Cytosolic calcium increase in coronary endothelial cells after H2O2 exposure and the inhibitory effect of U78517F.
1. Cytosolic calcium concentrations ([Ca2+]i) were determined with fura-2 on both resting (unstimulated) and A23187-stimulated coronary endothelial cells following injury by hydrogen peroxide (H2O2). 2. Treatment of cells with H2O2 (10(-4) M) caused an increase in the resting [Ca2+]i, which reached a maximum of five fold after 3 h. 3. The increase in resting [Ca2+]i was significantly attenuated by treatment with U78517F, a potent inhibitor of lipid peroxidation, at a concentration of 10(-6) M or greater. Catalase (50 u ml-1) also markedly inhibited the H2O2-induced rise in [Ca2+]i. Pretreatment with verapamil (10(-5) M), nifedipine (10(-6) M) or diltiazem (10(-5) M) had no effect on the increase in [Ca2+]i following addition of H2O2. 4. A23187 produced a transient increase in [Ca2+]i followed by a sustained plateau. The initial peak and plateau phase responses to A23187 were augmented by H2O2. This augmentation of [Ca2+]i was suppressed by U78517F or catalase but not by Ca-entry blockers. 5. Thus, it is likely that lipid peroxidation plays a critical role in the sustained increase in [Ca2+]i that occurs following treatment with H2O2 and that this continues in the presence of agonists which stimulate the endothelium. Voltage-gated Ca2+ channels do not seem to be involved in the genesis of cellular damage associated with sustained large increases in [Ca2+]i.
Kimura
M
M
Department of Pharmacology, Tsukuba Research Laboratories, Upjohn Pharmaceuticals Limited, Japan.
Maeda
K
K
Hayashi
S
S
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002374:Catalase / Q000494:pharmacology; D002478:Cells, Cultured; D002839:Chromans / Q000494:pharmacology*; D003331:Coronary Vessels / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D004110:Diltiazem / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D016166:Free Radical Scavengers; D006861:Hydrogen Peroxide / Q000633:toxicity*; D066298:In Vitro Techniques; D015227:Lipid Peroxidation / Q000187:drug effects*; D009543:Nifedipine / Q000494:pharmacology; D010879:Piperazines / Q000494:pharmacology*; D013552:Swine; D014700:Verapamil / Q000494:pharmacology
D002839:Chromans; D016166:Free Radical Scavengers; D010879:Piperazines; C066382:U 78517F; D006861:Hydrogen Peroxide; D014700:Verapamil; D002374:Catalase; D004110:Diltiazem; D009543:Nifedipine; D002118:Calcium