title: Evidence for Serpentine as a novel antioxidant by a redox sensitive HABP1 overexpressing cell line by inhibiting its nuclear translocation of NF-κB.
abstract: Herbal antioxidants are gradually gaining importance as dietary supplements considering the growing implications of oxidative stress in most degenerative diseases and aging. Thus, continuous attempts are made to search for novel herbal molecules with antioxidative properties, using chemical methods predominantly with the need arising for cell based assays. We have generated a stable cell line F-HABP07, by constitutively overexpressing human Hyaluronan Binding Protein1 (HABP1) in murine fibroblasts which accumulates in the mitochondria leading to excess ROS generation without any external stimuli. In the present study, we demonstrated the nuclear translocation of p65 subunit of NF-κB in F-HABP07 cells, an important signature of ROS induced signalling cascade providing us an opportunity to use it as a screening system for ROS scavengers. Using known antioxidants on our designer cell line, we have demonstrated a dose dependant reduction in ROS generation and observed inhibition of p65 subunit of NF-κB nuclear translocation, increase in glutathione content and down-regulation of apoptotic marker Bax establishing its antioxidant biosensing capacity. With the help of this cell line, we for the first time demonstrated serpentine, one of the active components from the roots of Rauwolfia serpentina (a traditional medicinal plant), to be a novel non-cytotoxic antioxidant. The authenticity of this cell line screening system based discovery was validated using standard chemical assays thus, opening up new therapeutic avenues for this herbal compound and the use of this designer cell line.
authors.affiliation: Environmental Toxicology and Biochemistry Laboratory, School of Environmental Sciences, Jawaharlal Nehru University, New Delhi-110067, India.
authors.forename: Shubhra
authors.initials: S
authors.lastname: Dutta
authors.forename: Anindya Roy
authors.initials: AR
authors.lastname: Chowdhury
authors.forename: S K
authors.initials: SK
authors.lastname: Srivastava
authors.forename: Ilora
authors.initials: I
authors.lastname: Ghosh
authors.forename: Kasturi
authors.initials: K
authors.lastname: Datta
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D015374:Biosensing Techniques; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D030008:Catharanthus / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D005347:Fibroblasts / Q000166:cytology; D016166:Free Radical Scavengers / Q000032:analysis; D015870:Gene Expression / Q000187:drug effects; D005978:Glutathione / Q000032:analysis; D006801:Humans; D051379:Mice; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism*; D010084:Oxidation-Reduction / Q000187:drug effects; D010936:Plant Extracts / Q000378:metabolism / Q000494:pharmacology; D011926:Rauwolfia / Q000737:chemistry* / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D046948:Secologanin Tryptamine Alkaloids / Q000737:chemistry / Q000494:pharmacology*; D015398:Signal Transduction; D051996:Transcription Factor RelA / Q000235:genetics / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000032:analysis
chemical_list: D000975:Antioxidants; C087299:C1QBP protein, human; D002352:Carrier Proteins; D016166:Free Radical Scavengers; D024101:Mitochondrial Proteins; D010936:Plant Extracts; D017382:Reactive Oxygen Species; C498144:Rela protein, mouse; D046948:Secologanin Tryptamine Alkaloids; D051996:Transcription Factor RelA; D051028:bcl-2-Associated X Protein; C009244:serpentine (alkaloid); D005978:Glutathione