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침엽수 4종 목부 정유의 항염증 효과 평가

Anti-Inflammatory Effect of Essential Oils Extracted from Wood of Four Coniferous Tree Species

  • 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)
  • 투고 : 2019.07.11
  • 심사 : 2019.10.16
  • 발행 : 2019.11.25

초록

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.

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.

키워드

참고문헌

  1. Ahn, C., Park, M., Kim, J., Yang, J., Lee, S., Jeung, E. 2018. Cytotoxic evaluation of plant essential oils in human skin and lung cells. Journal of the Korean Wood Science and Technology 46(2): 166-177. https://doi.org/10.5658/WOOD.2018.46.2.166
  2. Carvalho-Freitas, M.I.R., Costa, M. 2002. Anxiolytic and sedative effects of extracts and essential oil from Citrus aurantium L. Biological and Pharmaceutical Bulletin 25(12): 1629-1633. https://doi.org/10.1248/bpb.25.1629
  3. Cho, E., Ju, S., Cho, H., Kim, M., Choi, I., Choi, Y. 2009. Inhibitory effects of ethanol extracts from pine buds (Pinus densiflora) on angiotensin converting enzyme, xanthine oxidase and nitric oxide synthesis. Journal of Life Science 19(11): 1629-1636. https://doi.org/10.5352/JLS.2009.19.11.1629
  4. Chon, S., Shin, M.Y. 1999. Effects of local climatic condition on the early growth in Korean white pine (Pinus koraiensis Sieb. et Zucc.) stands. Korean Journal of Agricultural and Forest Meteorology 1(1): 1-11.
  5. Han, Y., Kim, M., Lee, H., Kang, J., Eom, C. 2017. Comparison of cellular anatomical, physical and mechanical properties between Dahurian larch and Japanese larch. Journal of the Korean Wood Science and Technology 45(5): 525-534. https://doi.org/10.5658/WOOD.2017.45.5.525
  6. Ham, Y., Kim, T. 2018. Plant extracts inhibiting biofilm formation by Streptococcus mutans without antibiotic activity. Journal of the Korean Wood Science and Technology 46(6): 692-702. https://doi.org/10.5658/WOOD.2018.46.6.692
  7. Hershey, G.K., FriedrichM, F.L., Esswein, A., Thomas, M.L., Chatila, T.A. 1997. The association of atopy with a gain-of-function mutation in the a subunit of the interleukin-4 receptor. The New England Journal of Medicine 337(24): 1720-1725. https://doi.org/10.1056/NEJM199712113372403
  8. Hong, E., Na., K., Choi, I., Choi, K., Jeung, E. 2004. Antibacterial and antifungal effects of essential oils from coniferous trees. Biological and Pharmaceutical Bulletin 27(6): 863-866. https://doi.org/10.1248/bpb.27.863
  9. Howard, T.D., Whittaker, P.A., Zaiman, A.I., Koppelman, G.H., Xu, J., Hanley, M.T., Meyer, D.A., Postma, D.S., Bleecker, E.R. 2001. Identification and association of polymorphisms in the interleukin-13 gene with asthma and atopy in a Dutch population. American Journal of Respiratory Cell and Molecular Biology 25(3): 377-384. https://doi.org/10.1165/ajrcmb.25.3.4483
  10. Iikura, M., Yamaguchi, M., Hirai, K., Miyamasu, M., Yamada, H., Nakajima, T. 2001. Regulation of surface FcepsilonRI expression on human eosinophils by IL-4 and IgE. International Archives of Allergy & Immunology 124(4): 470-477. https://doi.org/10.1159/000053782
  11. Jeong, M., Yang, J., Choi, W., Kim, J., Kim, S.J., Park, M. Chemical compositions and antioxidant activities of essential oil extracted from Neolitsea aciculata (Blume) Koidz leaves. Journal of the Korean Wood Science and Technology 45(1): 96-106. https://doi.org/10.5658/WOOD.2017.45.1.96
  12. Kang, J., Son, Y., Yim, J., Jeon, J. 2016. Estimation of carbon stock and uptake for Larix Kaempferi Lamb.. Journal of Climate Change Research 7(4): 499-506. https://doi.org/10.15531/ksccr.2016.7.4.499
  13. Kim, I.H., Shin, M.Y., Kim, Y., Chon, S. 2001. Evaluation of reproductive growth in a mature stand of Korean Pine under stimulated climatic condition. Korean Journal of Agricultural and Forest Meteorology 3(4): 185-198.
  14. Kim, S., Lee, S., Hong, C., Jang, S., Lee, S., Park, M., Choi, I. 2013. Evaluation on anti-dermatophyte effect of Larix (Kaempferi) essential oil on the morphological changes of Eermatophyte fungal hyphae. Journal of the Korean Wood Science and Technology 41(3): 247-257. https://doi.org/10.5658/WOOD.2013.41.3.247
  15. Kim, S., Lee, S., Cho, S., Hong, C., Park, M., Choi, I. 2016. Evaluation on anti-fungal activity and synergy effects of essential oil and their constituents from Abies holophylla. Journal of the Korean Wood Science and Technology 44(1): 113-123. https://doi.org/10.5658/WOOD.2016.44.1.113
  16. Kim, S., Lee, S., Cho, S., Hong, C., Park, S., Park, M., Choi, I. 2017. Antioxidant activities of Cryptomeria japonica leaves extracts by extraction methods. Journal of the Korean Wood Science and Technology 45(5): 495-510. https://doi.org/10.5658/WOOD.2017.45.5.495
  17. Kindt, T.J., Goldsby, R.A., Osborne, B.A., Kuby, J. 2007. Kuby Immunology (6th ed.), W.H. Freeman Publishers, New York, NY, USA.
  18. Kong, W. 2004. Species composition and distribution of native Korean conifers. Journal of the Korean Geographical Society 39(4): 528-543.
  19. Kwon, H., Song, C. 2012. Effect of Scutellaria barbata pharmacopuncture extract on degranulation and inflammatory mediator release in RBL-2H3 cells. Korean Journal of Acupuncture 29(3): 406-420.
  20. Lee, M.., Park, S., Lee, S., Lee, H., Kil, D. 2014. Emission characteristics of volatile organic compounds by humidifier with using Hinoki cypress extracts. Journal of the Korean Wood Science and Technology 42(6): 747-757. https://doi.org/10.5658/WOOD.2014.42.6.747
  21. Li, Z., Chen, S., Yang, X., Wang, R., Min, H., Wu, L., Si, C., Bae, Y. 2018. Secondary metabolites with anti-complementary activity from the stem barks of Juglans mandshurica Maxim. Journal of the Korean Wood Science and Technology 46(2): 118-124. https://doi.org/10.5658/WOOD.2018.46.2.118
  22. Mastusa, H., Morikawa, T., Ueda, K., Managi, H., Yoshikawa, M. 2002. Structural requirements of flavonoids for inhibition of antigen-induced degranulation, TNF-${\alpha}$ and IL-4 production from RBL-2H3 cells. Bioorganic & Medicinal Chemistry 10(10): 3123-3128. https://doi.org/10.1016/S0968-0896(02)00227-4
  23. Min, H., Kim, Y., Bae, Y. 2017. Evaluation of biological activity on hawthorn tree (Crataegus pinnatifida) extracts. Journal of the Korean Wood Science and Technology 45(3): 317-326. https://doi.org/10.5658/WOOD.2017.45.3.317
  24. Prussin, C., Metcalfe, D.D. 2003. 4. IgE, mast cells, basophils, and eosinophils. Journal of Allergy and Clinical Immunology 111(2): S486-494. https://doi.org/10.1067/mai.2003.120
  25. Rho, J., Shin, E., Shin, S. 2014. Antifungal activities of essential oil from the roots of Angelica dahurica Bentham et Hooker f.. Yakhak Hoeji 58(1): 58-61.
  26. Rufino, A.T., Ribeiro, M., Judas, F., Salgueiro, L., Lopes, M.C., Cavaleiro, C., Mendes, A.F. 2014. Antiinflammatory and chondroprotective activity of (+)-${\alpha}$-pinene: structural and enantiomeric selectivity. Journal of Natural Products 77(2): 264-269. https://doi.org/10.1021/np400828x
  27. Rufino, A.T., Ribeiro, M., Sousa, C., Judas, F., Salgueiro, L., Cavaleiro, C., Mendes, A.F. 2015. Evaluation of the anti-inflammatory, anti-catabolic and pro-anabolic effects of e-caryophyllene, myrcene and limonene in a cell model of osteoarthritis. European Journal of Pharmacology 750: 141-150. https://doi.org/10.1016/j.ejphar.2015.01.018
  28. Son, K., Song, J., Um, S., Paek, K.Y., Oh, H., Lee, J., Kim, J. 2001. Effect of absorption of essential oils on the changes of arousal and antistress. Horticulture Environment and Biotechnology 42(5): 614-620.
  29. Tumen, I., Akkol, E.K., Süntar, I., Keles, H. 2011. Wound repair and anti-inflammatory potential of essential oils from cones of Pinaceae: preclinical experimental research in animal models. Journal of Ethnopharmacology 137(3): 1215-1220. https://doi.org/10.1016/j.jep.2011.07.046
  30. Yang, J., Ahn, C., Jeung, E., Choi, W., Kim, J., Park, M. 2017. Anti-inflammatory activity of essential oil extracted from Chamaecyparis obtusa (Sieb. et Zucc.) wood. Korean Journal of Pharmacognosy 48(1): 18-24.
  31. Yun, M.S., Cho, H.M., Yeon, B.R., Choi, J.S., Kim, S.M. 2013. Herbicidal activities of essential oils from pine, pine nut, larch and khigan fir in Korea. Weed and Turfgrass Science 2(1): 30-37. https://doi.org/10.5660/WTS.2013.2.1.030
  32. Yoon, S.J., Pyo, M.Y. 2012. Down-regulation of T helper 2-associated cytokine expression by fisetin. Yakhak Hoeji 56(5): 326-332.