Chlorella dichloromethane extract ameliorates NO production and iNOS expression through the down-regulation of NF kappa B activity mediated by suppressed oxidative stress in RAW 264.7 macrophages.
BACKGROUND
It has been proposed that chlorella extracts have antioxidative and anti-inflammatory effects.


METHODS
RAW 264.7 murine macrophage cell line was preincubated with various concentrations (0-100 mug/ml) of chlorella dichloromethane extract (CDE) and stimulated with lipopolysaccharide (LPS) to induce oxidative stress and inflammation.


RESULTS
Treatments of CDE reduced thiobarbituric acid-reactive substances (TBARS) accumulation, enhancing glutathione level and activities of antioxidative enzymes including superoxide dismutase (SOD), catalase, glutathione peroxidase (GSH-px), and glutathione reductase in LPS-stimulated macrophages than LPS-only treated cells. Nitric oxide (NO) production was significantly suppressed in a dose-dependent manner (p<0.05) with an IC(50) of 30.5 microg/ml. Treatment of CDE at 50 microg/ml suppressed NO production to 6% of LPS-control. Treatment with CDE suppressed the levels of inducible nitric oxide synthase (iNOS) protein and mRNA expressions. The specific DNA binding activities of nuclear factor kappa B (NF kappa B) on nuclear extracts from CDE treatments were significantly suppressed with an IC(50) of 62.7 mug/ml in a dose-dependent manner.


CONCLUSIONS
CDE ameliorates NO production and iNOS expression through the down-regulation of NF kappa B activity, which may be mediated by attenuated oxidative stress in RAW 264.7 macrophages.
Park
Ji-Young
JY
School of Food and Life Science, Biohealth Product Research Center and Food Science Institute, Inje University, 607 Obang-dong, Kimhae 621-749, South Korea.
Cho
Hye-Yeon
HY
Kim
Jong-Kyung
JK
Noh
Kyung-Hee
KH
Yang
Jeong-Rye
JR
Ahn
Jeong-Mi
JM
Lee
Mi-Ok
MO
Song
Young-Sun
YS
D000818:Animals; D015153:Blotting, Western; D002374:Catalase / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002708:Chlorella / Q000737:chemistry*; D015536:Down-Regulation / Q000187:drug effects; D024202:Electrophoretic Mobility Shift Assay; D005979:Glutathione Peroxidase / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D008752:Methylene Chloride; D051379:Mice; D016328:NF-kappa B / Q000096:biosynthesis*; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000096:biosynthesis*; D052247:Nitric Oxide Synthase Type II; D018384:Oxidative Stress / Q000502:physiology*; D010088:Oxidoreductases / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012997:Solvents; D013482:Superoxide Dismutase / Q000378:metabolism; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism
D008070:Lipopolysaccharides; D016328:NF-kappa B; D012333:RNA, Messenger; D012997:Solvents; D017392:Thiobarbituric Acid Reactive Substances; D009569:Nitric Oxide; D008752:Methylene Chloride; D010088:Oxidoreductases; D002374:Catalase; D005979:Glutathione Peroxidase; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; D013482:Superoxide Dismutase; C020667:glutathione oxidase