0.01, ## p<0.01.
그림 Fig. 4. Effect of GEHS on Cytokine Production in LPS-activated Raw 264.7 Cells. 100, 300㎍/㎖ of GEHS were treated on Raw 264.7 cells for 1hr, and then incubated with 1㎍/㎖ of LPS for 18hr. (A) Production of TNF-α, (B) Production of IL-1β, (C) Production of IL-6 were examined by ELISA. Values represent mean ± SD of each experiments. Significant vs untreated controls, **p<0.01; significant vs LPS-stimulated cells, ##p<0.01.
그림 Fig. 5. Effect of GEHS on NF-κB Signaling Pathway. 100, 300㎍/㎖ of GEHS were treated on Raw 264.7 cells for 30(IκBα) or 60(p-IκBα, NF-κB)min, and then incubated with 1㎍/㎖ of LPS(i.e. 1hr before LPS). (A, B)Western blot was used to examine the relative levels of I-κBα, (A, C)p-IκBα protein and (D, E)NF-κB protein. Values represent mean ± SD of each experiments. Significant vs untreated control, **p<0.01; significant vs LPS-stimulated cells, ##p<0.01.

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금은화산(金銀花散)의 LPS로 유도된 Raw 264.7 세포에서의 항염증 효과
Anti-inflammatory Effect of Geumeunwha-san Water Extract on LPS-induced Raw 264.7 Cells 원문보기

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금은화산(金銀花散)의 LPS로 유도된 Raw 264.7 세포에서의 항염증 효과 Anti-inflammatory Effect of Geumeunwha-san Water Extract on LPS-induced Raw 264.7 Cells 원문보기

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금은화산(金銀花散)의 LPS로 유도된 Raw 264.7 세포에서의 항염증 효과
Anti-inflammatory Effect of Geumeunwha-san Water Extract on LPS-induced Raw 264.7 Cells 원문보기

표/그림 (6)

표/그림 (6)