Cytotoxic properties of salivary oxidants.
Salivary peroxidase and to a lesser extent myeloperoxidase are present in significant concentrations in saliva and catalyze the oxidation of thiocyanate anion (SCN-) by H2O2 to yield the potent oxidants hypothiocyanous acid (HOSCN) and its conjugate base hypothiocyanite anion (OSCN-). The objective of this study was to characterize the cytotoxic potential of peroxidase-generated HOSCN/OSCN- toward human erythrocytes. We found that HOSCN/OSCN- (0.25 mM) generated by the peroxidase-H2O2-SCN- system caused significant hemolysis at pH 6.0 but not at pH 6.5, 7.0, or 7.4. Erythrocyte hemoglobin (OxyHb) was oxidized to methemoglobin (MetHb) at all pH values tested; however, the rate of MetHb formation was dramatically increased at low pH and was not affected by inosine hexaphosphate, suggesting that hemoglobin was oxidized primarily by HOSCN. Concurrent with oxidation of hemoglobin (Hb), there was a pH-dependent consumption of HOSCN/OSCN- with more of the oxidant consumed at pH 6.0 compared with pH 6.5, 7.0, or 7.4. The enhanced oxidation of Hb at acidic pH was not due simply to increased membrane permeability by the uncharged species (HOSCN), since both erythrocyte lysate Hb and purified Hb were oxidized to the same extent at low pH as were intact erythrocytes. It is concluded that both OSCN- and HOSCN enter human erythrocytes where the protonated oxidant (HOSCN) mediates hemolysis and oxidizes OxyHb to MetHb, whereas both HOSCN and OSCN- oxidize glutathione (GSH). These data suggest that the extracellular pH may play an important role in modulating the cytotoxic properties of salivary oxidants.
Grisham
M B
MB
Department of Physiology and Biophysics, Louisiana State University Medical Center, Shreveport 71130.
Ryan
E M
EM
D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism; D006454:Hemoglobins / Q000378:metabolism*; D006461:Hemolysis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008706:Methemoglobin / Q000378:metabolism*; D008954:Models, Biological; D010084:Oxidation-Reduction; D009195:Peroxidase / Q000378:metabolism*; D010544:Peroxidases / Q000378:metabolism*; D012463:Saliva / Q000201:enzymology*; D013861:Thiocyanates / Q000378:metabolism* / Q000494:pharmacology*
D006454:Hemoglobins; D013861:Thiocyanates; C014607:hypothiocyanite ion; D008706:Methemoglobin; D010544:Peroxidases; D009195:Peroxidase; D005978:Glutathione; C031760:thiocyanate