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铝对虹鳟(Oncorhynchus mykiss)原代肝细胞中17β-雌二醇诱导卵黄蛋白原生成的抑制作用。

Inhibitory effects of aluminium on vitellogenin induction by estradiol-17 beta in the primary culture of hepatocytes in the rainbow trout Oncorhynchus mykiss.

作者信息

Mugiya Y, Tanahashi A

机构信息

Laboratory of Physiology, Faculty of Fisheries, Hokkaido University, Hakodate, Japan.

出版信息

Gen Comp Endocrinol. 1998 Jan;109(1):37-43. doi: 10.1006/gcen.1997.6996.

Abstract

Effects of Al on estradiol-induced vitellogenin (VTG) induction were electrophoretically examined in primary hepatocyte cultures in rainbow trout. Hepatocytes were precultured for 2 days and then estradiol-17 beta (E2, 6 x 10(-6) M) and Al (10(-6)-10(-4) M) were added to the incubation medium. The hepatocytes were cultured for 5 more days. Spent media were analyzed by SDS-PAGE and the relative rate of VTG synthesis was evaluated by a measurement of the integrated optical density of the main VTG band and was expressed as the percentage of VTG to total proteins including the VTG. The addition of Al to the incubation medium had no effect on the viability of hepatocytes in the culture. However, it specifically reduced VTG synthesis by hepatocytes in a concentration-dependent way and there was a significant reduction at Al concentrations greater than 6 x 10(-5) M. VTG synthesis by E2-primed hepatocytes was also reduced by Al concentrations of more than 6 x 10(-5) M 2-6 days after Al addition. Enriched Ca concentrations (1.8 to 2.5 or 5.0 mM) in the incubation medium had no protective effect on the reduction of VTG synthesis by Al. These results suggest that the synthesis of VTG is more susceptible to Al than are other hepatocyte-derived proteins.

摘要

在虹鳟原代肝细胞培养物中,通过电泳检测了铝对雌二醇诱导的卵黄蛋白原(VTG)合成的影响。肝细胞预培养2天,然后向孵育培养基中添加17β-雌二醇(E2,6×10⁻⁶ M)和铝(10⁻⁶ - 10⁻⁴ M)。肝细胞再培养5天。对用过的培养基进行SDS-PAGE分析,并通过测量主要VTG条带的积分光密度来评估VTG合成的相对速率,以VTG占包括VTG在内的总蛋白的百分比表示。向孵育培养基中添加铝对培养的肝细胞活力没有影响。然而,它以浓度依赖的方式特异性地降低了肝细胞的VTG合成,在铝浓度大于6×10⁻⁵ M时显著降低。添加铝后2 - 6天,铝浓度大于6×10⁻⁵ M时,E2预处理的肝细胞的VTG合成也降低。孵育培养基中富集的钙浓度(1.8至2.5或5.0 mM)对铝降低VTG合成没有保护作用。这些结果表明,与其他肝细胞衍生蛋白相比,VTG的合成对铝更敏感。

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