DNA damage in human mononuclear cells induced by bacterial endotoxin.
Damage to nuclear DNA in human peripheral blood mononuclear cells was studied after in vitro treatment with bacterial endotoxin by alkaline comet assay. It was found that LPS induced DNA damage as soon as over the first 30 min of incubation, while by the 4th hour of incubation DNA damage was found in more than 95% cells. Exogenous superoxide dismutase completely protected DNA, which suggests that superoxide radical is the primary extracellular damaging agent. Polyphenol antioxidant (water-soluble lignin) and specific NADPH oxidase inhibitor (diphenyleneiodonium chloride) also produced a protective effect. Our results show that LPS-activated mononuclear cells can be used ex vivo as a convenient and adequate experimental system for evaluation of the efficiency of various substances in protection of lymphocyte DNA from the damaging effect of reactive oxygen species of LPS-stimulated monocytes.
Glukhov
I L
IL
G. K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino, Russia. glukhovil@rambler.ru
Sirota
N P
NP
Kuznetsova
E A
EA
D020552:Comet Assay; D004249:DNA Damage; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000187:drug effects*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors; D009861:Onium Compounds / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000494:pharmacology; D013481:Superoxides / Q000494:pharmacology
D008070:Lipopolysaccharides; D009861:Onium Compounds; D013481:Superoxides; C007517:diphenyleneiodonium; D013482:Superoxide Dismutase; D019255:NADPH Oxidases