Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Identification of 5-Hydroxy-3,6,7,8,3',4'-Hexamethoxyflavone from Hizikia fusiforme Involved in the Induction of the Apoptosis Mediators in Human AGS Carcinoma Cells

  • Kim, Min Jeong (Department of Biotechnology, Dong-A University) ;
  • Lee, Hye Hyeon (Department of Biotechnology, Dong-A University) ;
  • Seo, Min Jeong (Medi-Farm Industrialization Research Center, Dong-A University) ;
  • Kang, Byoung Won (Medi-Farm Industrialization Research Center, Dong-A University) ;
  • Park, Jeong Uck (Medi-Farm Industrialization Research Center, Dong-A University) ;
  • Kim, Kyoung-Sook (Medi-Farm Industrialization Research Center, Dong-A University) ;
  • Kim, Gi-Young (Laboratory of Immunobiology, Department of Marine Life Sciences, Jeju National University) ;
  • Joo, Woo Hong (Department of Biology, Changwon National University) ;
  • Choi, Yung Hyun (Department of Biochemistry, College of Oriental Medicine, Dong-Eui University) ;
  • Cho, Young-Su (Department of Biotechnology, Dong-A University) ;
  • Jeong, Yong Kee (Department of Biotechnology, Dong-A University)
  • Received : 2012.08.10
  • Accepted : 2012.09.18
  • Published : 2012.12.28
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Abstract

An 80% ethanol extract of Hizikia fusiforme was obtained and followed by successive fractionation using the organic solvents n-hexane, ethyl acetate, and n-butanol to identify the antioxidative substance. The aqueous part of the nbutanol fractionation step, showing high antioxidative activity, was subjected to reverse-phase liquid chromatography. As a result, a substance purified from a BB-2 fraction showed high antioxidative activity. The m/z 419 [M+H] molecular ion peak in the fraction was observed by the analysis of the ESI-LC/MS spectrum. By the analysis of 1H NMR (500 MHz, DMSO-$d_6$) and $^{13}C$ NMR (125 MHz, DMSO-$d_6$) spectra, a unique compound of the fraction was biochemically identified as a 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone (5HHMF). We also investigated the effect of 5HHMF on human gastric AGS carcinoma cells. Western blot analysis suggested that the flavone substantially increased the levels of the death receptor-associated apoptosis mediators Fas, Fas L, FADD, TRADD, and DR4 in a concentration-dependent manner. The levels of Fas, Fas L, TRADD, and DR4 in the cells treated with 5HHMF ($5{\mu}g/ml$) were approximately 26.4-, 12.8-, 6.7-, and 9.8-times higher than those of non-treated cells, respectively. Of note, the level of FADD protein in the cells exposed to 5HHMF ($1{\mu}g/ml$) increased approximately 9.6-times. In addition, the cleavage of caspase-3, -8, and -9 in cultured AGS cells treated with 5HHMF was significantly confirmed. Therefore, our results suggest that 5HHMF from H. fusiforme is involved in the induction of death receptor-associated apoptosis mediators in human gastric AGS carcinoma cells.

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