Cigarette Smoke Impairs A2AAdenosine Receptor Mediated Wound Repair through Up-regulation of Duox-1 Expression.
Cigarette smoke (CS) exposure and intrinsic factors such as the NADPH oxidases produce high levels of reactive oxygen species (ROS), ensuing inflammatory tissue injury. We previously demonstrated that CS-generated ROS, particularly hydrogen peroxide (H2O2), impaired adenosine stimulated wound repair. We hypothesized that CS exposure modulates expression of Dual oxidase 1 (Duox-1), a NADPH oxidases known to generate H2O2. To test this hypothesis, we used human bronchial epithelial cell line Nuli-1 and C57BL/6 mice. Cells were treated with 5% CS extract (CSE) for various periods of time, and mice were exposed to whole body CS for six weeks. Both CSE and CS treatment induced increased expression of Duox-1, and silencing of Doux-1 improved the rate of cell wound repair induced by CSE treatment. Nuli-1 cells pretreated with thapsigargin but not calcium ionophore exhibited increased Duox-1 mRNA expression. CSE treatment stimulated PKCα activation, which was effectively blocked by pretreatment with diphenylene iodonium, a NADPH oxidase inhibitor. Compared to control, lungs from CS-exposed mice showed a significant increase in PKCα activity and Duox-1 expression. Collectively, the data demonstrated that CS exposure upregulates expression of Duox-1 protein. This further leads to H2O2production and PKCα activation, inhibiting A2AAR-stimulated wound repair.
Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa FL, USA.
Zhi
Z
Tian
Division of Pulmonary, Critical Care, Sleep and Allergy, Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE, USA.
Hui
H
Zhang
Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa FL, USA.
Jendayi
J
Dixon
Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa FL, USA.
Nicole
N
Traphagen
Division of Pulmonary, Critical Care, Sleep and Allergy, Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE, USA.|Department of Environmental, Agricultural, and Occupational Health, College of Public Health, University of Nebraska Medical Center, Omaha, NE, USA.|Research Service, Omaha-Western Iowa Veterans Affairs Medical Center, Omaha, NE, USA.
Todd A
TA
Wyatt
Research Service, Omaha-Western Iowa Veterans Affairs Medical Center, Omaha, NE, USA.|Division of Gastroenterology and Hepatology, Department of Internal Medicine, University of Nebraska Medical Center, NE, USA.
Kusum
K
Kharbanda
Division of Pulmonary, Critical Care, Sleep and Allergy, Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE, USA.
Samantha
S
Simet Chadwick
Division of Allergy and Immunology, Department of Internal Medicine, College of Medicine, University of South Florida, Tampa FL, USA.
Narasaiah
N
Kolliputi
Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa FL, USA.|Division of Pulmonary, Critical Care, Sleep and Allergy, Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE, USA.|Division of Allergy and Immunology, Department of Internal Medicine, College of Medicine, University of South Florida, Tampa FL, USA.
Diane S
DS
Allen-Gipson
D000818:Animals; D001980:Bronchi / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D061207:Calcium Ionophores / Q000494:pharmacology; D002460:Cell Line; D045424:Complex Mixtures / Q000494:pharmacology*; D000074623:Dual Oxidases / Q000235:genetics* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009861:Onium Compounds / Q000494:pharmacology; D051571:Protein Kinase C-alpha / Q000235:genetics / Q000378:metabolism; D043705:Receptor, Adenosine A2A / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D012907:Smoking / Q000503:physiopathology; D019284:Thapsigargin / Q000494:pharmacology; D014026:Nicotiana / Q000737:chemistry* / Q000633:toxicity; D014945:Wound Healing / Q000187:drug effects
D061207:Calcium Ionophores; D045424:Complex Mixtures; D004791:Enzyme Inhibitors; D009861:Onium Compounds; D043705:Receptor, Adenosine A2A; D019284:Thapsigargin; C007517:diphenyleneiodonium; D006861:Hydrogen Peroxide; D000074623:Dual Oxidases; C519130:Duox1 protein, mouse; D051571:Protein Kinase C-alpha