Hyperoxia-induced NAD(P)H oxidase activation and regulation by MAP kinases in human lung endothelial cells.
Hyperoxia increases reactive oxygen species (ROS) production in vascular endothelium; however, the mechanisms involved in ROS generation are not well characterized. We determined the role and regulation of NAD(P)H oxidase in hyperoxia-induced ROS formation in human pulmonary artery endothelial cells (HPAECs). Exposure of HPAECs to hyperoxia for 1, 3, and 12 h increased the generation of superoxide anion, which was blocked by diphenyleneiodonium but not by rotenone or oxypurinol. Furthermore, hyperoxia enhanced NADPH- and NADH-dependent and superoxide dismutase- or diphenyleneiodonium-inhibitable ROS production in HPAECs. Immunohistocytochemistry and Western blotting revealed the presence of gp91, p67 phox, p22 phox, and p47 phox subcomponents of NADPH oxidase in HPAECs. Transfection of HPAECs with p22 phox antisense plasmid inhibited hyperoxia-induced ROS production. Exposure of HPAECs to hyperoxia activated p38 MAPK and ERK, and inhibition of p38 MAPK and MEK1/2 attenuated the hyperoxia-induced ROS generation. These results suggest a role for MAPK in regulating hyperoxia-induced NAD(P)H oxidase activation in HPAECs.
Parinandi
Narasimham L
NL
Division of Pulmonary and Critical Care Medicine and Division of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21224, USA.
Kleinberg
Michael A
MA
Usatyuk
Peter V
PV
Cummings
Rhett J
RJ
Pennathur
Arjun
A
Cardounel
Arturo J
AJ
Zweier
Jay L
JL
Garcia
Joe G N
JG
Natarajan
Viswanathan
V
D016375:Antisense Elements (Genetics) / Q000494:pharmacology; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000201:enzymology*; D004789:Enzyme Activation / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D018496:Hyperoxia / Q000201:enzymology* / Q000503:physiopathology; D026901:Membrane Transport Proteins; D020928:Mitogen-Activated Protein Kinases / Q000502:physiology*; D009252:NADPH Dehydrogenase / Q000235:genetics; D019255:NADPH Oxidases / Q000378:metabolism* / Q000502:physiology; D009861:Onium Compounds / Q000494:pharmacology; D010587:Phagocytosis / Q000502:physiology; D010750:Phosphoproteins / Q000235:genetics; D011651:Pulmonary Artery / Q000201:enzymology; D017382:Reactive Oxygen Species / Q000037:antagonists & inhibitors / Q000378:metabolism
D016375:Antisense Elements (Genetics); D004791:Enzyme Inhibitors; D026901:Membrane Transport Proteins; D009861:Onium Compounds; D010750:Phosphoproteins; D017382:Reactive Oxygen Species; C007517:diphenyleneiodonium; D006861:Hydrogen Peroxide; D019255:NADPH Oxidases; C086986:CYBA protein, human; D009252:NADPH Dehydrogenase; D020928:Mitogen-Activated Protein Kinases