[논문]한국산 겨우살이의 항염증 효과

홍창의; 임완택; 유수연

doi:10.15230/scsk.2022.48.3.265

한국산 겨우살이의 항염증 효과
Anti-inflammatory Activity of Viscum album var. coloratum In Vitro 원문보기

大韓化粧品學會誌 = Journal of the society of cosmetic scientists of Korea, v.48 no.3, 2022년, pp.265 - 273

홍창의 (순천대학교 약학과) , 임완택 (순천대학교 약학과) , 유수연 (순천대학교 약학과 및 생명약학연구소)

초록
AI-Helper

본 연구에서는 한국산 겨우살이 (Viscum album L. var. coloratum)이 아토피 피부염과 관련된 염증성 사이토카인에 영향을 미치는지 여부를 알아보았다. 실험에는 헥산, 부탄올, 에틸아세테이트, 메틸렌클로라이드, 총 4 가지 분획물을 사용하였으며, RAW264.7 마우스 대식세포와 RBL-2H3 렛트 호중구를 이용하여 염증성 마커를 연구하였다. 실험 결과 에틸아세테이트 분획이 tumor necrosis factor alpha (TNF-α), interleukin(IL)-6, IL-4의 mRNA 발현 및 단백질 분비량을 감소시켰으나, 헥산 분획은 뚜렷한 효능이 없었다. 또한 부탄올 분획은 IL-4, IL-6의 mRNA 발현을 감소시켰고, 메틸렌클로라이드 분획은 IL-4와 TNF-α의 mRNA 발현을 감소시켰다. 결과적으로 한국산 겨우살이(V. album var. coloratum)가 아토피 피부염과 관련된 사이토카인 분비를 억제시켜 항염증 효과를 나타낼 수 있으므로, 이와 관련된 기능성 화장품 개발이 가능할 것으로 사료된다.

Abstract ▼ AI-Helper

In this study, we investigated whether Korean mistletoe (Viscum album L. var. coloratum) has anti-inflammatory effects that play key roles in the regulation of pathological mechanism of atopic dermatitis (AD). Four kinds of fractions, hexane (HX), ethyl acetate (EA), butanol (BU), and methylene chloride (MC), were used and RAW264.7 mouse macrophages and RBL-2H3 rat basophils were used to measure various inflammatory markers. EA significantly decreased mRNA expression and protein secretion levels of tumor necrosis factor alpha (TNF-α), interleukin (IL)-6, and IL-4 but HX did not affect these markers. In addition, BU decreased mRNA expressions of IL-4 and IL-6 whereas MC decreased IL-6 and TNF-α mRNA expressions. As a result, Korean mistletoe can show anti-inflammatory effects by inhibiting the secretion of cytokines related to AD, so it is thought that it will be possible to develop functional cosmetics related to this.

주제어

표/그림 (7)

$Figure 1. Effect of different fractions of Korean mistletoe (Viscum album var. coloratum) on cell viability. RAW 264.7 cells and RBL-2H3 cells were treated with different concentration of V. album coloratum fractions for 48 h. Cell viability was measured by MTT assay. Viability of untreated control cells was defined as 100% and the values are expressed as the means ± SD (N = 3). (HX; hexane fraction, EA; ethylene acetate fraction, BU; butanol fraction, MC; methylene chloride fraction).$ 그림 Figure 1. Effect of different fractions of Korean mistletoe (Viscum album var. coloratum) on cell viability. RAW 264.7 cells and RBL-2H3 cells were treated with different concentration of V. album coloratum fractions for 48 h. Cell viability was measured by MTT assay. Viability of untreated control cells was defined as 100% and the values are expressed as the means ± SD (N = 3). (HX; hexane fraction, EA; ethylene acetate fraction, BU; butanol fraction, MC; methylene chloride fraction).
표 Table 1. Primers for Quantitative Real-time PCR
$Figure 2. Effect of different fractions of V. album coloratum on the production of NO in LPS-stimulated RAW 264.7 cells. Cells were co-treated with different fractions of V. album coloratum (100 μg/mL) and LPS (100 ng/mL) for 24 h. The treated culture media was collected and assayed for NO production using Griess reagent. Each bar represent mean ± SD (N = 3). ( *** represents significant differences between LPS and samples treated group p < 0.001, **p < 0.01, *p < 0.05). (HX; hexane fraction, EA; ethylene acetate fraction, BU; butanol fraction, MC; methylene chloride fraction).$ 그림 Figure 2. Effect of different fractions of V. album coloratum on the production of NO in LPS-stimulated RAW 264.7 cells. Cells were co-treated with different fractions of V. album coloratum (100 μg/mL) and LPS (100 ng/mL) for 24 h. The treated culture media was collected and assayed for NO production using Griess reagent. Each bar represent mean ± SD (N = 3). ( *** represents significant differences between LPS and samples treated group p < 0.001, **p < 0.01, *p < 0.05). (HX; hexane fraction, EA; ethylene acetate fraction, BU; butanol fraction, MC; methylene chloride fraction).
$Figure 3. Effect of different fractions of V. album coloratum on the production of PGE<sub>2</sub> in LPS-stimulated RAW 264.7 cells. Cells were pretreated with different fractions of V. album coloratum (100 μg/mL) for 1 h, followed by stimulation with LPS (100 ng/mL) for 24 h. The treated culture media was collected and assayed for PGE2 production using ELISA kit. Each bar represent mean ± SD (N = 3). ( *** represents significant differences between LPS and samples treated group p < 0.001, **p < 0.01, *p < 0.05). (HX; hexane fraction, EA; ethylene acetate fraction, BU; butanol fraction, MC; methylene chloride fraction).$ 그림 Figure 3. Effect of different fractions of V. album coloratum on the production of PGE₂ in LPS-stimulated RAW 264.7 cells. Cells were pretreated with different fractions of V. album coloratum (100 μg/mL) for 1 h, followed by stimulation with LPS (100 ng/mL) for 24 h. The treated culture media was collected and assayed for PGE2 production using ELISA kit. Each bar represent mean ± SD (N = 3). ( *** represents significant differences between LPS and samples treated group p < 0.001, **p < 0.01, *p < 0.05). (HX; hexane fraction, EA; ethylene acetate fraction, BU; butanol fraction, MC; methylene chloride fraction).
$Figure 4. Effect of different fractions of V. album coloratum on the production of (A) TNF-α, (B) IL-6 in LPS-stimulated RAW 264.7 cells, and (C) IL-4 in DNP-stimulated RBL-2H3 cells. RAW 264.7 cells were co-incubation with different fractions of V. album coloratum (100 μg/mL) and LPS (100 ng/mL) for 24 h. RBL-2H3 cells were incubation with DNP-IgE (500 ng/mL) for 24 h, followed by co-incubation with DNP-BSA (100 ng/mL) and different fractions of Korean mistletoe (100 μg/mL) for 4 h. The treated culture media was collected and assayed for TNF-α, IL-6, and IL-4 production using ELISA kit. Each bar represent mean ± SD (N = 3). ( *** represents significant differences between LPS and samples treated group p < 0.001, **p < 0.01, *p < 0.05). (HX; hexane fraction, EA; ethylene acetate fraction, BU; butanol fraction, MC; methylene chloride fraction).$ 그림 Figure 4. Effect of different fractions of V. album coloratum on the production of (A) TNF-α, (B) IL-6 in LPS-stimulated RAW 264.7 cells, and (C) IL-4 in DNP-stimulated RBL-2H3 cells. RAW 264.7 cells were co-incubation with different fractions of V. album coloratum (100 μg/mL) and LPS (100 ng/mL) for 24 h. RBL-2H3 cells were incubation with DNP-IgE (500 ng/mL) for 24 h, followed by co-incubation with DNP-BSA (100 ng/mL) and different fractions of Korean mistletoe (100 μg/mL) for 4 h. The treated culture media was collected and assayed for TNF-α, IL-6, and IL-4 production using ELISA kit. Each bar represent mean ± SD (N = 3). ( *** represents significant differences between LPS and samples treated group p < 0.001, **p < 0.01, *p < 0.05). (HX; hexane fraction, EA; ethylene acetate fraction, BU; butanol fraction, MC; methylene chloride fraction).
$Figure 5. Realtime PCR analysis of mRNA expression of (A) TNF-α, (B) IL-6 in LPS-stimulated RAW 264.7 cells, and (C) IL-4 in DNP-stimulated RBL-2H3 cells. RAW 264.7 cells were co-incubated with different concentration of V. album coloratum fractions and LPS (100 ng/mL). RBL-2H3 cells were incubated with DNP-IgE (500 ng/mL) for 24 h, followed by co-incubation with DNP-BSA (100 ng/mL) and different concentration of V. album coloratum fractions for 4 h. Each bar represent mean ± SD (N = 3). ( *** represents significant differences between LPS and samples treated group p < 0.001, **p < 0.01, *p < 0.05). (HX; hexane fraction, EA; ethylene acetate fraction, BU; butanol fraction, MC; methylene chloride fraction).$ 그림 Figure 5. Realtime PCR analysis of mRNA expression of (A) TNF-α, (B) IL-6 in LPS-stimulated RAW 264.7 cells, and (C) IL-4 in DNP-stimulated RBL-2H3 cells. RAW 264.7 cells were co-incubated with different concentration of V. album coloratum fractions and LPS (100 ng/mL). RBL-2H3 cells were incubated with DNP-IgE (500 ng/mL) for 24 h, followed by co-incubation with DNP-BSA (100 ng/mL) and different concentration of V. album coloratum fractions for 4 h. Each bar represent mean ± SD (N = 3). ( *** represents significant differences between LPS and samples treated group p < 0.001, **p < 0.01, *p < 0.05). (HX; hexane fraction, EA; ethylene acetate fraction, BU; butanol fraction, MC; methylene chloride fraction).
$Figure 6. Effect of different fractions of V. album coloratum on the expression of iNOS and COX-2 in LPS-stimulated RAW 264.7 cells. Cells were pretreated with different concentration of V. album coloratum fractions (1 h), followed by stimulation with LPS (100 ng/mL, 18 h) to determine the expression of iNOS and COX-2. Each bar represent mean ± SD (N = 3). ( *** represents significant differences between LPS and samples treated group p < 0.001, **p < 0.01, *p < 0.05). (HX; hexane fraction, EA; ethylene acetate fraction, BU; butanol fraction, MC; methylene chloride fraction).$ 그림 Figure 6. Effect of different fractions of V. album coloratum on the expression of iNOS and COX-2 in LPS-stimulated RAW 264.7 cells. Cells were pretreated with different concentration of V. album coloratum fractions (1 h), followed by stimulation with LPS (100 ng/mL, 18 h) to determine the expression of iNOS and COX-2. Each bar represent mean ± SD (N = 3). ( *** represents significant differences between LPS and samples treated group p < 0.001, **p < 0.01, *p < 0.05). (HX; hexane fraction, EA; ethylene acetate fraction, BU; butanol fraction, MC; methylene chloride fraction).

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표제어: PCR

동의어: Packet Collision Rate

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용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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