Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Isolation of Anti-inflammatory Active Substance β-Sitosterol from Seabuckthorn (Hippophae rhamnoides L.) Stem

비타민나무(Hippophae rhamnoides L.) 줄기로부터 항염 활성물질 β-Sitosterol의 분리

  • Park, Yu-Hwa (Gangwon Provincial Agricultural Research & Extention Services) ;
  • Lim, Sang-Hyun (Gangwon Provincial Agricultural Research & Extention Services) ;
  • Ham, Hun-Ju (Gangwon Provincial Agricultural Research & Extention Services) ;
  • Kim, Hee-Yeon (Gangwon Provincial Agricultural Research & Extention Services) ;
  • Jeong, Haet-Nim (Ginseng & Medicinal Plants Experiment Station) ;
  • Kim, Kyung-Hee (Gangwon Provincial Agricultural Research & Extention Services) ;
  • Kim, Song-Mun (Dept. of Biological Environment, Kangwon National University)
  • 박유화 (강원도 농업기술원 농산물이용시험장) ;
  • 임상현 (강원도 농업기술원 농산물이용시험장) ;
  • 함헌주 (강원도 농업기술원 농산물이용시험장) ;
  • 김희연 (강원도 농업기술원 농산물이용시험장) ;
  • 정햇님 (강원도 농업기술원 인삼약초시험장) ;
  • 김경희 (강원도 농업기술원 농산물이용시험장) ;
  • 김성문 (강원대학교 바이오자원환경학과)
  • Received : 2010.03.30
  • Accepted : 2010.05.17
  • Published : 2010.07.31
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Abstract

The objective of this study was to isolate and identify anti-inflammatory chemicals in Hippophae rhamnoides L. which was grown in Chuncheon, Korea. Treatment of ethanol extracts from stems, leaves, roots, and fruits to RAW 264.7 cells reduced amounts of nitrite by 56.0, 31.9, 49.1, and 18.9% respectively, compared to only lipopolysaccharide (LPS) treatment which is well-known as a inflammation-inducing agent. The stems were extracted with hexane, dichloromethane, ethyl acetate, butanol, and water and their nitrite contents in RAW 264.7 cells were measured. The dichloromethane extracts showed the highest inflammatory activity, exhibiting 80% reduction of the nitrite content at 1 mg/mL treatment. Activity-directed fractionation of dichloromethane extracts led to the identification of $\beta$-sitosterol as the anti-inflammatory chemical. 0.1 mg/mL treatment of $\beta$-sitosterol inhibited strongly the production of nitrite by 65%, compared to only LPS treatment. These results suggest that stem of H. rhamnoides L. may be useful for inflammation treatment.

이번 연구는 강원도 춘천에서 재배된 비타민나무의(Hippophae rhamnoides L.) 부위별 추출물을 대상으로 항염활성 작용에 미치는 영향을 구명하였고, 이에 함유되어 있는 항염활성물질을 fraction directed isolation 방법으로 단리하였다. 항염활성 측정을 위하여 비타민나무의 줄기, 잎, 뿌리, 열매의 에탄올 추출물에 대하여 nitric oxide(NO) 생성량 측정 결과 염증 유발 물질인 lipopolysaccharide(LPS) 대비 각각 56.0, 31.9, 49.1, 18.9%의 저해 활성을 나타내었다. 비타민나무 줄기를 다시 극성을 달리하여 용매로 분획을 실시하였고 각각의 분획층에 대하여 NO 생성량 측정 결과 $CH_2Cl_2$ 분획층(1 mg/mL)에서 염증유발물질인 LPS 단독 처리군 대비 80%의 높은 항염활성을 나타내었다. 이에 항염활성이 가장 높았던 dichloromethane 분획층을 MCA-MCK까지 총 11개 분획층으로 분리하였고, 이 11개의 분획층에 대하여 NO 생성량을 측정한 결과 MCD 분획층이 0.1 mg/mL 농도에서 LPS 대비 65%의 가장 높은 항염활성을 나타내었다. MCD의 구조를 밝히기 위하여 GC/MS, $^{13}C$-NMR, $^1H$-NMR 분석을 수행한 결과, 항염활성물질 MCD는 분자량 414.71 g/mol, 화학구조식이 $C_{29}H_{50}O$인 $\beta$-sitosterol로 결정하였다. 향후 비타민나무 줄기는 염증예방 효능을 지닐 수 있는 기능성식품 및 의약품의 원료, 가공품 등 다양하게 활용될 수 있을 것이라 사료된다.

Keywords

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