title: 20-HETE-induced mitochondrial superoxide production and inflammatory phenotype in vascular smooth muscle is prevented by glucose-6-phosphate dehydrogenase inhibition.
abstract: 20-Hydroxyeicosatetraeonic acid (20-HETE) produced by cytochrome P-450 monooxygenases in NADPH-dependent manner is proinflammatory, and it contributes to the pathogenesis of systemic and pulmonary hypertension. In this study, we tested the hypothesis that inhibition of glucose-6-phosphate dehydrogenase (G6PD), a major source of NADPH in the cell, prevents 20-HETE synthesis and 20-HETE-induced proinflammatory signaling that promotes secretory phenotype of vascular smooth muscle cells. Lipidomic analysis indicated that G6PD inhibition and knockdown decreased 20-HETE levels in pulmonary arteries as well as 20-HETE-induced 1) mitochondrial superoxide production, 2) activation of mitogen-activated protein kinase 1 and 3, 3) phosphorylation of ETS domain-containing protein Elk-1 that activate transcription of tumor necrosis factor-α gene (Tnfa), and 4) expression of tumor necrosis factor-α (TNF-α). Moreover, inhibition of G6PD increased protein kinase G1α activity, which, at least partially, mitigated superoxide production and Elk-1 and TNF-α expression. Additionally, we report here for the first time that 20-HETE repressed miR-143, which suppresses Elk-1 expression, and miR-133a, which is known to suppress synthetic/secretory phenotype of vascular smooth muscle cells. In summary, our findings indicate that 20-HETE elicited mitochondrial superoxide production and promoted secretory phenotype of vascular smooth muscle cells by activating MAPK1-Elk-1, all of which are blocked by inhibition of G6PD.
authors.affiliation: Department of Pharmacology, New York Medical College School of Medicine, Valhalla, New York;
authors.forename: Anand
authors.initials: A
authors.lastname: Lakhkar
authors.affiliation: Department of Pharmacology, New York Medical College School of Medicine, Valhalla, New York;
authors.forename: Vidhi
authors.initials: V
authors.lastname: Dhagia
authors.affiliation: Department of Pharmacology, New York Medical College School of Medicine, Valhalla, New York;
authors.forename: Sachindra Raj
authors.initials: SR
authors.lastname: Joshi
authors.affiliation: Department of Pharmacology, New York Medical College School of Medicine, Valhalla, New York;
authors.forename: Katherine
authors.initials: K
authors.lastname: Gotlinger
authors.affiliation: Department of Physiology, New York Medical College School of Medicine, Valhalla, New York; and.
authors.forename: Dhara
authors.initials: D
authors.lastname: Patel
authors.affiliation: Department of Physiology, New York Medical College School of Medicine, Valhalla, New York; and.
authors.forename: Dong
authors.initials: D
authors.lastname: Sun
authors.affiliation: Department of Physiology, New York Medical College School of Medicine, Valhalla, New York; and Translational Centre for Pulmonary Hypertension, New York Medical College School of Medicine, Valhalla, New York.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Wolin
authors.affiliation: Department of Pharmacology, New York Medical College School of Medicine, Valhalla, New York;
authors.forename: Michal L
authors.initials: ML
authors.lastname: Schwartzman
authors.affiliation: Department of Pharmacology, New York Medical College School of Medicine, Valhalla, New York; Translational Centre for Pulmonary Hypertension, New York Medical College School of Medicine, Valhalla, New York s_gupte@nymc.edu.
authors.forename: Sachin A
authors.initials: SA
authors.lastname: Gupte
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D001013:Aorta, Thoracic / Q000187:drug effects / Q000201:enzymology; D002417:Cattle; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism; D042926:Cytochrome P-450 CYP4A / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005838:Genotype; D005954:Glucosephosphate Dehydrogenase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D006893:Hydroxyeicosatetraenoic Acids / Q000378:metabolism*; D007249:Inflammation / Q000201:enzymology / Q000235:genetics / Q000517:prevention & control*; D018836:Inflammation Mediators / Q000378:metabolism; D008297:Male; D008817:Mice, Mutant Strains; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D008931:Mitochondria, Muscle / Q000187:drug effects* / Q000201:enzymology; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000201:enzymology; D009154:Mutation; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000201:enzymology; D010641:Phenotype; D011651:Pulmonary Artery / Q000187:drug effects / Q000201:enzymology; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000187:drug effects; D013481:Superoxides / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism; D050807:ets-Domain Protein Elk-1 / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; C494114:Elk1 protein, rat; D004791:Enzyme Inhibitors; D006893:Hydroxyeicosatetraenoic Acids; D018836:Inflammation Mediators; C569508:MIRN143 microRNA, rat; D035683:MicroRNAs; D014409:Tumor Necrosis Factor-alpha; D050807:ets-Domain Protein Elk-1; D013481:Superoxides; C055987:20-hydroxy-5,8,11,14-eicosatetraenoic acid; D005954:Glucosephosphate Dehydrogenase; D042926:Cytochrome P-450 CYP4A; D017869:Cyclic GMP-Dependent Protein Kinases; D048049:Extracellular Signal-Regulated MAP Kinases
keywords: 20-HETE; TNF-α; elk-1; extracellular signal regulated kinase 2 and 1; miRs; mitogen-activated protein kinase 1 and 3; protein kinase G