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Anti-inflammatory Effect of Ethanol Extract from Sargassum fulvellum on Lipopolysaccharide Induced Inflammatory Responses in RAW 264.7 Cells and Mice Ears

LPS로 유도된 RAW 264.7 세포와 마우스 귀조직에 대한 참모자반 (Sargassum fulvellum) 에탄올 추출물의 항염증 효과

  • Jeong, Da-Hyun (Department of Food Science & Technology/Institute of Food Science and Pukyong National University) ;
  • Kim, Koth-Bong-Woo-Ri (Institute of Fisheries Sciences, Pukyong National University) ;
  • Kim, Min-Ji (Institute of Fisheries Sciences, Pukyong National University) ;
  • Kang, Bo-Kyeong (Department of Food Science & Technology/Institute of Food Science and Pukyong National University) ;
  • Bark, Si-Woo (Department of Food Science & Technology/Institute of Food Science and Pukyong National University) ;
  • Pak, Won-Min (Department of Food Science & Technology/Institute of Food Science and Pukyong National University) ;
  • Kim, Bo-Ram (Department of Food Science & Technology/Institute of Food Science and Pukyong National University) ;
  • Ahn, Na-Kyung (Department of Food Science & Technology/Institute of Food Science and Pukyong National University) ;
  • Choi, Yeon-Uk (Department of Food Science & Technology/Institute of Food Science and Pukyong National University) ;
  • Ahn, Dong-Hyun (Department of Food Science & Technology/Institute of Food Science and Pukyong National University)
  • 정다현 (부경대학교 식품공학과/식품연구소) ;
  • 김꽃봉우리 (부경대학교 수산과학연구소) ;
  • 김민지 (부경대학교 수산과학연구소) ;
  • 강보경 (부경대학교 식품공학과/식품연구소) ;
  • 박시우 (부경대학교 식품공학과/식품연구소) ;
  • 박원민 (부경대학교 식품공학과/식품연구소) ;
  • 김보람 (부경대학교 식품공학과/식품연구소) ;
  • 안나경 (부경대학교 식품공학과/식품연구소) ;
  • 최연욱 (부경대학교 식품공학과/식품연구소) ;
  • 안동현 (부경대학교 식품공학과/식품연구소)
  • Received : 2014.03.26
  • Accepted : 2014.05.26
  • Published : 2014.08.31

Abstract

This study investigated the anti-inflammatory effects of Sargassum fulvellum ethanol extract (SFEE) on the lipopolysaccharide (LPS)-induced inflammatory response. SFEE remarkably suppressed production of NO and pro-inflammatory cytokines (IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ at 50 and $100{\mu}g/mL$. There were no cytotoxic effects on proliferation of macrophages treated with SFEE compared to the control. SFEE reduced expression of iNOS and COX-2 proteins in a dose-dependent manner. The formation of edema in mouse ears was reduced at the highest dose tested compared to the control. Moreover, in the acute toxicity test, no mortality occurred in mice administered 5,000 mg/kg body weight of SFEE over the 2-week observation period. These results suggest that SFEE may have significant effects on inflammatory factors and be a potential anti-inflammatory therapeutic material.

본 연구에서는 천연식물자원인 참모자반 에탄올 추출물(SFEE)의 항염증 활성을 알아보기 위하여 in vitro 및 in vivo 실험을 진행하였다. 먼저 SFEE의 세포독성을 살펴보기 위하여 RAW 264.7 대식세포를 이용하여 세포 생존율을 살펴본 결과, SFEE $100{\mu}g/mL$의 농도까지 전혀 독성을 나타내지 않았다. 또한 항염증 효과를 알아보기 위하여 in vitro type으로 LPS로 염증을 유도한 RAW 264.7 세포에 대하여 NO, IL-6, $TNF-{\alpha}$$IL-1{\beta}$ 분비량과 iNOS, COX-2 및 $NF-{\kappa}B$와 같은 염증성 단백질의 발현량을 측정하였다. 대식세포 배양액의 $NO{_2}{^-}$ 농도를 측정한 결과 SFEE 첨가 농도에 따라 유의적으로 분비량이 감소하는 것을 확인하였으며, $100{\mu}g/mL$의 농도에서 90% 이상 억제됨을 확인하였다. 또한 $100{\mu}g/mL$의 농도로 첨가 시 염증성 사이토카인인 IL-6, $TNF-{\alpha}$$IL-1{\beta}$ 분비량을 각각 79, 73, 87% 억제함을 확인하였으며, NO 생성과 연관이 있는 iNOS 단백질과 COX-2 및 $NF-{\kappa}B$ 단백질의 발현이 현저히 억제됨을 확인하였다. 이러한 SFEE의 항염증 효과를 동물모델에서 확인하기 위해 in vivo 실험을 진행한 결과 croton oil로 유도한 마우스 귀 부종을 유의적으로 억제함을 확인하였고, 조직검사 결과 염증 반응에 의한 경피 및 진피 두께의 발달을 현저히 억제시키고 염증성 세포인 비만세포의 조직으로의 침윤을 억제하여 항염증 효과를 가짐을 입증하였다. 결론적으로 참모자반 에탄올 추출물은 염증 반응의 전사인자인 $NF-{\kappa}B$의 발현을 억제하여 iNOS와 COX-2의 발현을 억제시키고 그에 의해 NO, IL-6, $TNF-{\alpha}$$IL-1{\beta}$ 분비를 억제하여 항염증 활성을 가지는 것을 확인하였으며, 따라서 참모자반 에탄올 추출물이 염증성 질환의 예방 및 개선에 유용할 것으로 사료된다.

Keywords

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