title: Studies of the toxicological potential of tripeptides (L-valyl-L-prolyl-L-proline and L-isoleucyl-L-prolyl-L-proline): VII. Micronucleus test of tripeptides-containing casein hydrolysate and Lactobacillus helveticus-fermented milk powders in rats and mice.
abstract: The objective of these in vivo experiments was to assess the mutagenic potential of tripeptides as reflected by the ability of the test compound to induce the formation of micronuclei in mouse polychromatic erythrocytes. The test agents used in these experiments were (1) powdered Aspergillus oryzae protease casein hydrolysate (CH) and (2) powdered Lactobacillus helveticus-fermented milk (FM). Both test agents contain two tripeptides, L-valyl-L-prolyl-L-proline (VPP) and L-isoleucyl-L-prolyl-L-proline (IPP). Male Sprague-Dawley rats (five per group) were administered doses of 0, 500, 1000, or 2000 mg (0, 3, 6, or 12 mg VPP plus IPP)/kg body weight (BW)/day CH by oral gavage for 2 days. Male CD-1 mice (six per group) received a single oral gavage dose of 0, 500, 1000, or 2000 mg (0, 0.8, 1.6 or 3.3 mg VPP plus IPP)/kg BW of FM. Positive-control agents were cyclophosphamide (10 mg/kg, intraperitoneal [i.p.]) in rats and mitocycin C (2 mg/kg, i.p.) in mice. Twenty-four hours after the second dose of CH, or the dose of cyclophosphamide to rats, or FM or mitocycin C to mice, bone marrow cells were fixed and examined for the presence of polychromatic erythrocytes (PCEs) and the presence or absence of micronucleated PCEs (MNPCEs). Administration of CH to rats and FM to mice produced neither changes in body weights nor signs of systemic toxicity. Similarly, neither CH nor FM caused statistically significant variations in the incidences of either PCEs or MNPCEs. Both positive-control agents caused unequivocal increases in the incidence of MNPCEs and cyclophosphamide significantly reduced the percent of rat erythrocytes appearing as PCEs. The results of these micronucleus assays conducted with either powdered CH or FM in rats and mice, respectively, show that neither form of the tripeptides possesses the potential to induce micronuclei formation in these rodent species.
authors.affiliation: Product Development Laboratory, Calpis Co., Ltd., 11-10, 5-chome, Fuchinobe, Sagamihara, Kanagawa 229-0006, Japan. keiichi.matsuura@calpis.co.jp
authors.forename: Keiichi
authors.initials: K
authors.lastname: Matsuura
authors.forename: John H
authors.initials: JH
authors.lastname: Mennear
authors.forename: Masafumi
authors.initials: M
authors.lastname: Maeno
authors.forename: Bruce K
authors.initials: BK
authors.lastname: Bernard
mesh_terms: D000284:Administration, Oral; D000818:Animals; D001236:Aspergillus oryzae / Q000737:chemistry / Q000201:enzymology; D001854:Bone Marrow Cells / Q000187:drug effects; D002364:Caseins / Q000008:administration & dosage / Q000737:chemistry; D043302:Cultured Milk Products / Q000737:chemistry*; D003520:Cyclophosphamide / Q000008:administration & dosage / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D004900:Erythroblasts / Q000187:drug effects; D052198:Lactobacillus helveticus / Q000254:growth & development / Q000378:metabolism*; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D015162:Micronucleus Tests / Q000379:methods; D016685:Mitomycin / Q000008:administration & dosage / Q000633:toxicity; D009842:Oligopeptides / Q000008:administration & dosage* / Q000737:chemistry / Q000633:toxicity; D010447:Peptide Hydrolases / Q000008:administration & dosage / Q000737:chemistry; D011208:Powders; D051381:Rats; D017207:Rats, Sprague-Dawley; D018675:Toxicity Tests / Q000379:methods
chemical_list: D002364:Caseins; D009842:Oligopeptides; D011208:Powders; C489032:isoleucyl-prolyl-proline; C489031:valyl-prolyl-proline; D016685:Mitomycin; C475729:casein hydrolysate; D003520:Cyclophosphamide; D010447:Peptide Hydrolases