Hypoxia decreases nitric oxide production by lipopolysaccharide stimulated macrophages

Barson, R; Cui, HM; Jin, Y; Liu, YS; Nelin, LD

HERO ID

975872

Reference Type

Journal Article

Subtype

Abstract

Year

2008

Language

English

HERO ID 975872
Material Type Abstract
In Press No
Year 2008
Title Hypoxia decreases nitric oxide production by lipopolysaccharide stimulated macrophages
Authors Barson, R; Cui, HM; Jin, Y; Liu, YS; Nelin, LD
Journal Free Radical Biology and Medicine
Volume 45
Issue Suppl.
Page Numbers S132-S132
Abstract Oxygen is a co-factor in nitric oxide (NO) production from NO synthases (NOS). We hypothesized that hypoxia may limit NO production by inducible NOS (iNOS) in macrophages. to test this hypothesis we studied RAW 264.7 cells grown in either 21% O2/5% CO2 (normoxia) or 1% O2/5% CO2 (hypoxia). After stimulation with lipopolysaccharide (LPS) production of NO after 24 h was lower in the cells incubated in hypoxia than in the cells incubated in normoxia (309 ± 28 vs 106 ± 4 nmol/mg protein, p<0.005). the levels of NO production after 24 h in non-LPS stimulated cells was very low in both hypoxia and normoxia and did not differ between the groups (5 ± 1 vs 3 ± 1, p = 0.23). Despite the lower NO production seen in hypoxia, the LPS-stimulated iNOS protein levels by Western blot were actually greater in the cells incubated in hypoxia than in normoxia (densitometry values normalized to β-actin, 3.1 ± 0.2 normoxia vs 5.0 ± 0.4 hypoxia, p<0.05). LPS stimulated RAW 264.7 cells were then incubated for 24 h in either normoxia or hypoxia, washed, given fresh media, and then incubated for an additional 24 h in either the other condition or continued in the same condition. The LPS-stimulated cells that were incubated for 48 h in hypoxia had significantly lower levels of NO production than did those cells incubated for 48 h in normoxia (84 ± 5 versus 446 ± 77 nmol/mg protein, p<0.01). LPS-stimulated cells that were incubated in normoxia for 24 h followed by hypoxia for 24 h had lower (p<0.05) levels of NO production (105 ± 33 nmol/mg protein), while cells that were incubated for 24 hours in hypoxia followed by 24 h in normoxia had similar levels of NO production (612 ± 50 nmol/mg protein), compared to those cells that were incubated in normoxia for 48 h. the levels of iNOS protein were greatest (p<0.001 compared to the other 3 groups) in the cells incubated for 48 h in hypoxia, and the cells incubated for 24 h of hypoxia followed by 24 h of normoxia had higher (p<0.05) levels of iNOS protein than did cells incubated for 48 h in normoxia. These results suggest that hypoxia decreases NO production in a macrophage cell line stimulated with LPS, but that iNOS protein expression is actually increased by hypoxia. These data may have important implications for host defense when hypoxia is super-imposed on infection.
Wosid WOS:000260867900372
Url http://www.sciencedirect.com/science/article/pii/S089158490800631X
Is Certified Translation No
Dupe Override No
Conference Location Indianapolis, IN
Conference Name Society for Free Radical Biology and Medicine 15th Annual Meeting
Conference Date November 19-23, 2008
Is Public Yes
Language Text English
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