Abstract
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.
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Abbreviations
- BPD:
-
bronchopulmonary dysplasia
- BrdU:
-
bromodeoxyuridine
- EMSA:
-
electrophoretic mobility shift assay
- HPMEC:
-
human pulmonary microvascular endothelial cells
- IκBα:
-
inhibitory kappa b alpha
- iNO:
-
inhaled NO
- NF-κB:
-
nuclear factor kappa B
- RA:
-
room air
- ROS:
-
reactive oxygen species
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Acknowledgements
Patrick Fernando provided technical assistance for completion of this article.
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Supported by a Grant K12-HD00850 from the Pediatric Scientist Development Program [C.J.W.], the iNO Therapeutics Advancing Newborn Medicine Fellowship Grant [C.J.W.], the Marshall-Klaus Perinatal Research Award [C.J.W.], and a Grant RO-1 HL-58752 [P.A.D.].
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Wright, C., Agboke, F., Chen, F. et al. NO Inhibits Hyperoxia-Induced NF-κB Activation in Neonatal Pulmonary Microvascular Endothelial Cells. Pediatr Res 68, 484–489 (2010). https://doi.org/10.1203/PDR.0b013e3181f917b0
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DOI: https://doi.org/10.1203/PDR.0b013e3181f917b0
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