Modification of the xanthine-converting enzyme of perfused rat heart during ischemia and oxidative stress.
The reversible and irreversible conversion of xanthine dehydrogenase to xanthine oxidase during ischemia/reperfusion and oxidative stress induced by hydrogen peroxide or diamide and its relationship with glutathione and protein SH groups were studied. The direct spectrophotometric measurement of the various forms of the xanthine-converting enzyme indicates that, in the fresh rat heart or after normoxic perfusion, there always is a basal level of 80% xanthine dehydrogenase and 20% of xanthine oxidase (15% irreversible and 5% reversible) that could contribute to the background production of free radicals. There is no significant increase of irreversible xanthine oxidase during ischemia nor during reperfusion. After global ischemia the reversible oxidase shows almost no increase while, when ischemia is followed by reperfusion, there is a limited increase (less then 9%) of the reversible xanthine oxidase. In the latter conditions there is a decrease of glutathione and of SH groups of about 70% and 25%, respectively. Perfusion for 1 h with oxidizing agents like hydrogen peroxide (60 microM) or diamide (100 microM) determines a marked conversion of xanthine dehydrogenase to reversible xanthine oxidase of about 40% and 60%, respectively; this oxidase activity partially reconverts to the dehydrogenase after withdrawing the oxidizing agents from the perfusion medium. The level of irreversible xanthine oxidase remains unchanged in all the conditions tested. Both hydrogen peroxide and diamide induce a strong decrease in SH groups and depletion of glutathione. The xanthine dehydrogenase----xanthine oxidase conversion thus appears to be sensitive to the redox state of thiol groups.
Bindoli
A
A
Centro Studio Fisiologia Mitochondriale, C. N. R., Padova, Italy.
Cavallini
L
L
Rigobello
M P
MP
Coassin
M
M
Di Lisa
F
F
D000818:Animals; D003326:Coronary Circulation; D005609:Free Radicals; D005978:Glutathione / Q000378:metabolism; D066298:In Vitro Techniques; D007511:Ischemia / Q000201:enzymology*; D007658:Ketone Oxidoreductases / Q000378:metabolism*; D009206:Myocardium / Q000201:enzymology*; D010477:Perfusion; D051381:Rats; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014968:Xanthine Dehydrogenase / Q000378:metabolism*; D014969:Xanthine Oxidase / Q000378:metabolism*
D005609:Free Radicals; D013438:Sulfhydryl Compounds; D014968:Xanthine Dehydrogenase; D014969:Xanthine Oxidase; D007658:Ketone Oxidoreductases; D005978:Glutathione