title: PASS-predicted Vitex negundo activity: antioxidant and antiproliferative properties on human hepatoma cells--an in vitro study.
abstract: BACKGROUND
  Hepatocellular carcinoma is a common type of tumour worldwide with a high mortality rate and with low response to current cytotoxic and chemotherapeutic drugs. The prediction of activity spectra for the substances (PASS) software, which predicted that more than 300 pharmacological effects, biological and biochemical mechanisms based on the structural formula of the substance was efficiently used in this study to reveal new multitalented actions for Vitex negundo (VN) constituents.
  
  
  METHODS
  Experimental studies based on antioxidant and antiproliferative assays verified the predictions obtained by the PASS-predicted design strategy. Antioxidant activity of VN extract was studied using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and Ferric reducing or antioxidant power (FRAP) assays. The antiproliferative activity of VN extract against WRL68 and HepG2 was investigated based on methylthiazol tetrazolium (MTT) spectrophotometric assay.
  
  
  RESULTS
  VN extract showed 79.43% inhibition of DPPH stable radical with IC50 13.31 ± 0.18 μg/ml. This inhibition was too closed to butylated hydroxyl toluene (BHT) 82.53% (IC5013.8 ± 0.14) and gallic acid 89.51% (IC50 3.1 ± 0.08). VN extract exhibited the strongest free radical scavenging power compared with two commercial antioxidants, BHT and ascorbic acid. VN increased the activities of antioxidant enzymes in normal embryonic liver cells (WRL68) including, superoxide dismutase (SOD) and glutathione peroxidase (GPX) compared with to H2O2 group. The ethanolic extract of VN showed cytotoxicity to HepG2 cells in a dose and time-dependent manner with IC50 66.46 μg/ml, 57.36 μg/ml and 65.12 μg/ml at 24, 48, and 72-hours incubation respectively, with no sensitivity in WRL68 cells. This was associated with significant elevation in lactate dehydrogenase (LDH) release in HepG2 cells. In addition, the activation of caspase-3 enzyme suggesting that the observed cytotoxicity was mediated via an intrinsic apoptosis pathway.
  
  
  CONCLUSIONS
  PASS-predicted plant activity could efficiently help in selecting a promising pharmaceutical leads with high accuracy and required antioxidant and antiproliferative properties. This is the first report on PASS-predicted VN activity.
authors.affiliation: Department of Anatomy, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. normadiah_mk@um.edu.my.
authors.forename: Farkaad A
authors.initials: FA
authors.lastname: Kadir
authors.forename: Normadiah M
authors.initials: NM
authors.lastname: Kassim
authors.forename: Mahmood A
authors.initials: MA
authors.lastname: Abdulla
authors.forename: Wageeh A
authors.initials: WA
authors.lastname: Yehye
mesh_terms: D000704:Analysis of Variance; D000972:Antineoplastic Agents, Phytogenic / Q000737:chemistry / Q000494:pharmacology*; D000975:Antioxidants / Q000737:chemistry / Q000494:pharmacology*; D053148:Caspase 3 / Q000378:metabolism; D048430:Cell Shape; D002470:Cell Survival / Q000187:drug effects; D019295:Computational Biology; D056945:Hep G2 Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D020128:Inhibitory Concentration 50; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D008315:Malondialdehyde / Q000378:metabolism; D010936:Plant Extracts / Q000737:chemistry* / Q000494:pharmacology*; D012984:Software; D020950:Vitex / Q000737:chemistry*
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D000975:Antioxidants; D010936:Plant Extracts; D008315:Malondialdehyde; D006861:Hydrogen Peroxide; D053148:Caspase 3