Role of nitric oxide and superoxide anion in spontaneous lung chemiluminescence

被引:8
作者
Barnard, ML
Robertson, B
Watts, BP
Turrens, JF
机构
[1] UNIV S ALABAMA, DEPT BIOMED SCI, MOBILE, AL 36688 USA
[2] MICHAEL REESE HOSP & MED CTR, DEPT PULM & CRIT CARE MED, CHICAGO, IL 60616 USA
关键词
endothelial cells; isolated perfused lung; peroxynitrite; nitric oxide synthetase; oxidative stress;
D O I
10.1152/ajplung.1997.272.2.L262
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
Inhibition of nitric oxide (. NO) synthase by nitro-L-arginine (NLA) decreased baseline chemiluminescence in a dose-dependent fashion up to 78% at 300 mu M NLA. This inhibition was prevented by pretreatment with 1 mM arginine. Similarly, addition of superoxide dismutase (SOD; 200 U/ml) to the perfusion buffer inhibited spontaneous light emission by 57%. Addition of NLA after SOD or vice versa did not inhibit light emission any further, suggesting that both . NO and O-2(-). were precursors of the same oxidant. Production of additional extracellular O-2(-) by neutrophils activated with phorbol 12-myristate 13-acetate increased light emission by >200%, but this increase was insensitive to NLA. Increasing the intracellular steady-state O-2(-). concentration by perfusion of control lungs with the Cu and Zn-containing SOD inhibitor diethyldithiocarbamate (1 mM) stimulated light emission up to fourfold, but this spontaneous chemiluminescence was also insensitive to NLA. In experiments using cultured endothelial cells supplemented with extracellular bovine serum albumin (BSA), 5 mu M of the Ca2+ ionophore A-23187 (a stimulant of NO synthase) stimulated chemiluminescence by 40%. This increase was again SOD and NLA sensitive. Addition of NLA after SOD or vice versa did not change light emission. These results suggest that the background chemiluminescence of isolated-perfused intact lungs may result from the constant release of small amounts of O-2(-) and . NO by endothelial cells into the capillary lumen, which in turn react with BSA in the perfusion buffer.
引用
收藏
页码:L262 / L267
页数:6
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