title: Preconditioning by isoflurane elicits mitochondrial protective mechanisms independent of sarcolemmal KATP channel in mouse cardiomyocytes.
abstract: Cardiac mitochondria and the sarcolemmal (sarc)KATP channels contribute to cardioprotective signaling of anesthetic-induced preconditioning. Changes in mitochondrial bioenergetics influence the sarcolemmal ATP-sensitive K (sarcKATP) channel function, but whether this channel has impacts on mitochondria is uncertain. We used the mouse model with deleted pore-forming Kir6.2 subunit of sarcKATP channel (Kir6.2 KO) to investigate whether the functional sarcKATP channels are necessary for isoflurane activation of mitochondrial protective mechanisms. Ventricular cardiomyocytes were isolated from C57Bl6 wild-type (WT) and Kir6.2 KO mouse hearts. Flavoprotein autofluorescence, mitochondrial reactive oxygen species production, and mitochondrial membrane potential were monitored by laser-scanning confocal microscopy in intact cardiomyocytes. Cell survival was assessed using H2O2-induced stress. Isoflurane (0.5 mM) increased flavoprotein fluorescence to 180% ± 14% and 190% ± 15% and reactive oxygen species production to 118% ± 2% and 124% ± 6% of baseline in WT and Kir6.2 KO myocytes, respectively. Tetramethylrhodamine ethyl ester fluorescence decreased to 84% ± 6% in WT and to 86% ± 4% in Kir6.2 KO myocytes. This effect was abolished by 5HD. Pretreatment with isoflurane decreased the stress-induced cell death from 31% ± 1% to 21% ± 1% in WT and from 44% ± 2% to 35% ± 2% in Kir6.2 KO myocytes. In conclusion, Kir6.2 deletion increases the sensitivity of intact cardiomyocytes to oxidative stress, but does not alter the isoflurane-elicited protective mitochondrial mechanisms, suggesting independent roles for cardiac mitochondria and sarcKATP channels in anesthetic-induced preconditioning by isoflurane.
authors: Department of Anesthesiology, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
authors: Maria
authors: M
authors: Muravyeva
authors: Filip
authors: F
authors: Sedlic
authors: Nicholas
authors: N
authors: Dolan
authors: Zeljko J
authors: ZJ
authors: Bosnjak
authors: Anna
authors: A
authors: Stadnicka
mesh_terms: D018685:Anesthetics, Inhalation / Q000494:pharmacology*; D000818:Animals; D002470:Cell Survival; D005420:Flavoproteins / Q000187:drug effects / Q000502:physiology; D005453:Fluorescence; D006352:Heart Ventricles / Q000166:cytology; D019157:Ischemic Preconditioning, Myocardial; D007530:Isoflurane / Q000494:pharmacology*; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D008929:Mitochondria, Heart / Q000187:drug effects* / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000187:drug effects*; D018384:Oxidative Stress / Q000187:drug effects; D024661:Potassium Channels, Inwardly Rectifying / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D012508:Sarcolemma / Q000187:drug effects*; D000099161:Kcnj11 Channel
chemical_list: D018685:Anesthetics, Inhalation; D005420:Flavoproteins; D007530:Isoflurane; D024661:Potassium Channels, Inwardly Rectifying; D017382:Reactive Oxygen Species; D000099161:Kcnj11 Channel