[논문]당귀(當歸) 추출물이 피부 각질형성세포의 염증반응에 미치는 영향

허정; 박호연; 김엄지; 김은영; 손영주; 정혁상

doi:10.6116/kjh.2022.37.3.9.

당귀(當歸) 추출물이 피부 각질형성세포의 염증반응에 미치는 영향
Effect of Angelicae Gigantis Radix for Inflammatory Response in HaCaT Cells 원문보기

大韓本草學會誌 = The Korea journal of herbology, v.37 no.3, 2022년, pp.9 - 19

허정 (경희대학교 한의과대학 해부학교실) , 박호연 (경희대학교 한의과대학 해부학교실) , 김엄지 (경희대학교 한의과대학 해부학교실) , 김은영 (경희대학교 한의과대학 해부학교실) , 손영주 (경희대학교 한의과대학 해부학교실) , 정혁상 (경희대학교 한의과대학 해부학교실)

Abstract ▼ AI-Helper

Objectives : Angelicae Gigantis Radix (AG) is a plant of the Ranunculus family. AG have been reported to have various pharmacological effects on human health which include uterine growth promotion, anti-inflammatory, analgesic, and immune enhancement. However, research on dermatitis disease is insufficient. Therefore, we investigated the effects of AG on tumor necrosis factor-α (TNF-α)/interferon-γ (IFN-γ) stimulated HaCaT cell. Methods : To investigate the effect of AG on HaCaT cell, HaCaT cells were pre-treated with AG for 1 hour and then stimulated with TNF-α/IFN-γ. After 24 hours, media and cells were harvested to analyze the inflammatory mediators. Concentration of human interleukin-1beta (IL-1β), monocyte chemoattractant protein-1 (MCP-1), granulocyte-macrophage colony-stimulating factor (GM-CSF), and TNF-α in the media were assessed by ELISA. mRNA expression of human thymus and activation-regulated chemokine (TARC), IL-6, and IL-8 were analyzed by RT-PCR. Additionally, the mechanisms of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway were investigated by Western blot. Results : The treatment of AG inhibited gene expression levels of IL-6, IL-8, and TARC and protein expression levels of IL-1β, MCP-1, and GM-CSF. Also, AG significantly reduced extracellular signal-regulated kinase (ERK) phosphorylation and NF-κB translocation in TNF-α/IFN-γ stimulated HaCaT cell. Conclusions : Taken together, these results demonstrate that AG can alleviate inflammatory diseases such as atopic dermatitis by regulating the expression of inflammatory cytokines. Also, it suggest that AG may a promising candidate drug for the treatment of inflammatory disease such as atopic dermatitis.

주제어

표/그림 (7)

표 Table 1. Primer Sequence for RT-PCR Analysis.
그림 Figure 1. Cell viability of AG in HaCaT cells. Cell viability was determined with MTS assay. Cells were treated with AG from 15.6 to 500 μg/ml for 24 h. Values are mean ± SEM of three independent experiments. ^#p < 0.05 and ^##p < 0.01 vs. Nor.
표 Table 2. Summary of Antibodies Used in Western Blot
그림 Figure 2. Effects of AG on TARC, IL-6, and IL-8 mRNA expression levels in HaCaT cells. Cells were pre-treated with various concentration of AG for 1 h prior to treated with TNF-α/IFN-γ (10 ng/ml) for 24 h. (A) mRNA expression of IL-6, IL-8, and TARC were examined by RT-PCR. (B-D) mRNA expressions were measured using Image J software. Values are mean ± SEM of three independent experiments. ^##p < 0.01 vs. TNF-α/IFN-γ (－) and *p < 0.05, **p < 0.01 vs. TNF-α/IFN-γ (＋).
그림 Figure 3. Effects of AG on IL-1β, MCP-1, GM-CSF, and TNF-α protein expression levels in HaCaT cells. Cells were pre-treated with various concentration of AG for 1 h prior to treated with TNF-α/IFN-γ (10 ng/ml) for 24 hours. (A-D) Protein expressions of IL-1β, MCP-1, GM-CSF, and TNF-α were measured by ELISA. Values are mean ± SEM of three independent experiments. ^##p < 0.01 vs. TNF-α/IFN-γ (－) and *p < 0.05, **p < 0.01 vs. TNF-α/IFN-γ (＋).
그림 Figure 4. Effects of AG on phosphorylation of ERK, JNK, and p38 in HaCaT cells. Cells were pre-treated with various concentration of AG for 1 h prior to treated with TNF-α/IFN-γ (10 ng/ml) for indicated times. (A) Phosphorylation of MAPKs were detected by Western blot. (B-D) Phosphorylated-protein expressions were measured using Image J software. Values are mean ± SEM of three independent experiments. ^#p < 0.05, ^##p < 0.01 vs. TNF-α/IFN-γ (－) and *p < 0.05 vs. TNF-α/IFN-γ (＋).
그림 Figure 5. Effects of AG on translocation of NF-κB in HaCaT cells. Cells were pre-treated with various concentration of AG for 1 h prior to treated with TNF-α/IFN-γ (10 ng/ml) for indicated times. (A) The degree of NF-κB protein in nucleus and IκBα protein in cytosol were analyzed by Western blot. (B-D) NF-κB and IκBα expressions were measured using Image J software. Values are mean ± SEM of three independent experiments. ^#p < 0.05, ^##p < 0.01 vs. TNF-α/IFN-γ (－) and *p < 0.05, **p < 0.01 vs. TNF-α/IFN-γ (＋).

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