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J.Health Sci., 49(6), 481-491, 2003

Estrogenic Activity of a Diet to Estrogen Receptors -alpha and -beta in an Experimental Animal

Maki Miyahara,a Hiroshi Ishibashi,a Makiko Inudo,a Haruka Nishijima,a Taisen Iguchi,b Louis Joseph Guillette Jr.,c and Koji Arizono*, a

aFaculty of Environmental and Symbiotic Sciences, Prefectural University of Kumamoto, 3-1-100 Tsukide, Kumamoto 862-8502, Japan, bOkazaki National Research Institutes Center for Integrative Bioscience Bioenvironmental Science, 38 Nishigonaka, Myodaiji, Okazaki, Aichi 444-8585, Japan, and cDepartment of Zoology, University of Florida, 223 Bartram Hall PO Box 118525, Gainesville, FL 32611 U.S.A.

Phytoestrogens, such as daidzein and genistein in plants are suspected as endocrine-disrupting chemicals (EDCs), because their chemical structures are similar to natural or synthetic estrogens, and have estrogenic activity in vitro and in vivo. An EDC study was carried out on the diets of various animals in vivo. However, many of these diets include phytoestrogens and may already possess estrogenic activity. In this study, we evaluated the estrogenic activity of phytoestrogens (such as daidzin, genistin, daidzein, genistein, coumestrol and equol) and the feed diets for experimental animals, such as fish, amphibians and reptiles, towards human estrogen receptors alpha (hER-alpha) and beta (hER-beta), and the genistein and daidzein content in these diets from HPLC analysis. Coumestrol showed the highest estrogenic activity for hER-alpha and -beta in the -S9 test. Equol showed the highest estrogenic activity for hER-alpha in the +S9 test. The estrogenic activities of coumestrol, equol and genistein were approximately one hundred to two thousand times higher than that of daidzein. Many of these compounds showed higher compatibility with hER-beta than with hER-alpha. A diet for fish from soybean was indicated to contain the highest amounts of genistein and daidzein. Moreover, this fish diet had the highest estrogenic activity for hER-alpha and -beta. The estrogenic activity was found with hydrolysis by beta-glucuronidase, showing higher compatibility with hER-beta than with hER-alpha. In addition, correlation between the contents of genistein and estrogenic activities in the diets was found, with the exception of part of the diet. Therefore, this indicates that the genistein content contributes to the estrogenic activity of the diets. These results suggest that in vivo estrogenic activity might be caused by the diet provided to an experimental animal, indicating the necessity for more careful selection of the feeding diet and measurement of estrogenic substances when performing an in vivo screening assay for EDCs.