[논문]Raw 264.7 세포에서 화독단(化毒丹)의 항염증 효능에 대한 연구

신기석; 황보민; 지선영

doi:10.6114/jkood.2022.35.3.036

Raw 264.7 세포에서 화독단(化毒丹)의 항염증 효능에 대한 연구
Research of the Anti-inflammatory Effects of Hwadokdan on 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.35 no.3, 2022년, pp.36 - 47

신기석 (대구한의대학교 한의과대학 안이비인후피부과학교실) , 황보민 (대구한의대학교 한의과대학 안이비인후피부과학교실) , 지선영 (대구한의대학교 한의과대학 안이비인후피부과학교실)

Abstract ▼ AI-Helper

Objectives : The purpose of this study was to evaluate the anti-Inflammatory effect of Hwadokdan(HDD) extracted with water. Methods : We pretreated LPS-activated Raw 264.7 cell with HDD and anti-inflammatory effect of HDD is investigated by measuring NO production, relative iNOS, and TNF-α, IL-6, IL-1β, PGE2 productions level, p-IκBα, IκBα, NFκB. Results : HDD reduced NO production and iNOS protein, TNF-α, IL-6 level at concentration of 100㎍/㎖ and 300㎍/㎖ in LPS-activated Raw 264.7 cell significantly. There wasn't a significant cytotoxicity in MTT assay. And HDD reduced IL-1β level at concentration of 300㎍/㎖ in LPS-activated Raw 264.7 cell significantly. HDD reduced p-IκBα and NFκB protein level at concentration of 300㎍/㎖ in LPS-activated Raw 264.7 cell significantly. And HDD enhanced IκBα protein level at concentration of 100㎍/㎖ and 300㎍/㎖ in LPS-activated Raw 264.7 cell significantly. Conclusions : These suggests that HDD can be used as a therapeutic drug for various inflammatory diseases.

주제어

표/그림 (6)

표 Table 1. The Composition of Hwadokdan
그림 Fig. 1. Effects of HDD on NO Production in LPS-Activated Raw 264.7 Cells. Raw 264.7 cells were pre-treated with 10-300㎍/㎖ of HDD dissolved in DMEM for 1 hour before the LPS(1㎍/ ㎖) treatment, and then the cells were cultivated for 18 hours. The concentration of nitric oxide in medium was assessed as delineated in materials and methods part. Values represent the mean±SD with four separate experiments for each condition. *: vs. control, ** p<0.01, #: vs. LPS alone, ## p<0.01
그림 Fig. 2. Effects of HDD on Cell Viability in LPS-Activated Raw 264.7 Cells. Raw 264.7 cells were pre-treated with 10-300㎍/㎖ of HDD dissolved in DMEM for 1 hour before the LPS(1㎍/ ㎖) treatment, and then the cells were cultivated for 18 hours. And MTT solution was treated to the cultivated cells. Values represent the mean±SD with four separate experiments for each condition. *: vs. control, ** p<0.01
그림 Fig. 3. Effects of HDD on iNOS Expression in LPS-Activated Raw 264.7 Cells. Raw 264.7 cells were pre-treated with 100, 300㎍/㎖ of HDD dissolved in DMEM for 1 hour before the LPS(1㎍/㎖) treatment, and then the cells were cultivated for 18 hours. The levels of iNOS protein was assessed as delineated in materials and methods part. (A) β-actin was used as an equal loading control. (B) The relative density level of the protein was measured by scanning densitometry. Values represent the mean±SD with four separate experiments for each condition. *: vs. control, ** p<0.01, #: vs. LPS alone, ## p<0.01
그림 Fig. 4. Effects of HDD on Cytokine Production in LPS-Activated Raw 264.7 Cells. Raw 264.7 cells were pre-treated with 100, 300㎍/㎖ of HDD for 1 hour before the LPS(1㎍/㎖) treatment, and then the cells were cultivated for 18 hours. The amount of (A)TNF-α, (B)IL-1β, (C)IL-6, (D)PGE₂ was measured by ELISA kit. Values represent the mean±SD with three separate experiments for each condition. *: vs. control, ** p<0.01, #: vs. LPS alone, ## p<0.01
그림 Fig. 5. Effects of HDD on IκBα, p-IκBα and NFκB Expression in LPS-Activated Raw 264.7 Cells. Raw 264.7 cells were pre-treated with 100, 300㎍/㎖ of HDD dissolved in DMEM for 1 hour before the LPS(1㎍/㎖) treatment, and then cultivated for 30(IκBα) or 60(p-IκBα, NFκB) minutes. (A, D) β-actin and Lamin A/C were used as an equal loading control. (B, C, E) The relative density level of the IκBα, p-IκBα and NFκB was measured by scanning densitometry. Values represent the mean±SD with four separate experiments for each condition. *: vs. control, ** p<0.01, #: vs. LPS alone, ## p<0.01

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