Journal of Korean Traditional Oncology (대한암한의학회지)

Korean Association of Traditional Oncology (대한암한의학회)
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Inhibition of LPS-induced Inflammatory Biomarkers by Fraction of Citrus hassaku pericarp through Suppression of NF-${\kappa}B$ Activation in RAW264.7 Cells

재래감귤 팔삭의 과피 추출물이 LPS로 활성화 된 RAW264.7 대식세포에서 염증매개물질 억제에 미치는 효과

  • Kim, Chul-Won (Department of Oriental Pathology, College of Oriental Medicine, Kyung Hee University) ;
  • Kim, Sung-Moo (Department of Oriental Pathology, College of Oriental Medicine, Kyung Hee University) ;
  • Jeong, Seung-Weon (Korea Food Research Institute) ;
  • K., So-Mi Cho (Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University) ;
  • Ahn, Kwang-Seok (Department of Oriental Pathology, College of Oriental Medicine, Kyung Hee University)
  • 김철원 (경희대학교 한의과대학 기초한의학과 병리학교실) ;
  • 김성무 (경희대학교 한의과대학 기초한의학과 병리학교실) ;
  • 정승원 (한국식품연구원) ;
  • 김소미 (제주대학교 생명공학부) ;
  • 안광석 (경희대학교 한의과대학 기초한의학과 병리학교실)
  • Received : 2011.12.05
  • Accepted : 2011.12.23
  • Published : 2011.12.28
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Abstract

Objectives : Citrus is the fruit that is readily available around us. Therefore, we investigated the anti-inflammatory effects of fraction isolated from the Citrus hassaku pericarp in RAW264.7 macrophage cells. Methods : The effects of fraction from Citrus hassaku pericarp on cell viability on RAW264.7 cells were measured by the MTT assay. The mRNA levels of iNOS and COX-2, its protein level by fraction of Citrus hassaku pericarp treatment in RAW264.7 macrophage cells were investigated by RT-PCR and immunoblots. Nitrite accumulation in the culture was measured colorimetrically by the Griess reaction using a Griess reagent. The amount of IL-6 and TNF-${\alpha}$ production was determined using an enzyme-linked immunosorbent assay (ELISA) kit. Results : The results indicated that the fraction of Citrus hassaku pericarp concentration highly suppressed lipopolysaccharide (LPS)-induced nitric oxide (NO) and IL-6 productions without a cytotoxic effect on RAW264.7 cells. fraction of Citrus hassaku pericarp inhibited the expressions of LPS-induced iNOS and COX-2 protein and their mRNA in a dose-dependent manner. Particularly, fraction of Citrus hassaku pericarp suppressed the level of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activity, which was linked with the suppression of LPS-induced phosphorylation of p65 at serine 276 and p65 translocation into nuclei, but not MAPK signaling. In addition, treatment with fraction of Citrus hassaku pericarp inhibited the production of IL-6 and TNF-${\alpha}$ in LPS-stimulated RAW264.7 cells. Conclusion : Our results indicate that fraction of Citrus hassaku pericarp potentially inhibits the biomarkers related to inflammation through the blocking of NF-${\kappa}B$ p65 activation, and it may be a potential therapeutic candidate for the treatment of inflammatory diseases.

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