Isoflurane protects against human endothelial cell apoptosis by inducing sphingosine kinase-1 via ERK MAPK.
Endothelial dysfunction is a major clinical problem affecting virtually every patient requiring critical care. Volatile anesthetics are frequently used during the perioperative period and protect the heart and kidney against ischemia and reperfusion injury. We aimed to determine whether isoflurane, the most commonly used volatile anesthetic in the USA, protects against endothelial apoptosis and necrosis and the mechanisms involved in this protection. Human endothelial EA.hy926 cells were pretreated with isoflurane or carrier gas (95% room air + 5% CO(2)) then subjected to apoptosis with tumor necrosis factor-α or to necrosis with hydrogen peroxide. DNA laddering and in situ Terminal Deoxynucleotidyl Transferase Biotin-dUTP Nick-End Labeling (TUNEL) staining determined EA.hy926 cell apoptosis and percent LDH released determined necrosis. We also determined whether isoflurane modulates the expression and activity of sphingosine kinase-1 (SK1) and induces the phosphorylation of extracellular signal regulated kinase (ERK MAPK) as both enzymes are known to protect against cell death. Isoflurane pretreatment significantly decreased apoptosis in EA.hy926 cells as evidenced by reduced TUNEL staining and DNA laddering without affecting necrosis. Mechanistically, isoflurane induces the phosphorylation of ERK MAPK and increased SK1 expression and activity in EA.hy926 cells. Finally, selective blockade of SK1 (with SKI-II) or S1P(1) receptor (with W146) abolished the anti-apoptotic effects of isoflurane. Taken together, we demonstrate that isoflurane, in addition to its potent analgesic and anesthetic properties, protects against endothelial apoptosis most likely via SK1 and ERK MAPK activation. Our findings have significant clinical implication for protection of endothelial cells during the perioperative period and patients requiring critical care.
Department of Anesthesiology, College of Physicians and Surgeons of Columbia University, P&S Box 46 (PH-5), 630 West 168th Street, New York, NY 10032, USA.|Department of Pediatrics, College of Physicians and Surgeons of Columbia University, CHN2-255, 3959 Broadway, New York, NY 10032, USA.
Adnan M
AM
Bakar
Department of Anesthesiology, College of Physicians and Surgeons of Columbia University, P&S Box 46 (PH-5), 630 West 168th Street, New York, NY 10032, USA.
Sang Won
SW
Park
Department of Anesthesiology, College of Physicians and Surgeons of Columbia University, P&S Box 46 (PH-5), 630 West 168th Street, New York, NY 10032, USA.
Mihwa
M
Kim
Department of Anesthesiology, College of Physicians and Surgeons of Columbia University, P&S Box 46 (PH-5), 630 West 168th Street, New York, NY 10032, USA.
H Thomas
HT
Lee
D018685:Anesthetics, Inhalation / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D053938:DNA Fragmentation / Q000187:drug effects; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005419:Flavonoids / Q000494:pharmacology; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D007530:Isoflurane / Q000494:pharmacology*; D009336:Necrosis / Q000209:etiology; D010766:Phosphorylation / Q000187:drug effects; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D000099157:Sphingosine Kinase
D018685:Anesthetics, Inhalation; D048049:Extracellular Signal-Regulated MAP Kinases; D005419:Flavonoids; D006861:Hydrogen Peroxide; D007530:Isoflurane; D017853:Phosphotransferases (Alcohol Group Acceptor); D014409:Tumor Necrosis Factor-alpha; D000099157:Sphingosine Kinase; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one