title: AHTN and HHCB show weak estrogenic--but no uterotrophic activity.
abstract: The ubiquitous presence of the polycyclic musks AHTN (6-acetyl-1,1,2,4,4,7-hexamethyltetraline) and HHCB (1,2,4,6,7,8-hexahydro-4,6,6,7,8-hexamethylcyclopenta-gamma-2-b enzopyreen) in surface waters and their identification in human milk fat together with their polycyclic nature, which makes them potential candidates for interference with estrogen receptors, prompted us to assess these compounds for their potential estrogenic effects. We therefore investigated the effects of AHTN and HHCB in ERalpha- and ERbeta-dependent gene transcription assays with Human Embryonal Kidney 293 (HEK293) cells, which have proven to be very suitable to estimate the estrogenic activity of compounds with low binding activity (Kuiper, G.G., Lemmen, J.G., Carlsson, B., Corton, J.C., Safe, S.H., Van der Saag, P.T., Van der Burg, B., Gustafsson, J.A., 1998. Interaction of estrogenic chemicals and phytoestrogens with estrogen receptor beta. Endocrinology 139, 4252-4264). Both AHTN and HHCB were found to induce a slight but dose-dependent stimulation of transcriptional activity in the transiently ERalpha transfected HEK293 cells. This weak estrogenic response was not observed in the ERbeta transiently transfected cells. However, these cells were less responsive to estradiol than the ERalpha transfected HEK293 cells. Also, no significant increase in transcriptional activity was observed in HEK293 cell lines, permanently expressing the same estrogen-responsive reporter gene construct and either ERalpha or ERbeta. In the classical uterine weight assay performed in juvenile Balb/c mice, no uterotrophic activity of AHTN and HHCB was noted at relatively high dietary exposure levels up to 50 and 300 ppm, respectively, at which levels an increase in liver weight was evident. Also the vitellogenin production by carp hepatocytes, a sensitive marker of estrogenic activity, was not affected by these two fragrance materials (Smeets, J.M.W., Rouhani Rankouhi, T., Nichols, K.M., Komen, H., Kaminsky, N.E., Giesy, J.P., Van den Berg, M., 1999. In vitro vitellogenin production by carp (Cyprimus carpio) hepatocytes as a screening method for determining (anti-) estrogenic activity of xenobiotics. Toxicol. Appl. Pharmacol., 157, 68-76). Therefore it is concluded that these compounds have very weak estrogenic potency, too weak to induce estrogenic effects in wildlife species or humans at the current levels of exposure. These results give further support to the promiscuity of estrogen receptors.
authors.affiliation: Toxicology Research Institute, Utrecht University, The Netherlands. w.seinen@ritox.vet.uu.nl
authors.forename: W
authors.initials: W
authors.lastname: Seinen
authors.forename: J G
authors.initials: JG
authors.lastname: Lemmen
authors.forename: R H
authors.initials: RH
authors.lastname: Pieters
authors.forename: E M
authors.initials: EM
authors.lastname: Verbruggen
authors.forename: B
authors.initials: B
authors.lastname: van der Burg
mesh_terms: D000818:Animals; D001578:Benzopyrans / Q000633:toxicity*; D002460:Cell Line; D004967:Estrogens / Q000633:toxicity*; D005229:Fatty Acids, Monounsaturated / Q000633:toxicity*; D005260:Female; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D009281:Naphthalenes / Q000633:toxicity*; D010476:Perfume / Q000633:toxicity*; D011960:Receptors, Estrogen / Q000187:drug effects / Q000502:physiology; D014158:Transcription, Genetic / Q000187:drug effects; D014599:Uterus / Q000187:drug effects*
chemical_list: D001578:Benzopyrans; D004967:Estrogens; D005229:Fatty Acids, Monounsaturated; D009281:Naphthalenes; D010476:Perfume; D011960:Receptors, Estrogen; C008563:musk