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Antiinflammatory Activity of Solvent-partitioned Fractions from Atriplex gmelinii C. A. Mey. in LPS-stimulated RAW264.7 Macrophages

염생식물 가는갯는쟁이 용매 추출물의 항염증활성

  • Jeong, Heejeong (Department of Convergence Study on the Ocean Science and Technology, Ocean Science and Technology School, Korea Maritime and Ocean University) ;
  • Kim, Hojun (Division of Marine Bioscience, College of Ocean Science and Technology, Korea Maritime and Ocean University) ;
  • Ju, Eunsin (Division of Marine Bioscience, College of Ocean Science and Technology, Korea Maritime and Ocean University) ;
  • Lee, Seul-Gi (Department of Food and Nutrition, College of Medical and Life Sciences, Silla University) ;
  • Kong, Chang-Suk (Department of Food and Nutrition, College of Medical and Life Sciences, Silla University) ;
  • Seo, Youngwan (Department of Convergence Study on the Ocean Science and Technology, Ocean Science and Technology School, Korea Maritime and Ocean University)
  • 정희정 (한국해양대학교 해양과학기술전문대학원 해양과학기술융합학과) ;
  • 김호준 (한국해양대학교 해양과학기술대학 해양환경.생명과학부) ;
  • 주은신 (한국해양대학교 해양과학기술대학 해양환경.생명과학부) ;
  • 이슬기 (신라대학교 의생명과학대학 식품영양학과) ;
  • 공창숙 (신라대학교 의생명과학대학 식품영양학과) ;
  • 서영완 (한국해양대학교 해양과학기술전문대학원 해양과학기술융합학과)
  • Received : 2016.09.27
  • Accepted : 2016.11.02
  • Published : 2017.02.28

Abstract

As a part of ongoing research to elucidate and characterize antiinflammatory nutraceuticals, the crude extracts from Atriplex gmelinii C. A. Mey. and their solvent-partitioned fractions were tested for their antiinflammatory potential in lipopolysaccharide (LPS)-stimulated RAW 264.7 mouse macrophages. The crude extracts of A. gmelinii C. A. Mey. were fractioned according to polarity with n-hexane, 85% aqueous methanol (85% aq. MeOH), n-butanol, and $H_2O$. Their antiinflammatory activities were investigated in LPS-induced inflammation in mouse macrophages by measuring nitric oxide (NO) generation and mRNA expression of inflammation mediators, namely, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-$1{\beta}$ ($IL-1{\beta}$), and IL-6. As a result, we confirmed that the crude extracts of A. gmelinii C. A. Mey. inhibited LPS-stimulated NO production and mRNA expression of iNOS and COX-2 as important inflammatory factors. The inhibition of NO production through the downregulation of important inflammatory factors such as iNOS, COX-2, $IL-1{\beta}$, and IL-6 was found by treatment with all solvent-partitioned fractions. Among all tested fractions, 85% aq. MeOH showed the strongest antiinflammatory response. Based on the current results, A. gmelinii C. A. Mey. was suggested to possess natural antiinflammatory components, indicating that it could be used as a valuable source of antiinflammatory substances.

본 연구에서는 염생식물을 이용한 생리활성 소재 개발 연구의 일환으로 가는갯능쟁이의 용매 추출물 및 분획물을 제조하여 LPS로 염증을 유도한 마우스 유래 macrophages에서의 NO 생성 및 염증관련인자의 발현에 미치는 영향을 검토하여 가는갯능쟁이의 항염증 소재로서의 이용 가능성을 검토하였다. 가는갯능쟁이는 methylene chloride와 methanol을 이용하여 제조한 조추출물(crude extract, 48.3 mg)을 용매극성에 따라 단계적으로 분획하여 n-hexane 층(11.5 mg), 85% aqueous methanol 수용액층(85% aq. MeOH, 11.3 mg), n-butanol 층(n-BuOH, 11.2 mg)과 water 층(12.3 mg)을 얻었다. 가는갯능쟁이의 조추출물 및 분획물의 RAW264.7 대식세포에 대한 독성 여부를 측정하여 세포에 대한 독성이 나타나지 않는 처리 농도에서 추출물 및 분획물의 항염증 활성을 확인하였다. 조추출물은 LPS로 유도된 NO 생성을 유의적으로 억제하는 효과를 나타내었으며, 주요 염증 유발인자인 iNOS와 COX-2의 발현 또한 효과적으로 억제하였다. 분획물의 NO 생성 억제효과는 85% aq. MeOH> n-BuOH >n-hexane > $H_2O$의 순으로 나타났으며, 세포내 전사수준에서 염증관련 유전자의 발현 억제율은 $H_2O$ 분획 처리군에 비해 85% aq. MeOH, n-BuOH 및 n-hexane 분획 처리군에서 높게 나타났다. 이상의 결과로부터 항염증 활성이 가장 높게 나타난 85% aq. MeOH 분획층으로 부터 염증억제 효과가 있는 유효 성분의 분리가 기대된다.

Keywords

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