Identification of estrogenic and antiestrogenic activities of respirable diesel exhaust particles by bioassay-directed fractionation
- 61 Downloads
Bioassay-directed fractionation was performed to identify causative chemical groups of DEPs with estrogenic and antiestrogenic activities. Bioassay-directed fractionation consists of a cell bioassay (E-SCREEN) in conjunction with acid-base partitoning (F1 and F2) and silica gel column fractionation of neutral fractions (F3-F7). Crude extract (CE) of DEPs in dichloromethane (DCM) exhibited both estrogenic and antiestrogenic activity. Estrogenic activity of CE and some fractions (F1, F2, F3, F5 and F6) was induced through estrogen receptor (ER)-mediated pathways. In particular, the acid polar fraction (F2) of DEPs, which contains phenols, induced high levels of estrogenic activity compared to other fractions. The estrogenic activity of F2 (610.80 pg-bio-EEQ/g-DEPs) was higher than that of the total estrogenic activity of CE (222.22 pg-bio-EEQ/g-DEPs). This result indicates that the estrogenic activity induced by causative estrogenic fraction (F2) may be antagonized by unidentified chemicals in DEPs. On the other hand, non-polar fractions (F3 and F4) of DEPs include aliphatic and chlorinated hydrocarbon, polyaromatic hydrocarbons, and their alkyl derivatives, which play an important role in the antiestrogenic activity of DEPs. In particular, F4, which contains PAH and its derivatives, showed the highest antiestrogenic activity. Since in our previous study, dibenzo(a, h)anthracene and chrysene were identified in F4, and these chemicals have antiestrogenic activity, we assume that these chemicals are the major causative chemicals with antiestrogenic activity in DEPs. In contrast to the estrogenic activity of DEPs, antiestrogenic activity of CE was stronger than that of antiestrogenic fractions (F3 and F4) at non-cytotoxic concentrations, indicating that additive or synergistic effects by unidentified chemicals contained in DEPs occurred.
Key wordsDiesel exhaust particles (DEPs) Bioassay-directed fractionation E-screen assay bio-EEQ Anti-/estrogenic activity
Unable to display preview. Download preview PDF.
- Gozgit, J. M., Nestor, K. M., Fasco, M. J., Pentecost, B. T., and Arcaro, K. F., Differential action of polycyclic aromatic hydrocarbons on endogenous estrogen-responsive genes and on a transfected estrogen-responsive reporter in MCF-7 cells. Toxicology and Applied Pharmacology, 196, 58–67 (2004).PubMedCrossRefGoogle Scholar
- Li, C., Takahashi, S., Taneda, S., Furuta, C., Watanabe, G., Suzuki, A. K., and Taya, K., Impairment of testicular function in adult male Japanese quail (Coturnix japonica) after a single administration of 3-methyl-4-nitrophenol in diesel exhaust particles. Journal of Endocrinology, 189, 555–564 (2006).PubMedCrossRefGoogle Scholar
- Machala, M., Ciganek, M., L., Bláha, L., Minksová, K., and Vondráèk, J., Aryl hydrocarbon receptor-mediated and estrogenic activities of oxygenated polycyclic aromatic hydrocarbons and azaarenes originally identified in extracts of river sediments. Environmental Toxicology and Chemistry, 20, 2736–2743 (2001).PubMedCrossRefGoogle Scholar
- Martin, M. B., Reiter, R., Pham, T., Avellanet, Y. R., Camara, J., Lahm, M., Pentecost, E., Pratap, K., Gilmore, B. A., Divekar, S., Dagata, R. S., Bull, J. L., and Stoica, A., Estrogen-like activity of metals in MCF-7 breast cancer cells. Endocrinology, 144, 2425–2436 (2003).PubMedCrossRefGoogle Scholar
- Navas, J. M. and Segner, H., Antiestrogenic activity of anthropogenic and natural chemicals. Environ. Sci. Pollut. Res., 5, 75–82 (1998).Google Scholar
- Perez, P., Pulgar, R., Olea-Serrano, F., Villalobos, M., Rivas, A., Metzler, M., Pedraza, V., and Olea, N., The estrogenicity of bisphenol A-related diphenylalkanes with various substituents at the central carbon and the hydroxyl group. Environ. Health Perspect., 106, 167–174 (1998).PubMedCrossRefGoogle Scholar
- Plíšková, M., Vondráèek, J., Vojtìšek, B., Kozubík, A., and Machala, M., Deregulation of cell proliferation by polycyclic aromatic hydrocarbons in human breast carcinoma MCF-7 cells reflects both genotoxic and nongenotoxic events. Toxicological Sciences, 83, 246–256 (2005).PubMedCrossRefGoogle Scholar
- Safe, S., Dietary and environmental estrogens and antiestrogens and their possible role in human disease. Environ. Sci. Pollut. Res., 1, 29–33 (1994).Google Scholar
- Taneda, S., Mori, Y., Kamata, K., Hayashi, H., Furuta, C., Li, C., Seki, K., Sakushima, A., Yoshino, S., Yamaki, K., Watanabe, G., Taya, K., and Suzuki, A. K., Estrogenic and anti-estrogenic activity of nitrophenols in diesel exhaust particles (DEP). Biol. Pharm. Bull., 27, 835–837 (2004).PubMedCrossRefGoogle Scholar
- Topinka, J., Schwarz, L. R., Kiefer, F., Wiebel, F. J., Gajdoš, O., Vidová, P., Dobiáš, L., Fried, M., Šrám, R.J., and Wolff, T., DNA adduct formation in mammalian cell cultures by polycyclic aromatic hydrocarbons (PAH) and nitro-PAH in coke oven emission extract. Mutation Research, 419, 91–105 (1998).PubMedGoogle Scholar
- U.S. Environmental Protection Agency (EPA), 2002. Health assessment document for diesel engine exhaust, Prepared by the National Center for Environmental Assessment, Washington, DC, for the Office of Transportation and Air Quality; EPA/600/8-90/057F.Google Scholar