Identification of estrogenic and antiestrogenic activities of respirable diesel exhaust particles by bioassay-directed fractionation.
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 partitioning (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.
Oh
Seung Min
SM
College of Pharmacy, Sungkyunkwan University, Gangan-Gu, Suwon 440-746, Korea.
Ryu
Byung Taek
BT
Chung
Kyu Hyuck
KH
D001681:Biological Assay; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003627:Data Interpretation, Statistical; D052244:Endocrine Disruptors / Q000633:toxicity*; D004965:Estrogen Antagonists / Q000633:toxicity*; D004968:Estrogens, Non-Steroidal / Q000633:toxicity*; D005260:Female; D006801:Humans; D008752:Methylene Chloride; D052638:Particulate Matter / Q000633:toxicity; D012997:Solvents; D001335:Vehicle Emissions / Q000633:toxicity*
D052244:Endocrine Disruptors; D004965:Estrogen Antagonists; D004968:Estrogens, Non-Steroidal; D052638:Particulate Matter; D012997:Solvents; D001335:Vehicle Emissions; D008752:Methylene Chloride