Isoflurane preserves adenosine triphosphate levels in anoxic isolated rat hepatocytes by stimulating glycolytic adenosine triphosphate formation.
The hypothesis that general anesthetics protect energy reserves by decreasing energy demand is widely accepted but poorly substantiated. Isoflurane at clinical doses preserved adenosine triphosphate (ATP) levels in anoxic isolated hepatocytes. Specific inhibitors were used to block mitochondrial and/or glycolytic ATP formation to ascertain whether pathways of energy supply or demand, or both, were involved in ATP preservation by isoflurane. Hepatocytes were isolated from fed adult male rats after perfusing livers with Krebs buffer containing collagenase. Cells were incubated in Krebs buffer for 0-30 min at 25 degrees C under N2/CO2 (95%/5%) +/- isoflurane 0.63 mM in liquid phase. Oligomycin, iodoacetate, or fasting were used to block mitochondrial and glycolytic ATP formation. Under anoxia alone, ATP levels declined more slowly in the presence than in the absence of isoflurane, confirming the ATP-protective effect of isoflurane reported previously. With oligomycin plus iodoacetate blocking all ATP formation, ATP decline (representing pure ATP consumption) was not slowed by isoflurane. Isoflurane's protective effect recurred when glycolytic ATP supply was restored by incubating with oligomycin only. The protective effect was accompanied by increased lactate accumulation, and both effects-ATP preservation and lactate formation-were similarly dependent on isoflurane concentration. We conclude that the protective effect of isoflurane on energy status in anoxic isolated hepatocytes was not associated with reduced ATP demand but with enhanced ATP supply via stimulation of glycolysis.
Matsushita
M
M
Department of Anesthesiology, University of Nebraska Medical Center, Omaha 68198-4455, USA.
Ohashi
I
I
Becker
G L
GL
Pohorecki
R
R
D000255:Adenosine Triphosphate / Q000096:biosynthesis / Q000378:metabolism*; D018685:Anesthetics, Inhalation / Q000494:pharmacology*; D000818:Animals; D015687:Cell Hypoxia / Q000502:physiology; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006019:Glycolysis / Q000187:drug effects*; D007461:Iodoacetates / Q000494:pharmacology; D019807:Iodoacetic Acid; D007530:Isoflurane / Q000494:pharmacology*; D008099:Liver / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D009840:Oligomycins / Q000494:pharmacology; D051381:Rats; D012966:Sodium Cyanide / Q000494:pharmacology; D013268:Stimulation, Chemical; D014475:Uncoupling Agents / Q000494:pharmacology
D018685:Anesthetics, Inhalation; D004791:Enzyme Inhibitors; D007461:Iodoacetates; D009840:Oligomycins; D014475:Uncoupling Agents; D000255:Adenosine Triphosphate; D007530:Isoflurane; D012966:Sodium Cyanide; D019807:Iodoacetic Acid