Increased production of hydroxyl radical by pericentral microsomes compared to periportal microsomes after pyrazole induction of cytochrome P4502E1.
Cytochrome P4502E1 is localized in the pericentral (PC) zone of the liver acinus to a greater extent than in the periportal (PP) zone. After pyrazole treatment, PC microsomes were more active in oxidizing typical substrates of CYP2E1 than PP microsomes and had an increased content of CYP2E1. The ability of PC and PP microsomes from pyrazole-treated rats to interact with iron and generate reactive oxygen species such as the hydroxyl radical (.OH) was evaluated. A sensitive DNA strand cleavage assay was used to detect .OH; supercoiled plasmid DNA is compact but is converted by .OH-induced single strand breaks to the relaxed open circular state. Microsomes from PC hepatocytes of pyrazole-treated rats were several fold more reactive than PP microsomes in promoting NADPH-dependent DNA strand cleavage with a variety of iron catalysts, including ferric-ATP, ferric-histidine, ferric-citrate, ferric ammonium sulfate, and ferric-EDTA. DNA strand cleavage was inhibited by superoxide dismutase, catalase, and .OH scavengers such as DMSO and ethanol. Rates of H2O2 production were higher with the PC microsomes. These results indicate that rates of .OH production are higher with PC microsomes than PP microsomes after pyrazole treatment to induce cytochrome P4502E1 and suggest the possibility that elevated production of reactive oxygen species may play a role in ethanol toxicity to the PC zone of the liver acinus.
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis*; D004790:Enzyme Induction; D017665:Hydroxyl Radical / Q000378:metabolism*; D007502:Iron Chelating Agents / Q000494:pharmacology; D008099:Liver / Q000033:anatomy & histology; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D010957:Plasmids / Q000187:drug effects; D011720:Pyrazoles / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D000975:Antioxidants; D007502:Iron Chelating Agents; D011720:Pyrazoles; D017665:Hydroxyl Radical; C031280:pyrazole; D003577:Cytochrome P-450 Enzyme System