Attenuation of lipopolysaccharide-induced oxidative stress and apoptosis in fetal pulmonary artery endothelial cells by hypoxia.
Pulmonary vascular endothelial injury resulting from lipopolysaccharide (LPS) and oxygen toxicity contributes to vascular simplification seen in the lungs of premature infants with bronchopulmonary dysplasia. Whether the severity of endotoxin-induced endothelial injury is modulated by ambient oxygen tension (hypoxic intrauterine environment vs. hyperoxic postnatal environment) remains unknown. We posited that ovine fetal pulmonary artery endothelial cells (FPAEC) will be more resistant to LPS toxicity under hypoxic conditions (20-25 Torr) mimicking the fetal milieu. LPS (10 microg/ml) inhibited FPAEC proliferation and induced apoptosis under normoxic conditions (21% O(2)) in vitro. LPS-induced FPAEC apoptosis was attenuated in hypoxia (5% O(2)) and exacerbated by hyperoxia (55% O(2)). LPS increased intracellular superoxide formation, as measured by 2-hydroxyethidium (2-HE) formation, in FPAEC in normoxia and hypoxia. 2-HE formation in LPS-treated FPAEC increased in parallel with the severity of LPS-induced apoptosis in FPAEC, increasing from hypoxia to normoxia to hyperoxia. Differences in LPS-induced apoptosis between hypoxia and normoxia were abolished when LPS-treated FPAEC incubated in hypoxia were pretreated with menadione to increase superoxide production. Apocynin decreased 2-HE formation, and attenuated LPS-induced FPAEC apoptosis under normoxic conditions. We conclude that ambient oxygen concentration modulates the severity of LPS-mediated injury in FPAEC by regulating superoxide levels produced in response to LPS.
Sampath
Venkatesh
V
Department of Pediatrics, Medical College of Wisconsin, Milwaukee, USA. vsampath@mcw.edu
Radish
Aaron C
AC
Eis
Annie L
AL
Broniowska
Katarzyna
K
Hogg
Neil
N
Konduri
Girija G
GG
D000098:Acetophenones / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001997:Bronchopulmonary Dysplasia / Q000201:enzymology* / Q000473:pathology / Q000517:prevention & control; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D042783:Endothelial Cells / Q000473:pathology / Q000502:physiology*; D006801:Humans; D000860:Hypoxia / Q000151:congenital / Q000201:enzymology* / Q000517:prevention & control; D007231:Infant, Newborn; D007234:Infant, Premature / Q000502:physiology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects* / Q000502:physiology; D011651:Pulmonary Artery / Q000473:pathology; D012756:Sheep; D013481:Superoxides / Q000032:analysis; D024483:Vitamin K 3 / Q000494:pharmacology
D000098:Acetophenones; D008070:Lipopolysaccharides; D013481:Superoxides; D024483:Vitamin K 3; C056165:acetovanillone