Estrogenic activity of procymidone in rainbow trout (Oncorhynchus mykiss) hepatocytes: a possible mechanism of action.
It is known that procymidone modifies sexual differentiation in vivo and in vitro, and that it induces vitellogenin (Vtg) synthesis in primary cultured rainbow trout hepatocytes. The aim of this study was to evaluate the mechanism underlying this latter in vitro estrogenic action. The cells were treated for 24 h with procymidone 150 microM (with 17beta-estradiol [E2] 20 microM as a positive control) combined with an estrogen receptor (ER) antagonist (tamoxifen 20 microM or ICI 182,780 1 microM) or, given the drug toxic action on the production of reactive oxygen species (ROS), a free radical scavenger (alpha-tocopherol 30 microM). The results from ELISA experiments provided evidence that procymidone Vtg-induction is inhibited by ER antagonists and by alpha-tocopherol suggesting that both ER and ROS are involved in this effect. The ROS detection revealed that the treatment with alpha-tocopherol and tamoxifen completely prevented ROS induction by procymidone, that was not inhibited by ICI 182,780. In exploring the mechanism mediating these events and its timing, we found that procymidone induced mitogen-activated protein kinase (MAPK) at 30 and 60 min, and that this effect was blocked by co-treatment with alpha-tocopherol. In summary, the results of the study clearly support the idea that the estrogenic activity of procymidone in primary cultured trout hepatocytes is mediated by ROS production, and that this activity is similar to that of the ligand-independent ER activation involving MAPK.
Radice
Sonia
S
Department of Pharmacology, Chemotherapy and Medical Toxicology "E. Trabucchi", University of Milan, Via Vanvitelli 32, 20129 Milan, Italy. sonia.radice@unimi.it
Fumagalli
Roberta
R
Chiesara
Enzo
E
Ferraris
Michela
M
Frigerio
Silvia
S
Marabini
Laura
L
D000818:Animals; D001643:Bridged Bicyclo Compounds / Q000633:toxicity*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004338:Drug Combinations; D004958:Estradiol / Q000031:analogs & derivatives* / Q000494:pharmacology; D004965:Estrogen Antagonists / Q000494:pharmacology; D004968:Estrogens, Non-Steroidal / Q000633:toxicity*; D005260:Female; D000077267:Fulvestrant; D005659:Fungicides, Industrial / Q000633:toxicity*; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D020929:Mitogen-Activated Protein Kinase Kinases / Q000096:biosynthesis; D017686:Oncorhynchus mykiss; D017382:Reactive Oxygen Species / Q000378:metabolism; D013629:Tamoxifen / Q000494:pharmacology; D014819:Vitellogenins / Q000378:metabolism; D024502:alpha-Tocopherol / Q000494:pharmacology
D001643:Bridged Bicyclo Compounds; D004338:Drug Combinations; D004965:Estrogen Antagonists; D004968:Estrogens, Non-Steroidal; D005659:Fungicides, Industrial; D017382:Reactive Oxygen Species; D014819:Vitellogenins; D013629:Tamoxifen; D000077267:Fulvestrant; D004958:Estradiol; D020929:Mitogen-Activated Protein Kinase Kinases; C035988:procymidone; D024502:alpha-Tocopherol