Korean journal of applied entomology (한국응용곤충학회지)

Korean Society of Applied Entomology (한국응용곤충학회)
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Inhibition of Nitric Oxide Production by ladybug extracts(Harmonia axyridis) in LPS-activated BV-2 cells

무당벌레(Harmonia axyridis) 추출물에 의한 BV-2 세포주의 Nitric Oxide 생성 저해 활성

  • Han Sang-Mi (Department of Agricultural Biology, National Institute of Agricultural Science and Technology, RDA) ;
  • Lee Sang-Han (Department of Food Science and Technology, Kyungpook National University) ;
  • Yun Chi-Young (Department of Biology, Daejeon University) ;
  • Kang Seok-Woo (Department of Agricultural Biology, National Institute of Agricultural Science and Technology, RDA) ;
  • Lee Kyung-Gill (Department of Agricultural Biology, National Institute of Agricultural Science and Technology, RDA) ;
  • Kim Ik-Soo (Department of Agricultural Biology, National Institute of Agricultural Science and Technology, RDA) ;
  • Yun Eun-Young (Department of Agricultural Biology, National Institute of Agricultural Science and Technology, RDA) ;
  • Lee Pyeong-Jae (Department of Agricultural Biology, National Institute of Agricultural Science and Technology, RDA) ;
  • Kim Sun-Yeou (Graduate School of East-West Medical Science, Kyung Hee University) ;
  • Hwang Jae-Sam (Department of Agricultural Biology, National Institute of Agricultural Science and Technology, RDA)
  • 한상미 (농업과학기술원 농업생물부) ;
  • 이상한 (경북대학교 식품공학과) ;
  • 윤치영 (대전대학교 생물학과) ;
  • 강석우 (농업과학기술원 농업생물부) ;
  • 이광길 (농업과학기술원 농업생물부) ;
  • 김익수 (농업과학기술원 농업생물부) ;
  • 윤은영 (농업과학기술원 농업생물부) ;
  • 이평재 (농업과학기술원 농업생물부) ;
  • 김선여 (경희대학교 동서의학대학원) ;
  • 황재삼 (농업과학기술원 농업생물부)
  • Published : 2006.04.01
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Abstract

Inflammation in the brain has known to be associated with the development of a various neurologiacal diseases. The hallmark of neuro-inflammation is the activation of microglia, brain macrophage. Pro-inflammatory compounds including nitric oxide(NO) are the main cause of neuro-degenerative disease such as Alzheimer's disease. In the study, we examined whether Harmonia axyridis extracts inhibit the NO production by a direct method using Griess reagent, western blotting and by RT-PCR(Reverse Transcription-Polymerase Chain Reactionin) the gene expression of inducible nitric oxide synthase(iNOS). Distilled water$(H_2O)$ and methanol(MeOH) extracts of H. axyridis inhibited the protein expression of TNF-a(Tumor Necrosis Factor) and IL-6(Interleukin) in LPS (Lipopolysaccharide) stimulated BV-2 cells at the concentration of 100 ng/ml. Incubation of BV-2 cells with the extracts of $H_2O$ of MeOH inhibited the LPS induced NO and iNOS protein. And this inhibition of iNOS protein is concordant with the inhibition of iNOS mRNA expression. These data suggested that H. axyridis extracts may play a crucial role in inhibiting the NO production.

퇴행성 뇌질환은 뇌에 존재하는 면역세포인 소교세포의 염증반응이 발병 요인 중의 하나로 알려져 있다. 이에 본 연구는 초고속자동화시스템(high throughput screening: HTS)을 이용하여 약용곤충추출물로부터 항산화와 항염증 기능이 있다고 알려진 무당벌레 추출물로부터 염증발생인자인 nitric oxide의 생성에 어떠한 영향을 주는지 다음과 같은 결과를 얻었다. 소교세포인 BV-2세포에 대한 무당벌레 추출물의 세포독성은 물과 메탄올, DMSO 추출물에서는 100 ng/ml 까지는 거의 없었으나 에탄올 추출물은 1 ng/ml에서도 세포독성이 있었다. 물과 메탄을 추출물(50 ng/ml)은 LPS로 활성화된 BV-2세포에서 $TNF-{\alpha}$와 $IL-1{\beta}$의 발생을 35-60% 가량 억제 하였다. LPS로 유도된 NO의 생성은 물과 메탄올 추출물을 처리했을 때 각각 55%, 76% 억제되었다. 또한, MeOH 추출물을 처리했을 경우 LPS에 의한 iNOS 발현 정도를 단백질 수준과 mRNA 수준에서 현저하게 억제시킴을 확인하였다.

Keywords

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