The Protective Effect of Chlorophyll a Against Oxidative Stress and Inflammatory Processes in LPS-stimulated Macrophages

  • Park, Ji-Young (BK21 Center of Smart Food and Drug, Biohealth Products Research Center, Inje University) ;
  • Park, Chung-Mu (BK21 Center of Smart Food and Drug, Biohealth Products Research Center, Inje University) ;
  • Kim, Jin-Ju (BK21 Center of Smart Food and Drug, Biohealth Products Research Center, Inje University) ;
  • Noh, Kyung-Hee (BK21 Center of Smart Food and Drug, Biohealth Products Research Center, Inje University) ;
  • Cho, Chung-Won (School of Biomedical and Biotechnology, Inje University) ;
  • Song, Young-Sun (BK21 Center of Smart Food and Drug, Biohealth Products Research Center, Inje University)
  • Published : 2007.04.30

Abstract

This study was designed to investigate the suppressive effect of chlorophyll a on nitric oxide (NO) production and intracellular oxidative stress. In addition, chlorophyll a regulation of nuclear factor (NF) ${\kappa}B$ activation and inducible NO synthase (iNOS) expression were explored as potential mechanisms of NO suppression in a lipopolysaccharide (LPS)-stimulated macrophage cell line. RAW 264.7 murine macrophages were preincubated with various concentrations ($0-10\;{\mu}g/ mL$) of chlorophyll a and stimulated with LPS to induce oxidative stress and inflammatory response. Treatment with chlorophyll a reduced the accumulation of thiobarbituric acid-reactive substances (TBARS), enhancing glutathione level and the activities of antioxidative enzymes including superoxide dismutase, catalase, glutathione peroxidase (GSH-px), and glutathione reductase in LPS-stimulated macrophages compared to LPS-only treated cells. NO production was significantly suppressed in a dose-dependent manner (p<0.05) with an $IC_{50}$ of $12.8\;{\mu}g/mL$. Treatment with chlorophyll a suppressed the levels of iNOS protein and its mRNA expression. The specific DNA binding activities of NFkB on nuclear extracts from chlorophyll a treated cells were significantly suppressed in a dose-dependent manner with an $IC_{50}$ of $10.7\;{\mu}g/mL$. Chlorophyll a ameliorates NO production and iNOS expression through the down-regulation of NFkB activity, which may be mediated by attenuated oxidative stress in RAW 264.7 macrophages.

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