Comparison of the Hershberger assay and androgen receptor binding assay of twelve chemicals.

We performed the Hershberger assay of 12 chemicals based on the OECD draft protocol. The chemicals tested by the Hershberger assay were phthalic acid di-n-hexyl ester, phthalic acid di-n-amyl ester, phthalic acid di-n-propyl ester, diethylstilbestrol, 17beta-estradiol, tamoxifen, 5alpha-dihydrotestosterone, dichlorodiphenyldichloroethane, cyproterone acetate, 6alpha-methyl-17alpha-hydroxy-progesterone, atrazine, and spironolactone. Phthalic acid di-n-hexyl ester, phthalic acid di-n-amyl ester, and phthalic acid di-n-propyl ester are phthalates; diethylstilbestrol and 17beta-estradiol are estrogenic chemicals; tamoxifen is partial estrogen receptor antagonist with mainly estrogenic properties; 5alpha-dihydrotestosterone is an androgen derivatives; dichlorodiphenyldichloroethane is a reference androgen antagonistic chemical; cyproterone acetate, 6alpha-methyl-17alpha-hydroxy-progesterone, and spironolactone have an androgenic steroid structure and are known as androgen antagonistic chemicals; and atrazine is a reference endocrine disruptor. We also subjected these chemicals to the receptor binding assay for androgen. A clear androgen agonistic effect was detected in 5alpha-dihydrotestosterone, and an androgen antagonistic effect was observed in five chemicals: cyproterone acetate, spironolactone, 6alpha-methyl-17alpha-hydroxy-progesterone, phthalic acid di-n-amyl ester, and dichlorodiphenyldichloroethane. By contrast, diethylstilbestrol, 17beta-estradiol, tamoxifen, 5alpha-dihydrotestosterone, dichlorodiphenyldichloroethane, cyproterone acetate, 6alpha-methyl-17alpha-hydroxy-progesterone, and spironolactone were positive in the receptor binding assay for androgen. Three estrogenic chemicals, diethylstilbestrol, 17beta-estradiol, and tamoxifen, were negative in the Hershberger assay with receptor binding affinity. On the other hand, the Hershberger assays of three phthalates were performed at the same dosages, and the results showed androgen antagonistic affinity only in the assay of phthalic acid di-n-amyl ester without receptor binding affinity.

[1]  M. Takatsuki,et al.  Immature rat uterotrophic assay of 18 chemicals and Hershberger assay of 30 chemicals. , 2003, Toxicology.

[2]  Andrew S Friedmann,et al.  Atrazine inhibition of testosterone production in rat males following peripubertal exposure. , 2002, Reproductive toxicology.

[3]  P. Wainman,et al.  THE EFFECTS OF CASTRATION AND TESTOSTERONE PROPIONATE ON THE STRIATED PERINEAL MUSCULATURE IN THE RAT , 1941 .

[4]  G. Gordan,et al.  The levator ani muscle of the rat as an index of myotrophic activity of steroidal hormones. , 1950, The Journal of pharmacology and experimental therapeutics.

[5]  V. Korenchevsky.  The assay of testicular hormone preparations. , 1932, The Biochemical journal.

[6]  R. Cooper,et al.  The effects of atrazine on female wistar rats: an evaluation of the protocol for assessing pubertal development and thyroid function. , 2000, Toxicological sciences : an official journal of the Society of Toxicology.

[7]  R. Sharpe,et al.  Neonatal exposure to potent and environmental oestrogens and abnormalities of the male reproductive system in the rat: evidence for importance of the androgen–oestrogen balance and assessment of the relevance to man , 2001 .

[8]  J. Mornon,et al.  Steroid hormone receptors and pharmacology. , 1980, Journal of steroid biochemistry.

[9]  A. Panzeri,et al.  Endocrine properties of the testosterone 5α-reductase inhibitor turosteride (FCE 26073) , 1994, The Journal of Steroid Biochemistry and Molecular Biology.

[10]  V. Korenchevsky,et al.  Changes produced by testicular hormone in normal and in castrated rats. , 1933, The Biochemical journal.

[11]  L. Andersson,et al.  A role for estrogen receptor β in the regulation of growth of the ventral prostate , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[12]  P. Foster,et al.  Disruption of androgen-regulated male reproductive development by di(n-butyl) phthalate during late gestation in rats is different from flutamide. , 1999, Toxicology and applied pharmacology.

[13]  J. Freud,et al.  Differences between Male Hormone Extracts from Urine and from Testes , 1935, Nature.

[14]  Edward F. Orlando,et al.  Effects of environmental antiandrogens on reproductive development in experimental animals , 2001 .

[15]  V. Korenchevsky,et al.  The response of castrated male rats to the injection of testicular hormone. , 1932, The Biochemical journal.

[16]  E. Bülbring,et al.  The estimation of œstrin and of male hormone in oily solution , 1935, The Journal of physiology.

[17]  D. Vigo,et al.  Effects of mepartricin on estradiol and testosterone serum levels and on prostatic estrogen, androgen and adrenergic receptor concentrations in adult rats. , 2001, Pharmacological research.

[18]  A. S. Parkes,et al.  Comparative activities of compounds of the androsterone-testosterone series. , 1936, The Biochemical journal.

[19]  R. K. Meyer,et al.  Myotrophic Activity of 19-Nortestosterone and Other Steroids Determined by Modified Levator Ani Muscle Method.∗ , 1953, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[20]  G. Gordan,et al.  Testosterone and tissue respiration of the castrate male rat with a possible test for myotrophic activity. , 1949, Endocrinology.

[21]  F. Labrie,et al.  The pure antiandrogen ru 23908 (anandron®), a candidate of choice for the combined antihormonal treatment of prostatic cancer: A review , 1984, The Prostate.

[22]  A. T. Glen,et al.  Receptor binding and biological activity of steroidal and nonsteroidal antiandrogens. , 1981, Journal of steroid biochemistry.

[23]  R. Neri,et al.  On the Mechanism of the Anti-androgenic Action ofFlutamide (α-α-α-Trifluoro-2-methyl-4'-nitro-m-propionotoluidide)in the Rat , 1974 .