title: Dextran sulfate protects porcine but not bovine cultured endothelial cells from free radical injury.
abstract: Previous studies demonstrated that the polyanion dextran sulfate (DS) protects rat coronary and porcine aortic endothelium (PAE) from oxygen-derived free radical (OFR) injury due to hydrogen peroxide (H2O2) or xanthine/xanthine oxidase (X/XO). To determine if DS has a similar protective effect in bovine aortic endothelium (BAE) and bovine brain microvascular endothelium (BBME), H2O2 or X/XO was added to confluent cultures. Cell injury was assessed 1 d later by measuring the percentage of viable cells (by trypan blue exclusion) and the release of lactate dehydrogenase (LDH) into the medium. After H2O2 doses of 6.0 mM for BAE and BBME and 0.8 mM for PAE, and after X doses of 10 microM and XO doses of 0.3 U/mL for all cell types, approximately 50% of cells were viable. Cultures were pretreated with DS (0.001 to 500 microg/mL) 24 to 26 h prior to H2O2 or X/XO exposure. Pretreatment at concentrations of 0.5, 5, and 50 microg/mL significantly increased the percentage of viable cells and reduced LDH release in cultures of PAE, but not BAE or BBME, treated with H2O2. Similarly, pretreatment with DS concentrations of 5 and 50 microg/mL significantly increased the percentage of viable cells and reduced LDH release in cultures of PAE, but not BAE or BBME, treated with X/XO. Thus, DS protected porcine but not bovine endothelium. Catalase (10 U/mL) increased the percentage of viable cells and reduced LDH release in H2O2-treated BAE and BBME, suggesting that DS likely acts by a different mechanism and does not neutralize H2O2. These results suggest that the protective effect of DS on OFR-injured endothelium is species-dependent.
authors.affiliation: Toxicology Centre, Saskatoon, Saskatchewan.
authors.forename: Jason I
authors.initials: JI
authors.lastname: Ram
authors.forename: Linda M
authors.initials: LM
authors.lastname: Hiebert
mesh_terms: D000818:Animals; D002417:Cattle; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D016264:Dextran Sulfate / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects*; D005609:Free Radicals / Q000633:toxicity; D006861:Hydrogen Peroxide / Q000633:toxicity*; D013045:Species Specificity; D013552:Swine; D019820:Xanthine / Q000633:toxicity*; D014969:Xanthine Oxidase / Q000633:toxicity*
chemical_list: D005609:Free Radicals; D019820:Xanthine; D016264:Dextran Sulfate; D006861:Hydrogen Peroxide; D014969:Xanthine Oxidase