title: Free radicals mediate actions of oxyhemoglobin on cerebrovascular smooth muscle cells.
abstract: Single smooth muscle cells were isolated from the basilar artery of the rat by enzymatic dispersion. The membrane properties of the cells were assessed using the patch-electrode voltage-clamp technique, and cell viability was monitored using fluorescein diacetate uptake. Exposure of the cells to oxyhemoglobin (5 microM) resulted in 1) contraction, 2) the appearance of membrane blebs, 3) an increase in the outward potassium currents, 4) a decrease in the membrane resistance, and 5) cell death. In contrast, no effect of oxyhemoglobin on cultured murine neuroblastoma cells was observed. Methemoglobin (100 microM) had no effects on the smooth muscle cells. Catalase (300 units/ml) or dimethyl sulfoxide (0.5%) protected against the effects of oxyhemoglobin; superoxide dismutase (100-1,000 units/ml) provided only partial protection. Exposure of the cells to superoxide anions generated by xanthine (1 mM) plus xanthine oxidase (10 units/l) or to hydrogen peroxide (500 microM) caused an increase in the outward potassium currents without affecting membrane resistance. Generation of hydroxyl radicals by metal ions plus hydrogen peroxide caused the same effects as oxyhemoglobin, that is, an increase in the potassium currents, followed by a decrease in the membrane resistance and cell death. In conclusion, it appears that oxyhemoglobin exerts its effects on vascular smooth muscle cells by the generation of free radicals, chiefly hydroxyl radicals.
authors.affiliation: Department of Surgery, University of Alberta, Edmonton, Canada.
authors.forename: J A
authors.initials: JA
authors.lastname: Steele
authors.forename: N
authors.initials: N
authors.lastname: Stockbridge
authors.forename: G
authors.initials: G
authors.lastname: Maljkovic
authors.forename: B
authors.initials: B
authors.lastname: Weir
mesh_terms: D000818:Animals; D002374:Catalase / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D002560:Cerebrovascular Circulation / Q000187:drug effects*; D004594:Electrophysiology; D005260:Female; D005609:Free Radicals; D051379:Mice; D009119:Muscle Contraction; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D010100:Oxygen / Q000502:physiology*; D010108:Oxyhemoglobins / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D011188:Potassium / Q000502:physiology; D051381:Rats; D011919:Rats, Inbred Strains; D013482:Superoxide Dismutase / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects
chemical_list: D005609:Free Radicals; D010108:Oxyhemoglobins; D002374:Catalase; D013482:Superoxide Dismutase; D011188:Potassium; D010100:Oxygen