Hyperoxia increases protein mass of 5-lipoxygenase and its activating protein, flap, and leukotriene B(4) output in newborn rat lungs.
In this study, the authors examined in newborn rat lung tissues the release of leukotriene B(4) (LTB(4)) from tissue explants in vitro, the protein expression of the LT-synthesizing enzyme, 5-lipoxygenase (5-LO), and its activating protein (FLAP), and the effects of in vivo hyperoxic exposure (>95% O(2)) on these parameters. Basal LTB(4) output increased from 0.98 ng/mgDNA/30 min at day 1 to 3.3 ng/mgDNA/30 min at day 28 (P <.05). Exposure of rat pups to >95% O(2) from days 1 to 7 and 60% O(2) from days 7 to 28 stimulated a 1.6-fold (P <.05) increase in LTB(4) output, compared to normoxic pups (to 1.6 ng/mgDNA/30 min) by day 1 and on day 7. The calcium ionophore, A23187, caused an increase in LTB(4) output from both exposure groups, but LTB(4) output was consistently greater (P <.05) from hyperoxia-exposed pups. Western immunoblotting of lung tissue showed that 5-LO and FLAP protein mass increased (P <.05) from days 4 to 14. Hyperoxia exposure increased the mass of both proteins (P <.05). Immunohistochemistry localized 5-LO and FLAP mainly to alveolar macrophages on day 14, but some staining was evident in parenchymal tissue. These data show that hyperoxia increases LTB(4) output, as well as protein levels of 5-LO and FLAP, in newborn rat lungs during early postnatal life. Elevated LTB(4) may contribute to the etiology of newborn lung disease.
Hosford
Gayle E
GE
Department of Physiology, The Perinatal Research Centre, The CIHR Group in Perinatal Health and Disease, University of Alberta, Edmonton, Canada. ghosford@ualberta.ca
Koyanagi
Kim S
KS
Leung
Wynne I
WI
Olson
David M
DM
D058927:5-Lipoxygenase-Activating Proteins; D000818:Animals; D000831:Animals, Newborn; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism*; D000001:Calcimycin / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism*; D018496:Hyperoxia / Q000378:metabolism*; D007476:Ionophores / Q000494:pharmacology; D007975:Leukotriene B4 / Q000378:metabolism*; D008168:Lung / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D016676:Macrophages, Alveolar / Q000201:enzymology; D008565:Membrane Proteins / Q000378:metabolism*; D010100:Oxygen / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley
D058927:5-Lipoxygenase-Activating Proteins; C548594:Alox5ap protein, rat; D002352:Carrier Proteins; D007476:Ionophores; D008565:Membrane Proteins; D007975:Leukotriene B4; D000001:Calcimycin; D001094:Arachidonate 5-Lipoxygenase; D010100:Oxygen