Inactivation of alcohol dehydrogenase by piroxicam-derived radicals.
Alcohol dehydrogenase (ADH) was used as a marker molecule to clarify the mechanism of gastric mucosal damage as a side effect of using piroxicam. Piroxicam inactivated ADH during interaction of ADH with horseradish peroxidase and H2O2 (HRP-H2O2). The ADH was more easily inactivated under aerobic than anaerobic conditions, indicating participation by oxygen. Superoxide dismutase, but not hydroxyl radical scavengers, inhibited inactivation of ADH, indicating participation by superoxide. Sulfhydryl (SH) groups in ADH were lost during incubation of piroxicam with HRP-H2O2. Adding reduced glutathione (GSH) efficiently blocked ADH inactivation. Other SH enzymes, including creatine kinase and glyceraldehyde-3-phosphate dehydrogenase, were also inactivated by piroxicam with HRP-H2O2. Thus SH groups in the enzymes seem vulnerable to piroxicam activated by HRP-H2O2. Spectral change in piroxicam was caused by HRP-H2O2. ESR signals of glutathionyl radicals occurred during incubation of piroxicam with HRP-H2O2 in the presence of GSH. Under anaerobic conditions, glutathionyl radical formation increased. Thus piroxicam free radicals interact with GSH to produce glutathionyl radicals. Piroxicam peroxyl radicals or superoxide, or both, seem to inactivate ADH. Superoxide may be produced through interaction of peroxyl radicals with H2O2. Thus superoxide dismutase may inhibit inactivation of ADH through reducing piroxicam peroxyl radicals or blocking interaction of SH groups with O2 , or both. Other oxicam derivatives, including isoxicam, tenoxicam and meloxicam, induced ADH inactivation in the presence of HRP-H2O2.
Muraoka
Sanae
S
Department of Biology, Hokkaido College of Pharmacy, Katsuraoka-cho 7-1, Otaru 0470264, Japan.
Miura
Toshiaki
T
D000426:Alcohol Dehydrogenase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D003402:Creatine Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism; D005609:Free Radicals / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D005978:Glutathione / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000737:chemistry / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D010894:Piroxicam / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013997:Time Factors
D005609:Free Radicals; D013438:Sulfhydryl Compounds; D010894:Piroxicam; D006861:Hydrogen Peroxide; D000426:Alcohol Dehydrogenase; D006735:Horseradish Peroxidase; D003402:Creatine Kinase; D005978:Glutathione