Intracellular high energy metabolite depletion and cell membrane injury with antioxidant enzymes during oxidant exposure in vitro.
We compared oxidant-induced intracellular adenine nucleotide catabolism and cell membrane injury in 4 different human cell types. Responses to oxidant exposure were correlated with endogenous antioxidant enzyme activities in these cells. Blood monocytes, amniotic fibroblasts, umbilical vein endothelial cells in primary culture, and transformed bronchial epithelial cells (BEAS 2B) were exposed to 0.1-5 mM hydrogen peroxide (H2O2) for 4 h. Some experiments were conducted in cells pretreated with 3-amino 1:2,4-triazole (ATZ) to inactivate catalase or with 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) to inactivate glutathione (GSH) reductase. Depletion of adenine nucleotides and accumulation of their catabolic products (hypoxanthine, xanthine and uric acid) occurred to varying extent, monocytes being the most resistant. There was a mutual relationship between catalase and GSH reductase activities and maintenance of cellular adenine nucleotide levels during H2O2 exposure. GSH reductase inhibition rendered BEAS 2B cells susceptible to lytic injury by H2O2, assessed by release of lactate dehydrogenase and intact nucleotides into the medium, there was no correlation between these markers of such injury and endogenous antioxidant enzymes. We conclude that adenine nucleotide depletion and nucleotide catabolite accumulation relate closely with the antioxidant enzyme activities, whereas the lack of a similar correlation between the enzyme levels and markers of lytic cell injury suggest that intracellular antioxidant enzymes do not protect cells from membrane damage due to extracellular oxidants.
Aalto
K
K
University of Helsinki, Department of Pediatrics, Finland.
Raivio
K O
KO
Pietarinen
P
P
Kinnula
V L
VL
D000227:Adenine Nucleotides / Q000378:metabolism*; D000975:Antioxidants / Q000378:metabolism; D002330:Carmustine / Q000494:pharmacology; D002374:Catalase / Q000378:metabolism*; D002461:Cell Line, Transformed; D002462:Cell Membrane / Q000187:drug effects*; D002478:Cells, Cultured; D004734:Energy Metabolism; D005980:Glutathione Reductase / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D016877:Oxidants / Q000494:pharmacology*; D015215:Zidovudine / Q000494:pharmacology
D000227:Adenine Nucleotides; D000975:Antioxidants; D016877:Oxidants; D015215:Zidovudine; D006861:Hydrogen Peroxide; D007770:L-Lactate Dehydrogenase; D002374:Catalase; D005980:Glutathione Reductase; D002330:Carmustine