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Inhibitory Effect of Extracts of Platycodon grandiflorum (the Ballon Flower) on Oxidation and Nitric Oxide Production

도라지 부탄올 추출물의 항산화 및 nitric oxide 생성 저해 효과

  • Jang, Joo-Ri (Division of Marine Environment & Bioscience, Korea Maritime University) ;
  • Hwang, Seong-Yeon (Division of Marine Environment & Bioscience, Korea Maritime University) ;
  • Lim, Sun-Young (Division of Marine Environment & Bioscience, Korea Maritime University)
  • 장주리 (한국해양대학교 해양환경생명과학부) ;
  • 황성연 (한국해양대학교 해양환경생명과학부) ;
  • 임선영 (한국해양대학교 해양환경생명과학부)
  • Received : 2010.08.10
  • Accepted : 2010.12.17
  • Published : 2011.02.28

Abstract

We explored the effect of extracts of dried Platycodon grandiflorum on production of reactive oxygen species (ROS), glutathione (GSH) and nitric oxide (NO). To determine antioxidant activity in the presence of $H_2O_2$-induced oxidative stress, DCFH-DA (dichlorodihydrofluorescin diacetate) assay was employed. Acetone/methylene chloride (A+M) and methanolic (MeOH) extracts of P. grandiflorum reduced intracellular ROS levels. Of the various tested fractions, n-BuOH fraction showed the highest protective effect in terms of lipid peroxide production. Total GSH levels were measured after treatment of HT1080 cells with the A+M and MeOH extracts, and other solvent fractions, at various concentration. The A+M extacts and 85% (v/v) aqueous MeOH fraction significantly increased GSH levels (p<0.05). When lipopolysaccharide (LPS)-induced NO production was evaluated, all tested crude extracts, and fractions thereof, significantly reduced NO production (p<0.05), and the n-BuOH and 85% (v/v) aqueous MeOH fractions (at 0.05 mg/mL) showed the strongest inhibitory effects. The results showed that the n-BuOH fraction inhibited both cellular oxidation and NO production, and this fraction may thus contain valuable active compounds.

본 연구에서는 도라지 분말을 유기용매로 추출한 후 도라지의 추출물과 분획물들에 의한 세포 내 활성산소종 및 glutathione (GSH)를 측정하여 항산화효과를 검토하였고 NO 생성 저해 효과를 알아보았다. 세포 내 활성산소종 생성억제 실험에서 건조 도라지의 A+M 및 MeOH 추출물과 추출물을 n-hexane, 85% aq. MeOH, n-BuOH, water로 다시 추출하여 얻어진 각각의 분획물들을 농도별로 HT1080 세포에 처리하였을 때 A+M과 MeOH 추출물 모두 측정시간 120분 동안 $500\;{\mu}M$ $H_2O_2$만을 처리한 control군에 비해 세포 내 활성산소종을 크게 억제시켰으며, MeOH 추출물에 의한 항산화 효과가 더 높게 나타났다. 또한, 각 분획물들 중 n-BuOH 분획물이 다른 분획물들에 비해 우수한 항산화 활성을 보였다. GSH 농도 측정 실험에서 A+M 및 85% aq. MeOH 분획물를 처리했을 때 GSH 함량이 증가하였다. NO 생성 저해 실험에서는 A+M 및 MeOH 추출물이 0.01 및 0.05 mg/mL의 농도에서 NO 생성을 억제하는 것으로 나타났으며, A+M 추출물에 의한 저해 효과가 높았다. 각 분획물들은 모두 control보다 낮은 NO 생성량을 나타내었으며, 특히 85% aq. MeOH 및 n-BuOH 분획물은 0.05 mg/mL 농도에서 blank에 가까운 NO 생성 억제율을 나타냈다. 이상의 연구결과로부터 n-BuOH 분획물에 의한 세포 내 활성산소종 생성 억제 효과 및 NO 생성 저해 효과가 우수함을 알 수 있었으며, 향후 분획물의 분리 정제를 통한 새로운 기능성 물질의 개발이 필요할 것으로 사료된다.

Keywords

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