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Suppression of oxidative stress and pro-inflammatory mediators by Cymbopogon citratus D. Stapf extract in lipopolysaccharide stimulated murine alveolar macrophages
Authors:M Tiwari  UN Dwivedi  P Kakkar
Institution:1. Herbal Research Section, Indian Institute of Toxicology Research, Mahatma Gandhi Marg, P.O. Box No. 80, Lucknow 226 001, U.P, India;2. Department of Biochemistry, University of Lucknow, Lucknow, U.P., India
Abstract:Exploration of antioxidants of plant origin and their scientific validation for their immense pharmacological potential is emerging as an issue of intense research now-a-days.The effect of Cymbopogon citratus extract was seen on cell viability, oxidative stress markers i.e. ROS production, SOD activity, lipid peroxidation and GSH content of murine alveolar macrophages stressed with lipopolysaccharide. Modulation in release of NO and pro-inflammatory cytokine TNF-α along with alterations in mitochondrial membrane potential under stress were compared with known plant derived antioxidant quercetin. The extract was not found to be cytotoxic at any of the selected doses. At 5 and 10 μg the extract showed significant increase in SOD activity, GSH content (p < 0.001), decrease in ROS production as seen by fluorescent dye DCFH-DA and also MDA formation (lipid peroxidation marker) significantly. The extract also showed reduction in the release of pro-inflammatory mediators TNF-α and NO significantly indicating an anti-inflammatory effect. The extract was able to restore mitochondrial membrane potential as estimated by spectrofluorimetry using the fluorescent dye Rhodamine 123. The results suggest potential use of the cytoprotective, antioxidant and anti-inflammatory property of C. citratus in the form of dietary component and also in formulations against lung inflammatory diseases where oxidative stress plays an important role.
Keywords:AMs  alveolar macrophages  AP1  activator protein 1  ARDS  acute respiratory distress syndrome  BALF  broncho alveolar lavage fluid  BHT  butylated hydroxy toluene  COPD  chronic obstructive pulmonary disease  DCFH-DA  2&prime    7&prime  -dichlorofluoresceindiacetate  DTNB  5  5&prime  -dithiobis-(2-nitrobenzoic acid)  ELISA  enzyme-linked immunosorbent assay  FBS  fetal bovine serum  GSH  glutathione reduced  HPLC  high performance liquid chromatography  HRP  horse radish peroxidase  iNOS  inducible nitric oxide synthase  IPF  idiopathic pulmonary fibrosis  LPS  lipopolysaccharide  MDA  malondialdehyde  MMP  mitochondrial membrane potential  MTT  (3-[4  5 dimethylthiazol-2-yl]-2  5-diphenyltetrazolium bromide)  NADH  nicotinamide adenine dinucleotide  NBT  nitro blue tetrazolium  NF κB  nuclear factor kappa B  NO  nitric oxide  PBS  phosphate buffer saline  PMS  phenazine methosulfate  ROS  reactive oxygen species  RNS  reactive nitrogen species  SDS  sodium dodecyl sulfate  SOD  superoxide dismutase  TBA  thio barbituric acid  TNF  tumour necrosis factor
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