Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Anti-inflammatory Activities of Taxifolin from Opuntia humifusa in Lipopolysaccharide Stimulated RAW 264.7 Murine Macrophages

천년초의 Taxifolin이 마우스대식세포(RAW 264.7 cell)에 미치는 항염증활성

  • Received : 2015.05.28
  • Accepted : 2015.06.26
  • Published : 2015.09.30
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Abstract

This study was performed to investigate the antiinflammatory activities of taxifolin from Opuntia humifusa. A potent anti-oxidant activity was shown from the leaf extract at $IC_{50}$ value of $38.33{\pm}1.07{\mu}g/mL$ and fruit extract at $IC_{50}$ value of $40.23{\pm}2.21{\mu}g/mL$ by 1,1-diphenyl-2-picrylhydrazyl assay. Fraction of taxifolin from leaf extract identified using high performance liquid chromatography and gas chromatography/mass spectrometry. The results of cell viability indicated that taxifolin did not show cytotoxicity on RAW 264.7 cells at $500{\mu}M$ of concentration. The result showed that taxifolin inhibited lipopolysaccharide (LPS)-induced production of Nitrite oxide. In addition, taxifolin inhibited LPS-induced tumor necrosis factor-${\alpha}$ and interleukin-6 production by cytokine assay and cyclooxygenase-2 expression by western blot analysis, meaning taxifolin has a significant anti-inflammatory effect. Our results suggested that taxifolin from Opuntia humifusa showed anti-inflammatory activities.

본 연구는 항산화 효과가 뛰어난 천년초를 이용하여 taxifolin을 순수분리하고 항염증 효과를 연구하였다. 천년초는 잎과 열매로 나누어 추출하였고 추출공정으로는 먼저 메탄올을 이용하여 조추출물을 추출하였다. 이렇게 얻어진 조추출물의 항산화 효과를 측정하였는데. 잎 추출물에서 $38.33{\pm}1.07{\mu}g/mL$로 높은 항산화 활성을 나타내었다. high performance liquid chromatography를 이용하여 taxifolin을 분리하고 확인하였다. 이렇게 얻어진 taxifolin을 이용하여 항염증 효과를 측정하였다. Taxifolin의 농도별 세포독성 측정에서는 $500{\mu}M$ 농도까지 처리 하였을 때 특별한 세포독성을 나타내지는 않았다. lipopolysaccharide에 의해 유도된 RAW 264.7 cell을 이용한 taxifolin의 염증성 cytokine 측정실험에서는 IL-6의 발현양을 $500{\mu}M$ 처리시 $2139.75{\pm}1.64pg/mL$로 대조군에 비하여 47%까지 저해하였다. 하지만 $TNF-{\alpha}$의 발현량은 대조군에 비하여 5%만 감소하였다. 염증성지표의 NO의 생성량은 $500{\mu}M$ 처리시 $17.43{\pm}0.27{\mu}M$로 대조군에 비하여 57%까지 감소 시켰으며, 이와 관련된 COX-2 단백질의 발현양은 약 66% 이상 감소시켰다. 따라서 천년초에서 추출한 taxifolin은 항염증 효과가 있음을 확인할 수 있다.

Keywords

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