雌激素对大鼠附睾上皮细胞非基因组作用及其机制研究

STUDY OF ESTROGEN NON-GENIC-GROUP ACTION AND ITS MECHANISM ON RAT PUBERTAL EPIDIDYMAL CELL

目的:研究雌激素对大鼠附睾上皮细胞非基因组作用及其机制。方法:青春期大鼠附睾上皮细胞,FLUO-3/AM标记,激光共聚焦显微镜技术观察。结果:17β-雌二醇能快速诱导附睾上皮细胞内Ca^(2+)[Ca^(2+)]_i升高,并具有剂量依赖性(10^(-2)～10^(-9)M);无Ca^(2+)液及2mM EGTA孵育附睾上皮细胞不影响雌二醇诱导效应;Ca^(2+)通道阻滞剂nifedipine、verapamil也不影响雌二醇诱导效应;而细胞内Ca^(2+)储器释放抑制剂neomycin能完全抑制雌二醇诱导效应。雌激素核内受体阻滞剂RU-486、mRNA转录抑制剂放线菌素D、蛋白质合成抑制剂放线菌酮等对雌二醇的效应均无作用。结论:雌激素以非基因组方式调控青春期大鼠附睾上皮细胞功能。

Objective: To study estrogen non-genic-group action and its mechanism on rat epididymal cell. Methods: The effects of estrogen on pubertal rat epididymal cell was investigated by laser-confocal-microscope. The rat epididymal intracellular calcium[Ca2+] was measured in cells loaded with the FLUO-3/AM. Results: There was an abrupt increase after the addition of estradiol-17. Estradiol-17β was effective between 10-2-10-9 M. Its effect was dose-dependent. The estra-diol-17-triggered [Ca2+]i surge was not affected by incubating the cells in Ca2+-free medium containing 2 mM EGTA or by pretreating them with Ca2+ channel blockers nifedipine or verapamil, but was abolished by pretreating them with neomycin. Its effect was not affected by pretreating the cells with the estrogen nuclear receptor antagonist RU-486, or the RNA and protein synthesis inhibitors actinomycin D and cycloheximide. Conclusion; These results suggest that the control of estrogen on pubertal rat epididymal cell function have been non-genic-group fashion.