[논문]침엽수 4종 목부 정유의 항염증 효과 평가

YANG, Jiyoon; CHOI, Won-Sil; KIM, Jae-Woo; LEE, Sung-Suk; PARK, Mi-Jin

doi:10.5658/wood.2019.47.6.674

침엽수 4종 목부 정유의 항염증 효과 평가
Anti-Inflammatory Effect of Essential Oils Extracted from Wood of Four Coniferous Tree Species 원문보기

목재공학 = Journal of the Korean wood science and technology, v.47 no.6, 2019년, pp.674 - 691

YANG, Jiyoon (Division of Wood Chemistry, Department of Forest Products, National Institute of Forest Science) , CHOI, Won-Sil (Division of Life Sciences, School of Life Sciences, Korea University) , KIM, Jae-Woo (Division of Wood Chemistry, Department of Forest Products, National Institute of Forest Science) , LEE, Sung-Suk (Division of Wood Chemistry, Department of Forest Products, National Institute of Forest Science) , PARK, Mi-Jin (Division of Wood Chemistry, Department of Forest Products, National Institute of Forest Science)

초록
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본 연구는 국내산 편백, 소나무, 잣나무, 일본잎갈나무 목부 정유의 항염증 효과를 평가 하였다. 실험에 사용된 정유는 각 수종의 목부에서 증류법(hydrodistillation)으로 추출되었다. 추출한 정유들을 GC-MS를 통해 성분 분석한 결과, 편백의 경우 ${\alpha}$-cadinol (19.25%), ${\tau}$-muurolol (14.20%) 및 ${\alpha}$-pinene (13.74%)이, 소나무는 ${\alpha}$-pinene (47.16%), longifolene (14.31%), ${\beta}$-phellandrene (11.78%) 및 ${\beta}$-pinene (11.02%)이 주요성분이었으며, 잣나무의 경우 ${\alpha}$-pinene (13.49%)와 longifolene(10.79%), 낙엽송의 경우 geranyl linalool (23.58%) 및 ${\alpha}$-pinene (18.57%)이 주요성분으로 확인되었다. 침엽수 목부 정유들의 항염증 효과는 LPS(Lipopolysaccharide)로 염증반응이 유도된 RBL-2H3 비만세포주에 정유를 처리한 후 IL-4(interleukin-4), IL-13(interleukin-13), ${\beta}$-hexosaminidase의 발현 변화를 조사함으로써 평가하였다. MTT assay를 통한 세포 독성 평가 결과, 침엽수 4종 정유 모두 $10^{-7}{\sim}10^{-5}%$ 농도에서 세포 독성을 보이지 않았다. 염증반응이 유도된 RBL-2H3 세포에 침엽수 4종의 목부 정유를 $10^{-7}%$ 농도로 처리하였을 때, LPS 처리군 대비 침엽수 4종의 목부 처리군에서 IL-4와 IL-13의 상대적 발현량이 큰 폭으로 감소하였으며, ${\beta}$-hexosaminidase 분비가 유의하게 억제되었다. 침엽수 정유 4종의 IL-4 상대적 발현량은 LPS 처리군 대비 소나무 69.6%, 잣나무 63.2%, 편백 55.1% 그리고 낙엽송 45.8% 감소하였다. IL-13의 상대적 발현량은 소나무 57.8%, 잣나무 57.1%, 편백 51.1% 그리고 낙엽송 34.5% 감소하였다. ${\beta}$-Hexosaminidase 분비의 경우 LPS 처리군 대비 편백 38.1%, 소나무 33.0%, 잣나무 27.4% 그리고 낙엽송 9.1% 억제하였다. Il-4, IL-13 및 ${\beta}$-hexosaminidase 분비를 비교한 결과, 침엽수 목부 정유 4종 중 소나무 정유의 활성이 가장 우수하였다. 이러한 결과는 국내 침엽수 4종의 목부 정유는 염증반응을 유발하는 싸이토카인의 발현 억제를 통하여 항염증 효과를 나타냄을 확인할 수 있었다.

Abstract ▼ AI-Helper

The aim of this study was to evaluate the anti-inflammatory effects of essential oils extracted from the wood of Chamaecyparis obtusa, Pinus densiflora, Pinus koraiensis, and Larix kaempferi. Essential oils were extracted by hydrodistillation, and their chemical components were determined by GC/MS. Major chemical components of these essential oils were ${\alpha}$-cadinol (19.25%), ${\tau}$-muurolol (14.20%), and ${\alpha}$-pinene (13.74%) in C. obtusa; ${\alpha}$-pinene (47.16%), longifolene (14.31%), ${\beta}$-phellandrene (11.78%), and ${\beta}$-pinene (11.02%) in P. densiflora; ${\alpha}$-pinene (13.49%) and longifolene (10.79%) in P. koraiensis, and geranyl linalool (23.58%) and ${\alpha}$-pinene (18.57%) in L. kaempferi. To evaluate the anti-inflammatory effects of essential oils, lipopolysaccharide (LPS)-induced RBL-2H3 mast cells were treated with these essential oils; then, the changes in the mRNA expression level of the cytokines IL-4 and IL-13 were examined. Further, degranulation was evaluated by measuring ${\beta}$-hexosaminidase release. After LPS-induced RBL-2H3 mast cells were exposed to $10^{-7}%$ of all types of essential oils, the gene expression levels of IL-4 and IL-13 within the cells remarkably decreased. The relative mRNA expression level of IL-4 was 69.6% in P. densiflora, 63.2% in P. koraiensis, 55.1% in C. obtusa, and 45.8% in L. kaempferi compared with that in the group treated with LPS. The mRNA expression level of L-13 should a similar trend. The inhibitory rate of IL-13 mRNA expression of P. densiflora, P. koraiensis, C. obtusa, and L. kaempferi was 57.8%, 57.1%, 51.1%, and 34.5%, respectively. ${\beta}$-Hexosaminidase release significantly decreased following the treatment with the four types of essential oils. The rate of ${\beta}$-hexosaminidase release were 38.1% C. obtusa; 33.0% P. densiflora; 27.4% P. koraiensis; and 9.1% L. kaempferi. Among all types of essential oils, that extracted from P. densiflora wood showed the highest anti-inflammatory activity. These results show that the tested essential oils exert an anti-inflammatory effect through the inhibition of degranulation and expression of cytokines.

주제어

표/그림 (5)

표 Table 1. Oligonucleotide sequences for quantitative real-time polymerase chain reaction
표 Table 2. The major constituents of four coniferous oils
그림 Fig. 1. Effects of four coniferous oils on the viability of RBL-2H3 cells. Group: VE(vehicle), treatment of 10-7~10-5% essential oil(C. obtusa, P. densiflora, P. koraiensis, L. kaempferi). Values are expressed as the mean±standard deviation.
그림 Fig. 2. Pro-inflammatory cytokine mRNA expression on RBL-2H3 cells treated with four coniferous oils. (a) mRNA expression level of IL-4, (b) mRNA expression level of IL-13. VE : vehicle, NC : negative control(group treated with lipopolysaccharide), PC : positive control(group treated with dexamethasone on lipopolysaccharideinduced RBL-2H3 cells), treatment of essential oil(C. obtusa, P. densiflora, P. koraiensis, L. kaempferi). Values are expressed as the mean±standard deviation. *P<0.05 vs. VE(vehicle); #P<0.05 vs. NC (lipopolysaccharidetreated group).
그림 Fig. 3. Inhibitory effects of four coniferous oils on the release of β-hexosaminidase. VE : vehicle, NC : negative control(group treated with lipopolysaccharide), PC : positive control(group treated with dexamethasone on lipopolysaccharide-induced RBL-2H3 cells), treatment of essential oil(C. obtusa, P. densiflora, P. koraiensis, L. kaempferi). Values are expressed as the mean±standard deviation. *P<0.05 vs. VE(vehicle); #P<0.05 vs. NC(lipopolysaccharide-treated group).

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문제 정의

kaempferi), all of which are typical local tree species used for timber. This study also aimed to verify the potential use of essential oils extracted from coniferous wood for improving or preventing allergic diseases.

제안 방법

There are varied ongoing studies on the bioactivity of essential oils extracted from coniferous tree species, but limited to leaves and the studies on the bioactivity of essential oils extracted from the wood are insufficient. Therefore, this study aimed to compare and analyze the anti-inflammatory activity of essential oils extracted from the wood of four coniferous tree species including cypress (C. obtusa), pine (P. densiflora), pine nut (P. koraiensis), and Japanese larch (L. kaempferi), all of which are typical local tree species used for timber. This study also aimed to verify the potential use of essential oils extracted from coniferous wood for improving or preventing allergic diseases.
To determine the influence of essential oils extracted from the wood of the four coniferous tree species on the survival and reproduction of RBL-2H3 mast cells, these essential oils were processed at concentrations of 10−7%–10−5%, and then MTT assay was performed.

대상 데이터

The essential oils used in the experiments were extracted from grinded wood of the four coniferous tree species using hydrodistillation. Determination of their chemical composition using GC/MS revealed the prominent substances to be α-pinene, β-pinene, fenchol, and α-terpineol in all coniferous tree species, wherein each tree species showed different compositions of these major substances
To assess the level of anti-inflammatory effect of essential oils extracted from the wood of the four coniferous tree species, RBL-2H3 mast cells (CRL22256™; American Type Culture Collection, Manassas, Manassas, VA, USA) were purchased in lots.

성능/효과

Assessment of IL-4 and IL-13 gene expressions and β-hexosaminidase release rate in terms of inflammation, the relative IL-4 and IL-13 gene expressions stimulated by LPS decreased substantially following the application of essential oils extracted from the wood of pine and pine nut, whereas the highest inhibition of β-hexosaminidase release was observed for cypress.
Examination of the effect of the essential oils that affect β-hexosaminidase release, the essential oil extracted from cypress wood showed the highest inhibition rate, and the relatively high percentage of τ-muurolol and αcadinol was remarkable.
Thus, further research on the bioactive substances responsible for the anti-inflammatory effect is warranted. Through this study, the essential oils extracted from the wood of the four coniferous tree species were found to be effective in improving allergic inflammation. The bioactive substances in essential oils are diffused in air when cypress, pine, pine nut and Japanese larch trees are used as wood materials for interior construction.

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