title: Evaluation for a mutagenicity of aristolochic acid by Pig-a and PIGRET assays in rats.
abstract: The Pig-a assay, which uses the endogenous phosphatidylinositol glycan, class A gene (Pig-a) as a reporter of mutation, has been developed as a method for evaluating in vivo mutagenicity. Pig-a gene mutation can be detected by identifying the presence of CD59, the glycosylphosphatidylinositol anchor protein, on the surface of erythrocytes (RBC Pig-a assay) and reticulocytes (PIGRET assay). The International Workshop on Genotoxicity Testing (IWGT) showed the usefulness of the RBC Pig-a assay through the evaluation of several compounds. Aristolochic acid (AA), one of the evaluated compounds in the IWGT workgroup, is a carcinogenic plant toxin that is a relatively strong gene mutagen both in vitro and in vivo, but a weak inducer of micronuclei in vivo. In the present study, we examined the mutagenicity of AA in the peripheral blood of rats treated orally with a single dose of AA using Pig-a assays. Furthermore, we evaluated the advantages of the PIGRET assay compared with the RBC Pig-a assay. The results showed that a statistically significant increase in mutant frequency of the Pig-a gene was detected at day 28 by the RBC Pig-a assay, and at days 7, 14 and 28 by the PIGRET assay. In addition, the mutant frequency by the PIGRET assay was higher than that by the RBC Pig-a assay. These results indicate that the mutagenicity of AA can be detected using the Pig-a assays, as reported by the IWGT, and the PIGRET assay can detect Pig-a mutants at an early time point compared with the RBC Pig-a assay.
authors.affiliation: Pharmacokinetics and Safety Department, Drug Research Center, Kaken Pharmaceutical Co., LTD., 301, Gensuke, Fujieda-shi, Shizuoka 426-8646, Japan. Electronic address: koyama_naomi@kaken.co.jp.
authors.forename: Naomi
authors.initials: N
authors.lastname: Koyama
authors.affiliation: Pharmacokinetics and Safety Department, Drug Research Center, Kaken Pharmaceutical Co., LTD., 301, Gensuke, Fujieda-shi, Shizuoka 426-8646, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Yonezawa
authors.affiliation: Pharmacokinetics and Safety Department, Drug Research Center, Kaken Pharmaceutical Co., LTD., 301, Gensuke, Fujieda-shi, Shizuoka 426-8646, Japan.
authors.forename: Michi
authors.initials: M
authors.lastname: Nakamura
authors.affiliation: Pharmacokinetics and Safety Department, Drug Research Center, Kaken Pharmaceutical Co., LTD., 301, Gensuke, Fujieda-shi, Shizuoka 426-8646, Japan.
authors.forename: Hisakazu
authors.initials: H
authors.lastname: Sanada
mesh_terms: D000818:Animals; D034341:Aristolochic Acids / Q000633:toxicity*; D001835:Body Weight; D004912:Erythrocytes / Q000187:drug effects*; D008297:Male; D008565:Membrane Proteins / Q000235:genetics*; D009152:Mutagenicity Tests / Q000379:methods*; D009153:Mutagens / Q000633:toxicity*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012156:Reticulocytes / Q000187:drug effects*
chemical_list: D034341:Aristolochic Acids; D008565:Membrane Proteins; D009153:Mutagens; C080086:phosphatidylinositol glycan-class A protein; C000228:aristolochic acid I
keywords: Aristolochic acid; PIGRET assay; RBC Pig-a assay