Biochemical mechanisms of hydrogen peroxide- and hypochlorous acid-mediated inhibition of human mononuclear leukocyte functions in vitro: protection and reversal by anti-oxidants.
Both H2O2 (IC50 = 70 microM) and HOCl (IC50 = 8.5 microM) inhibited mitogen-induced MNL proliferation in a dose-dependent manner. This was found to be due to a depletion of intracellular ATP by at least two distinct mechanisms. HOCl and high concentrations (greater than 100 microM) of H2O2 inhibit ATP generation via sulfhydryl group oxidation on the active site of the glyceraldehyde-3-phosphate dehydrogenase (G3PDH) enzyme of the glycolytic pathway. On the other hand, low H2O2 concentrations cause ATP depletion by an activation of the DNA repair enzyme, poly(ADP-ribose)polymerase (pADPRP), leading to consumption of NAD+, an essential cofactor for G3PDH. The anti-oxidants ascorbate and cysteine protected MNL against the anti-proliferative effects of HOCl. Similar results were achieved with the HOCl-mediated inhibition of ATP production and G3PDH activity. However, ascorbate was unable to protect against H2O2-mediated inhibition of MNL functions, while cysteine protected against the inhibitory effects on ATP production and G3PDH activity, induced by this oxidant.
Smit
M J
MJ
Department of Immunology, Faculty of Medicine, University of Pretoria, Republic of South Africa.
Anderson
R
R
D000255:Adenosine Triphosphate / Q000378:metabolism; D000975:Antioxidants / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D005954:Glucosephosphate Dehydrogenase / Q000494:pharmacology; D006019:Glycolysis / Q000187:drug effects; D006801:Humans; D006861:Hydrogen Peroxide / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D006997:Hypochlorous Acid / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D066298:In Vitro Techniques; D007773:Lactates / Q000097:blood; D007963:Leukocytes, Mononuclear / Q000187:drug effects* / Q000201:enzymology; D008081:Liposomes; D008934:Mitogens / Q000494:pharmacology; D009243:NAD / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000494:pharmacology
D000975:Antioxidants; D007773:Lactates; D008081:Liposomes; D008934:Mitogens; D009243:NAD; D006997:Hypochlorous Acid; D000255:Adenosine Triphosphate; D006861:Hydrogen Peroxide; D005954:Glucosephosphate Dehydrogenase; D011065:Poly(ADP-ribose) Polymerases