Intact smooth muscle metabolism: its responses to cyanide poisoning and pyruvate stimulation.
Smooth muscle mitochondria are unique in their compartmentalization of metabolism with the contractile proteins and putative role in cell fate choice. In this study we examine the relative and quantitative differences that smooth muscle mitochondria have with regard to cyanide inhibition. The effect of cyanide poisoning in the mitochondria of smooth muscle was examined in the intact porcine carotid artery and intact guinea pig stomach. The respiratory responses of these tissues were monitored in the presence of cyanide and following the addition of various metabolites. In HEPES buffer with 10 mM glucose as the substrate, it was found that the EC50 for cyanide inhibition was 0.11+/-0.02 and 0.14+/-0.02 mM in the pig carotid and guinea stomach respectively. We also found that the signaling metabolite, pyruvate could partially reverse this inhibition. With pyruvate (10 mM) as the substrate, the EC50 increased significantly to 6.52+/-0.11 (carotid artery) and 1.95+/-0. 30 (stomach) mM as well as being significantly different between the tissues. This apparent resistance to cyanide inhibition caused by pyruvate was lost when sodium bicarbonate buffer was used (EC50 in the presence of pyruvate of 0.13+/-0.04 and 0.25+/-0.04 for carotid and stomach respectively). HEPES buffer permitted pyruvate's protection from cyanide poisoning whereas bicarbonate buffer did not. The rate of respiration caused by pyruvate re-stimulation was not significantly different than control in the stomach (2.63+/-0.77 and 2.69+/-0.3??Mol O2/min/g dw respectively) but was significantly greater in the carotid artery. Therefore, smooth muscle with 10 mM pyruvate and 1 mM cyanide had a rate or respiration significantly greater than with 10 mM glucose alone (0.90+/-0.2 and 0.59+/-0. 06??Mol O2/min/g dw respectively). Using 31P NMR spectroscopy, we observed a complete normalisation of high energy phosphates and pH in the guinea pig stomach smooth muscle caused by pyruvate after cyanide poisoning. These results suggest that cyanide's toxicity of smooth muscle oxidative metabolism is affected by the buffer (HEPES versus Bicarbonate) and metabolic signalling molecules or substrates (pyruvate versus glucose) in which the tissues are exposed to as well as tissue to tissue variations.
Clark
J F
JF
Department of Biochemistry, University of Oxford, OX1 3QU England. joseph.clark@uc.edu
Matsumoto
T
T
Nakayama
S
S
D000818:Animals; D002021:Buffers; D002339:Carotid Arteries; D019069:Cell Respiration / Q000187:drug effects; D003486:Cyanides / Q000037:antagonists & inhibitors / Q000506:poisoning*; D003576:Electron Transport Complex IV / Q000037:antagonists & inhibitors / Q000378:metabolism; D006168:Guinea Pigs; D066298:In Vitro Techniques; D007928:Lethal Dose 50; D009682:Magnetic Resonance Spectroscopy; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology*; D009130:Muscle, Smooth / Q000187:drug effects / Q000378:metabolism*; D010101:Oxygen Consumption / Q000187:drug effects; D011773:Pyruvates / Q000494:pharmacology; D013270:Stomach; D013552:Swine
D002021:Buffers; D003486:Cyanides; D011773:Pyruvates; D003576:Electron Transport Complex IV