생명과학회지 (Journal of Life Science)

  • 제28권3호
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  • Pages.293-299
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  • 2018
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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HT-1080 세포에서 만형자 용매 추출물의 암전이 억제효과

Anti-invasive Effect of the Solvent-partitioned Fractions from Viticis Fructus in PMA-induced HT-1080 Cells

  • 손재민 (한국해양대학교 해양과학기술전문대학원 해양과학기술융합학과) ;
  • 김호준 (한국해양대학교 해양과학기술대학 해양환경.생명과학부) ;
  • 공창숙 (신라대학교 의생명과학대학 식품영양학과) ;
  • 서영완 (한국해양대학교 해양과학기술전문대학원 해양과학기술융합학과)
  • Son, Jaemin (Ocean Science & Technology School, Korea Maritime & Ocean University) ;
  • Kim, Hojun (Division of Marine Bioscience, Korea Maritime & Ocean University) ;
  • Kong, Chang-Suk (Department of Food and Nutrition, College of Medical and Life Sciences, Silla University) ;
  • Seo, Youngwan (Ocean Science & Technology School, Korea Maritime & Ocean University)
  • 투고 : 2018.01.08
  • 심사 : 2018.03.21
  • 발행 : 2018.03.30
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초록

본 연구는 PMA (Phorbol 12-myristate 13-acetate)에 의해 MMP 활성이 증가된 섬유육종세포에서 MMP-2와 -9의 mRNA 발현 및 단백질 활성에 대한 만형자의 억제 효과를 zymography와 RT-PCR 방법에 의해서 측정하였다. 만형자 시료는 dichloromethane에 의해서 두 번 추출되었으며 동일한 과정을 methanol를 사용하여 반복하였다. 각각의 용매에 의해서 얻어진 추출물을 합한 후에 zymography를 이용하여 이 추출물의 MMP-2와 -9에 대한 억제효과를 측정한 결과 유의적인 억제효과를 나타내었다. 억제활성 성분을 추적하기 위하여 극성에 따른 추출물의 용매분획을 실시하여 n-hexane, 85% aq. MeOH, n-butanol, 및 water 분획층을 얻었다. 이 4가지 용매분획에 대한 MMP 억제활성을 측정하였으며 측정한 결과 85% aq. MeOH 분획층이 zymography와 RT-PCR 실험에서 MMP-2와 -9에 대해 가장 강한 억제효과를 나타내었다. 이상의 결과는 만형자 추출물이 암전이 억제제 개발을 위한 좋은 원천이 될 수 있는 가능성이 있음을 제시한다.

Viticis fructus (fruits of Vitex rotundifolia) is the dried fruit from Vitex rotundifolia; is a traditional medicine for treating inflammation, migraines, chronic bronchitis, headaches, eye pain, and gastrointestinal infections; and demonstrates various bioactivities, including anti-allergic, anti-cancer, and anti-inflammatory effects, which are partly due to its phenolic compound content. This study examines the inhibitory effects of viticis fructus (fruits of Vitex rotundifolia) on MMP-2 and MMP-9 expression using gelatin zymography and RT-PCR in phorbol-12-myristate-13-acetate (PMA)-induced HT-1080 fibro-sarcoma cells. Fruits of Vitex rotundifolia were extracted twice using dichloromethane ($CH_2Cl_2$) and methanol (MeOH). The combined crude extracts ($CH_2Cl_2$ and MeOH) significantly inhibited MMP-2 and MMP-9 activities in gelatin zymography. The combined extracts were fractionated into n-hexane, 85% aqueous methanol (85% aq. MeOH), n-butanol, and water, successively according to polarity. Among all solvent-partitioned fractions, 85% aq. MeOH fractions showed the strongest inhibition on the activation of MMP-2 and MMP-9 in gelatin zymography. In PMA-stimulated HT-1080 cells, the expression levels of MMP-2 and MMP-9 mRNA were also greatly inhibited by the 85% aq. MeOH fraction. These results suggest that viticis fructus can be used as an excellent source for anti-invasive agents.

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