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Anti-inflammatory Effect of Myricetin from Rhododendron mucronulatum Turcz. Flowers in Lipopolysaccharide-stimulated Raw 264.7 Cells

Lipopolysaccharide로 유도된 Raw264.7 cell에서 Rhododendron mucronulatum Turcz. Flower으로부터 분리한 myricetin에 의한 염증 억제효과

  • Choi, Moo-Young (Department of Food and Nutrition, College of Health Science, Sangji University) ;
  • Hong, Shin-Hyup (School of Food Science and Biotechnology/Food Bio-Industry Institute, College of Agriculture and Life Sciences, Kyungpook National University) ;
  • Cho, Jun-Hyo (School of Food Science and Biotechnology/Food Bio-Industry Institute, College of Agriculture and Life Sciences, Kyungpook National University) ;
  • Park, Hye-Jin (School of Food Science and Biotechnology/Food Bio-Industry Institute, College of Agriculture and Life Sciences, Kyungpook National University) ;
  • Jo, Jae-Bum (School of Food Science and Biotechnology/Food Bio-Industry Institute, College of Agriculture and Life Sciences, Kyungpook National University) ;
  • Lee, Jae-Eun (School of Food Science and Biotechnology/Food Bio-Industry Institute, College of Agriculture and Life Sciences, Kyungpook National University) ;
  • Kim, Dong-Hee (Korea Promotion Institute for Traditional Medicine Industry) ;
  • Kim, Byung-Oh (School of Food Science and Biotechnology/Food Bio-Industry Institute, College of Agriculture and Life Sciences, Kyungpook National University) ;
  • Cho, Young-Je (School of Food Science and Biotechnology/Food Bio-Industry Institute, College of Agriculture and Life Sciences, Kyungpook National University)
  • 최무영 (상지대학교 보건과학대학 식품영양학과) ;
  • 홍신협 (경북대학교 농업생명과학대학 식품공학부/식품생물산업연구소) ;
  • 조준효 (경북대학교 농업생명과학대학 식품공학부/식품생물산업연구소) ;
  • 박혜진 (경북대학교 농업생명과학대학 식품공학부/식품생물산업연구소) ;
  • 조재범 (경북대학교 농업생명과학대학 식품공학부/식품생물산업연구소) ;
  • 이재은 (경북대학교 농업생명과학대학 식품공학부/식품생물산업연구소) ;
  • 김동희 (한국한방산업진흥원) ;
  • 김병오 (경북대학교 농업생명과학대학 식품공학부/식품생물산업연구소) ;
  • 조영제 (경북대학교 농업생명과학대학 식품공학부/식품생물산업연구소)
  • Received : 2016.06.03
  • Accepted : 2016.10.05
  • Published : 2016.11.30

Abstract

As a research of inflammation inhibitory activity using natural resource, the inflammation inhibitory activity by purified active compound from Rhododendron mucronulatum flower was experimented. Rhododendron mucronulatum flower components were purified and separated with Sephadex LH-20 and MCI gel CHP-20 column chromatography, Purified compound was confirmed as myricetin by $^1H-NMR$, $^{13}C-NMR$ and Fast atom bombardment (FAB)-Mass spectrum to have inhibition activity on inflammatory factors secreted by Raw 264.7 cells in response to lipopolysaccharide stimulation. Myricetin inhibited nitric oxide (NO) expression in a concentration dependent manner, approximately 40% inhibition was observed at a concentration of $50{\mu}M$. The inhibition effect of myricetin on inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 protein expression was 20% and 80%, respectively, at a concentration of $25{\mu}M$. Myricetin also inhibited expression of the inflammatory cytokines, tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\beta}$, IL-6 and prostaglandin $E_2(PGE_2)$ in a concentration dependent manner; a concentration of $50{\mu}M$, 70%, 80%, 80% and 95% inhibition was observed, respectively. Therefore myricetin isolated from Rhododendron mucronulatum flowers is expected to have an anti-inflammatory effect in Raw 264.7 cell induced by lipopolysaccharides. The results can be expected myricetin from Rhododendron mucronulatum flower to use as functional resource for anti-inflammatory activity.

진달래꽃으로부터 Sephadex LH-20 및 MCI gel CHP-20 column chromatography로 정제한 결과 항염증 활성을 가지는 myricetin을 분리, 동정하였다. Myricetin은 농도 의존적으로 NO 발현을 억제하였고, $50{\mu}M$ 농도에서 약 40%의 억제효과를 나타내었다. Myricetin의 iNOS와 COX-2의 발현억제력은, $25{\mu}M$ 농도에서 각각 20% 및 80%의 protein 발현 억제효과를 나타내었다. 또한 myricetin의 염증반응의 cytokine을 측정하여 $TNF-{\alpha}$, $IL-1{\beta}$, IL-6 및 $PGE_2$의 억제력을 살펴본 결과, 농도 의존적으로 발현억제 효과를 나타내었으며, $50{\mu}M$ 농도에서 각각 70%, 80%, 80% 및 95%의 발현 억제효과를 나타내었다. 따라서 진달래 꽃잎에서 분리한 myricetin은 LPS로 유도되어진 대식세포주인 Raw 264.7 세포에서 염증반응의 억제효과를 기대할 수 있었다.

Keywords

References

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