Heat shock proteins and macrophage resistance to the toxic effects of nitric oxide.
Nitric oxide (NO) functions as a pathophysiological mediator in mammalian tissues. Activated macrophages produce NO as a non-specific immune response directed against invading bacteria or micro-organisms. The same macrophages that initiate the production of NO also can be toxically affected by NO. Incubation of RAW 264.7 macrophages with lipopolysaccharide (LPS) and/or interferon-gamma (INF-gamma) induced the formation of NO by the activation of a cytokine-inducible NO synthase (NOS). The viability of these macrophages was inversely correlated with the formation of nitrite, a final NO-oxidation product measurable in the incubation medium. The addition of an NOS inhibitor, NG-monomethyl-L-arginine, diminished NO formation and preserved cell viability in a dose- and time-dependent fashion. Treatment of macrophages with ten cycles of non-lethal doses of LPS and INF-gamma, each followed by subculturing of the surviving cells, resulted in cell resistance to the NO toxic insult induced by LPS and INF-gamma. These resistant macrophages showed a 2-fold increase in the expression of the constitutive heat shock protein (HSC 70) which is known to be involved in protecting cells against the action of various metabolic insults. Our results establish a link between cell resistance to the toxic effects of NO, and the expression of heat shock proteins in RAW 264.7 macrophages.
Hirvonen
M R
MR
Department of Toxicology, National Public Health Institute, Kuopio, Finland.
Brüne
B
B
Lapetina
E G
EG
D000818:Animals; D002460:Cell Line; D004351:Drug Resistance; D006360:Heat-Shock Proteins / Q000096:biosynthesis*; D007371:Interferon-gamma / Q000633:toxicity; D008070:Lipopolysaccharides / Q000633:toxicity; D008262:Macrophage Activation; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism*; D009569:Nitric Oxide / Q000633:toxicity*; D009599:Nitroprusside / Q000494:pharmacology; D018384:Oxidative Stress; D051381:Rats; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D006360:Heat-Shock Proteins; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D009599:Nitroprusside; D009569:Nitric Oxide; D007371:Interferon-gamma