title: Intracellular mechanisms of constriction of rat aorta by ethanol.
abstract: The intracellular mechanisms mediating vasoconstriction by ethanol are poorly understood. This investigation was designed to provide evidence on the role of protein kinase C (PKC) and calmodulin in vasoconstriction by ethanol. We studied helically cut strips of rat aorta that were exposed to ethanol before and in the presence of the PKC inhibitors calphostin C (79, 239, and 798 nM) or 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H7, 10 microM), and the calmodulin inhibitor, trifluoperazine (TFP, 10 microM). To test for the specificity of the PKC inhibitors, we measured the responses of aortas to potassium and phorbol 12-myristate 13-acetate (PMA) in the absence and presence of calphostin C and H7. To test for the specificity of TFP, we measured the responses of aortas to serotonin, potassium, PMA, and the thromboxane A2 mimic. 9,11-dideoxy-11 alpha, 9 alpha-epoxy-methanoprostaglandin F2a (U46619), in the absence and presence of TFP. We also studied the effect of the combination of calphostin C and TFP on constriction of the aorta by ethanol. We also measured the importance of intracellular and extracellular calcium in constriction of the aorta by ethanol. Force generation was measured before, and then during exposure of the strips to calcium-free buffer with EGTA, or calcium-free buffer with EGTA plus caffeine. We found that both PKC inhibitors antagonized vasoconstriction by ethanol and PMA. However, H7 antagonized contractions by potassium, but calphostin C did not. We found that TFP caused 99 +/- 1% inhibition of maximum contraction to serotonin, 90 +/- 4% inhibition of maximum contraction to potassium, 63 +/- 6% inhibition of maximum contraction to PMA, and 8 +/- 5% inhibition of maximum contraction to U46619. TFP caused a 22 +/- 8% inhibition of contraction to ethanol. The combination of TFP and calphostin C antagonized vasoconstriction by ethanol to a degree similar to that of calphostin C alone. We also found that contractions to ethanol were only 16 +/- 7% of control values in a calcium-free plus EGTA buffer. Contractions to ethanol were 0 +/- 1% of control values in calcium-free buffer with EGTA plus caffeine. We conclude that: 1-vasoconstriction by ethanol is, at least in part, mediated by PKC; 2-constriction by ethanol is mediated to a minimal extent by calmodulin, and 3-part of the constriction by ethanol of the aorta is mediated by a caffeine-sensitive pool of intracellular calcium.
authors.affiliation: Department of Pharmacology and Neuroscience A136, Albany Medical College, NY 12208, USA. andrew_werber@ccgateway.amc.edu
authors.forename: A H
authors.initials: AH
authors.lastname: Werber
authors.forename: R A
authors.initials: RA
authors.lastname: Morgan
authors.forename: P
authors.initials: P
authors.lastname: Zhou
authors.forename: C
authors.initials: C
authors.lastname: Yang
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000494:pharmacology; D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000502:physiology; D002147:Calmodulin / Q000037:antagonists & inhibitors / Q000502:physiology; D002492:Central Nervous System Depressants / Q000494:pharmacology*; D000697:Central Nervous System Stimulants / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D000431:Ethanol / Q000494:pharmacology*; D066298:In Vitro Techniques; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000187:drug effects*; D009281:Naphthalenes / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014268:Trifluoperazine / Q000494:pharmacology; D014661:Vasoconstriction / Q000187:drug effects*
chemical_list: D002147:Calmodulin; D002492:Central Nervous System Depressants; D000697:Central Nervous System Stimulants; D004791:Enzyme Inhibitors; D009281:Naphthalenes; D014268:Trifluoperazine; D002110:Caffeine; D000431:Ethanol; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D011493:Protein Kinase C; C058819:calphostin C; D002118:Calcium