Vitellogenin (Vtg) is the precursor of yolk protein. Its expression and secretion are estrogen-regulated and are crucial for oocyte maturation. An in vitro xenoestro- gen screening model was established by measuring V...Vitellogenin (Vtg) is the precursor of yolk protein. Its expression and secretion are estrogen-regulated and are crucial for oocyte maturation. An in vitro xenoestro- gen screening model was established by measuring Vtg in- duction in cultured primary hepatocytes from crucian carp. Vtg production was detected by biotin-avidin sandwich ELISA method while Vtg and cytochrome P4501A1 (CYP1A1) mRNA induction were measured by semi- quan- titative PCR-primer dropping technique. Vtg and Vtg mRNA were dose-dependently induced by diethylstilbestrol (DES, 0.2—200 ng/mL) in hepatocytes of crucian carp. Co-treatment of the DES-induced hepatocytes with either 2,3,7,8-TCDD (TCDD, 0.1—4 pg/mL) or benzo[a]pyrene (B[a]P, 5—1000 ng/mL) resulted in a reduction of Vtg pro- duction and an increment of CYP1A1 mRNA expression both in a dose dependent manner, indicating the anti-estro- genic effects of the compounds. However, at lower tested concentrations, TCDD (0.1, 0.2 pg/mL), B[a]P (5 ng/mL) seemed to have a potentiating effect on Vtg expression and secretion, although by their own these compounds had no observable estrogenic effect on Vtg induction. Tamoxifen (a selective estrogen receptor modulators, 1 nmol/L—1 μmol/L), and β-naphtho-flavone (β-NF, an aryl hydrocarbon receptor inducing compounds, 2.5—1000 ng/mL) also were employed to study the possible interactions in DES-induced Vtg ex- pression. In co-treatment of the DES-induced hepatocytes with β-NF or tamoxifen, the decrease in Vtg production did parallel induction of CYP1A1 for β-NF, but tamoxifen inhib- ited Vtg induction did not parallel induced CYP1A1 expres- sion in all test concentrations. On the contrary, it was found that in co-treatment of the TCDD-induced hepatocytes with DES, TCDD induced CYP1A1 mRNA production was inhib- ited by DES also. These results implicated a possible cross talk between estrogen receptor- and aryl hydrocarbon re- ceptor-mediated pathways in the hepatocytes.展开更多
文摘Vitellogenin (Vtg) is the precursor of yolk protein. Its expression and secretion are estrogen-regulated and are crucial for oocyte maturation. An in vitro xenoestro- gen screening model was established by measuring Vtg in- duction in cultured primary hepatocytes from crucian carp. Vtg production was detected by biotin-avidin sandwich ELISA method while Vtg and cytochrome P4501A1 (CYP1A1) mRNA induction were measured by semi- quan- titative PCR-primer dropping technique. Vtg and Vtg mRNA were dose-dependently induced by diethylstilbestrol (DES, 0.2—200 ng/mL) in hepatocytes of crucian carp. Co-treatment of the DES-induced hepatocytes with either 2,3,7,8-TCDD (TCDD, 0.1—4 pg/mL) or benzo[a]pyrene (B[a]P, 5—1000 ng/mL) resulted in a reduction of Vtg pro- duction and an increment of CYP1A1 mRNA expression both in a dose dependent manner, indicating the anti-estro- genic effects of the compounds. However, at lower tested concentrations, TCDD (0.1, 0.2 pg/mL), B[a]P (5 ng/mL) seemed to have a potentiating effect on Vtg expression and secretion, although by their own these compounds had no observable estrogenic effect on Vtg induction. Tamoxifen (a selective estrogen receptor modulators, 1 nmol/L—1 μmol/L), and β-naphtho-flavone (β-NF, an aryl hydrocarbon receptor inducing compounds, 2.5—1000 ng/mL) also were employed to study the possible interactions in DES-induced Vtg ex- pression. In co-treatment of the DES-induced hepatocytes with β-NF or tamoxifen, the decrease in Vtg production did parallel induction of CYP1A1 for β-NF, but tamoxifen inhib- ited Vtg induction did not parallel induced CYP1A1 expres- sion in all test concentrations. On the contrary, it was found that in co-treatment of the TCDD-induced hepatocytes with DES, TCDD induced CYP1A1 mRNA production was inhib- ited by DES also. These results implicated a possible cross talk between estrogen receptor- and aryl hydrocarbon re- ceptor-mediated pathways in the hepatocytes.
