NO inhibits hyperoxia-induced NF-κB activation in neonatal pulmonary microvascular endothelial cells.
Inhaled NO (iNO) may be protective against hyperoxic injury in the premature lung, but the mechanism is unknown. We hypothesized that NO would prevent hyperoxia-induced nuclear factor kappa B (NF-κB) activation in neonatal pulmonary microvascular endothelial cells [human pulmonary microvascular endothelial cell (HPMEC)] and prevent the up-regulation of target genes. After hyperoxic exposure (O2 >95%), nuclear NF-κB consensus sequence binding increased and was associated with IκBα degradation. Both of these findings were prevented by exposure to NO. Furthermore, intracellular adhesion molecule (ICAM)-1 mRNA and protein levels increased in cells exposed to hyperoxia, an effect abrogated by NO. To evaluate the potentially toxic effect of NO plus hyperoxia, cell viability and proliferation were assessed. Cells exposed to NO plus hyperoxia demonstrated improved survival as measured by trypan blue exclusion when compared with cells exposed to hyperoxia alone. These differences in cell death could not be attributed to apoptosis measured by caspase-3 activity. Finally, cellular proliferation inhibited by hyperoxia was rescued by concurrent exposure to NO. These data demonstrate that NO prevents hyperoxia-induced NF-κB activation in HPMEC and results in decreased expression of adhesion molecules and decreased cellular toxicity. This may help to explain the protective effects of NO on hyperoxic injury in the developing lung vasculature.
Wright
Clyde J
CJ
Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, USA.
Agboke
Fadeke
F
Chen
Fengming
F
LA
Ping
P
Yang
Guang
G
Dennery
Phyllis A
PA
D053148:Caspase 3 / Q000378:metabolism; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D018496:Hyperoxia / Q000378:metabolism*; D020672:I-kappa B Proteins / Q000378:metabolism; D007231:Infant, Newborn; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D008168:Lung / Q000098:blood supply*; D008833:Microcirculation / Q000502:physiology*; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology
D020672:I-kappa B Proteins; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D018799:Intercellular Adhesion Molecule-1; D000072000:NF-KappaB Inhibitor alpha; D009569:Nitric Oxide; D053148:Caspase 3; D010100:Oxygen
Am J Physiol Lung Cell Mol Physiol. 2009 Mar;296(3):L296-306
19074556
Am J Physiol. 1999 Sep;277(3):L596-605
10484468
Annu Rev Physiol. 1986;48:721-31
3518622
Arch Dis Child Fetal Neonatal Ed. 2006 Jan;91(1):F36-9
16049063
FASEB J. 1995 Jul;9(10):899-909
7542214
Cell Death Differ. 2007 Aug;14(8):1393-7
17464324
J Clin Invest. 2004 Sep;114(5):669-78
15343385
Am J Physiol. 1997 Mar;272(3 Pt 1):L418-25
9124598
Lung. 2002;180(6):301-17
12647232
Pediatrics. 2007 Nov;120(5):1088-99
17974747
Clin Perinatol. 2009 Mar;36(1):43-61
19161864
Proc Natl Acad Sci U S A. 2004 Jun 15;101(24):8945-50
15184672
Pediatr Res. 2006 Mar;59(3):401-6
16492979
Free Radic Biol Med. 2006 Jun 1;40(11):1903-13
16716892
J Clin Invest. 1995 Jul;96(1):60-8
7542286
Am J Physiol Lung Cell Mol Physiol. 2000 Feb;278(2):L245-52
10666107
Methods Enzymol. 2008;436:35-48
18237626
J Biol Chem. 1994 Feb 25;269(8):6185-92
7907090
Biochemistry. 2001 Feb 13;40(6):1688-93
11327828
Am J Physiol Lung Cell Mol Physiol. 2007 Jul;293(1):L35-44
17384081
J Biol Chem. 2007 Oct 19;282(42):30667-72
17720813
Free Radic Biol Med. 2000 May 1;28(9):1317-27
10924851
Lab Invest. 1975 May;32(5):619-35
1092910
J Biol Chem. 1995 Jun 9;270(23):14214-9
7775482
Physiol Rev. 2004 Jul;84(3):731-65
15269335
Biochem Biophys Res Commun. 1999 Sep 7;262(3):685-91
10471386
N Engl J Med. 2007 Nov 8;357(19):1946-55
17989387