title: Evaluation of Protein Ubiquitylation in Heart Tissue of Rats Exposed to Diazinon (an Organophosphate Insecticide) and Crocin (an Active Saffron Ingredient): Role of HIF-1α.
abstract: BACKGROUND
  Organophosphate compounds, such as diazinon (DZN), are widely used in agriculture and can lead to formation of reactive oxygen species (ROS) in cardiovascular system. ROS are highly toxic since they can cause serious changes in proteins including ubiquitylation. Crocin (a carotenoid isolated from saffron), has protective effects against DZN cardiotoxicity. In this study level of total protein ubiquitylation as markers of oxidative stress and level of ubiquitin-HIF-1α and P53, known substrates of ubiquitylation, in rat hearts exposed to DZN and crocin were evaluated.
  
  
  METHODS
  Rats were divided into 7 groups: corn oil (control), DZN (15 mg/kg/day, gavage), crocin (12.5, 25, 50 mg/kg/day, i. p.) plus DZN, vitamin E (200 IU/kg, i. p., 3 days a week) plus DZN and crocin (50 mg/kg/day, i. p.). Treatments were continued for 4 weeks. Total protein ubiquitylation, total HIF-1α and P53 were analyzed by western blotting. Total HIF-1α and P53 were purified by immunoprecipitation (IP) and ubiquitin- HIF-1α and P53 were analyzed by western blotting.
  
  
  RESULTS
  Higher protein ubiquitylation levels were observed in DZN treated rats. Decrease in ubiquitin-HIF-1α was also shown, and leads to higher HIF-1α protein levels in DZN group. Crocin (50 mg/kg) and vit. E protected cells against DZN protein ubiquitylation. Significant differences were not observed between the ubiquitin - P53 and total P53 protein levels.
  
  
  CONCLUSIONS
  Our results showed that ubiquitylation could be considered as a marker of oxidative stress in rats exposed to DZN. Increase in level of HIF-α may compensate adverse effect of DNZ in rat heart.
authors.affiliation: Targeted Drug Delivery Research Center, Department of Pharmacodinamy and Toxicology, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: B M
authors.initials: BM
authors.lastname: Razavi
authors.affiliation: Pharmaceutical Research Center, Department of Pharmacodinamy and Toxicology, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: H
authors.initials: H
authors.lastname: Hosseinzadeh
authors.affiliation: Targeted Drug Delivery Research Center, Department of Pharmacodinamy and Toxicology, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: M
authors.initials: M
authors.lastname: Imenshahidi
authors.affiliation: Targeted Drug Delivery Research Center, Department of Pharmacodinamy and Toxicology, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: M
authors.initials: M
authors.lastname: Malekian
authors.affiliation: Nanotechnology Research Center, Department of Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: M
authors.initials: M
authors.lastname: Ramezani
authors.affiliation: Pharmaceutical Research Center, Department of Medicinal Chemistry, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: K
authors.initials: K
authors.lastname: Abnous
mesh_terms: D000818:Animals; D002316:Cardiotonic Agents / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology; D066126:Cardiotoxicity / Q000517:prevention & control*; D002338:Carotenoids / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*; D027622:Crocus / Q000737:chemistry; D003976:Diazinon / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D007306:Insecticides / Q000633:toxicity; D008297:Male; D018384:Oxidative Stress / Q000187:drug effects*; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D054875:Ubiquitination / Q000187:drug effects; D014810:Vitamin E / Q000494:pharmacology
chemical_list: D002316:Cardiotonic Agents; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D007306:Insecticides; D017382:Reactive Oxygen Species; D016159:Tumor Suppressor Protein p53; D014810:Vitamin E; D002338:Carotenoids; C029036:crocin; D003976:Diazinon