Ovariectomized mouse uterotrophic assay of 36 chemicals

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  • Ohta Ryo
    Hatano Research Institute, Food and Drug Safety Center
  • Takagi Atsuya
    Division of cellular and molecular toxicology, Biological Safety Research Center, National Institute of Health Sciences
  • Ohmukai Hideo
    Hatano Research Institute, Food and Drug Safety Center
  • Marumo Hideki
    Hatano Research Institute, Food and Drug Safety Center
  • Ono Atsushi
    Division of cellular and molecular toxicology, Biological Safety Research Center, National Institute of Health Sciences
  • Matsushima Yuko
    Division of cellular and molecular toxicology, Biological Safety Research Center, National Institute of Health Sciences
  • Inoue Tohru
    Division of cellular and molecular toxicology, Biological Safety Research Center, National Institute of Health Sciences
  • Ono Hiroshi
    Hatano Research Institute, Food and Drug Safety Center
  • Kanno Jun
    Division of cellular and molecular toxicology, Biological Safety Research Center, National Institute of Health Sciences

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The concern over endocrine disruptors prompted international establishment of a strategic framework for the identification of the estrogenic compounds. OECD has launched the Conceptual Framework tool box containing various screening and testing methods including the uterotrophic assay. The (anti)estrogenicity of 36 chemicals suspected to be estrogen-receptor interactive by in silico and/or in vitro screening in the Extended Scheme for Endocrine Disruptor Screening and Testing of the Ministry of Health, Labour and Welfare, Japan, were monitored by the uterotrophic assay using C57BL/6J ovariectomized adult female mice after a 7-day exposure by oral gavage (po) and subcutaneous injection (sc). Ethynyl estradiol was used as reference for agonist and antagonist detection. In addition, Bisphenol A (sc) and Genistein (po) were tested for the comparison to rat assays. Among the 36, 2-[Bis(4-hydroxy­phenyl)methyl]benzylalcohol, 2,2’,4,4’-Tetrahydroxybenzophenone, 2,4-Dihydroxybenzophenone, 3,3’,5-Triiodothyroacetic acid, New fuchsin and alpha-Naphtholbenzein, showed both estrogenic agonistic and antagonistic activities; first two showed U-shaped dose-response in antagonistic studies. N,N-Diphenyl-p-phenylenediamine, 2,2’-Dihydroxy-4,4’-dimethoxybenzophenone, n-Butyl 4-hydroxybenzoate, and Reserpine were agonistic by sc. Benzo [a] pyrene, Benz [a] anthracene, Dibenz [a,h] anthracene, 2-(2H-Benzotriazol-2-yl)-4,6-di(t-pentyl)phenol, Rosemarinic acid, meta-Thymol, 6-Gingerol, Colchicine, Malachite green base, Fenbuconazole, and Lead acetate were antagonistic. The rest, i.e. n-Heptyl 4-hydroxybenzoate, Tetrazolium violet, Pravastatin sodium salt, Physostigmine, salicylate (1:1), Nordihydroguaiaretic acid, o-Cresolphthalein, 1,3-Dinitrobenzene, C.I. Pigment orange, Tetrabromobis­phenol-A, 2-Hydroxy-4-methoxybenzophenone, Ethylparaben, Propyl p-hydroxybenzoate, Kaempferol, 2-(2-Benzotriazolyl)-p-cresol and Phenolphthalein were negative for both effects. Taking together with in silico/ in vitro screening, the result suggested that the ovariectomized mouse uterotrophic bioassay has sufficient performance comparable to rat for the screening of (anti)estrogenicity of various chemicals.

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