Aristolactam I a metabolite of aristolochic acid I upon activation forms an adduct found in DNA of patients with Chinese herbs nephropathy.
Aristolochic acid (AA) a naturally occuring nephrotoxin and carcinogen is implicated in a unique type of renal fibrosis, designated Chinese herbs nephropathy (CHN). We identified AA-specific DNA adducts in kidneys and in a ureter obtained from CHN patients after renal transplantation. AA is a plant extract of aristolochia species containing AA I as the major component. Aristolactams are the principal detoxication metabolites of AA, which were detected in urine and faeces from animals and humans. They are activated by cytochrome P450 (P450) and peroxidase to form DNA adducts. Using the 32P-postlabelling assay we investigated the formation of DNA adducts by aristolactam I in these two activation systems. A combination of two independent chromatographic systems (ion-exchange chromatography TLC and reversed-phase HPLC) with reference compounds was used for the identification of adducts. Aristolactam I activated by peroxidase led to the formation of several adducts. Two major adducts were identical to adducts previously observed in vivo. 7-(deoxyguanosin-N2-yl)aristolactam I (dG-AAI) and 7-(deoxyadenosin-N6-yl)aristolactam I (dA-AAI) were formed in DNA during the peroxidase-mediated one-electron oxidation of aristolactam I. Aristolactam I activated by P450 led to one major adduct and four minor ones. Beside the principal AA-DNA adducts identified recently in the ureter of one patient with CHN, an additional minor adduct was detected, which was found to have indistinguishable chromatographic properties on TLC and HPLC from the major adduct formed from aristolactam I by P450 activation. Thus, this minor AA-adduct might be evolved from the AAI detoxication metabolite (aristolactam I) by P450 activation. These results indicate a potential carcinogenic effect of aristolactam I in humans.
Stiborová
M
M
Department of Biochemistry, Charles University, Prague, The Czech Republic.
Frei
E
E
Breuer
A
A
Bieler
C A
CA
Schmeiser
H H
HH
D000818:Animals; D034341:Aristolochic Acids; D002851:Chromatography, High Pressure Liquid; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D018736:DNA Adducts / Q000032:analysis / Q000378:metabolism*; D004149:Dioxoles / Q000032:analysis / Q000378:metabolism*; D004365:Drugs, Chinese Herbal / Q000009:adverse effects*; D005355:Fibrosis; D006735:Horseradish Peroxidase / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007211:Indoles / Q000032:analysis / Q000378:metabolism*; D007674:Kidney Diseases / Q000139:chemically induced*; D008297:Male; D010616:Phenanthrenes / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014513:Ureter / Q000737:chemistry
D034341:Aristolochic Acids; D018736:DNA Adducts; D004149:Dioxoles; D004365:Drugs, Chinese Herbal; D007211:Indoles; D010616:Phenanthrenes; C121135:aristolactam I; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; C000228:aristolochic acid I; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase