ET-1 stimulates pulmonary arterial smooth muscle cell proliferation via induction of reactive oxygen species.
Recent studies implicate reactive oxygen species (ROS) such as superoxide anions and H(2)O(2) in the proliferation of systemic vascular smooth muscle cells (SMCs). However, the role of ROS in SMC proliferation within the pulmonary circulation remains unclear. We investigated the effects of endothelin-1 (ET-1), a potential SMC mitogen, on ROS production and proliferation of fetal pulmonary artery SMCs (FPASMCs). Exposure to ET-1 resulted in increases in superoxide production and viable FPASMCs after 72 h. These increases were prevented by pretreatment with PD-156707. Treatment with pertussis toxin blocked the effects of ET-1, whereas cholera toxin stimulated superoxide production and increased viable cell numbers even in the absence of ET-1. Wortmannin, LY-294002, diphenyleneiodonium (DPI), 4-(2-aminoethyl)benzenesulfonyl fluoride, and apocynin also prevented the ET-1-mediated increases in superoxide production and viable cell numbers. Exposure to H(2)O(2) or diethyldithiocarbamate increased viable cell number by 37% and 50%, respectively. Conversely, ascorbic acid and DPI decreased viable cell number, which appeared to be due to an increase in programmed cell death. Our data suggest that ET-1 exerts a mitogenic effect on FPASMCs via an increase in ROS production and that antioxidants can block this effect via induction of apoptosis. Antioxidant treatment may therefore represent a potential therapy for pulmonary vascular diseases.
Wedgwood
S
S
Department of Pediatrics, Northwestern University Medical School, Chicago, Illinois 60611-3008, USA.
Dettman
R W
RW
Black
S M
SM
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D001205:Ascorbic Acid / Q000494:pharmacology; D002455:Cell Division / Q000502:physiology*; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D016895:Culture Media, Serum-Free; D004149:Dioxoles / Q000494:pharmacology; D004050:Ditiocarb / Q000494:pharmacology; D019332:Endothelin-1 / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004996:Ethidium / Q000031:analogs & derivatives* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects* / Q000196:embryology; D011651:Pulmonary Artery / Q000166:cytology / Q000187:drug effects* / Q000196:embryology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012756:Sheep / Q000196:embryology; D015398:Signal Transduction / Q000502:physiology
D000975:Antioxidants; D002614:Chelating Agents; D016895:Culture Media, Serum-Free; D004149:Dioxoles; D019332:Endothelin-1; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; C093321:PD 156707; D017382:Reactive Oxygen Species; C067883:dihydroethidium; D004050:Ditiocarb; D004996:Ethidium; D001205:Ascorbic Acid