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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 the non-genomic effect of estrogen on rat epididymal epithelial cells and its mechanism. Methods: Adolescent epididymal epithelial cells were stained with FLUO-3 / AM and observed with laser scanning confocal microscopy. Results: 17β-estradiol rapidly induced the increase of intracellular calcium Ca ~ (2 +) _ i in epididymal epithelial cells in a dose-dependent manner (10 ~ (-2) ~ 10 ~ (-9 ) M); Ca 2+ -free and 2 mM EGTA incubated epididymal epithelial cells did not affect the estradiol-induced effect; Ca 2+ channel blockers nifedipine and verapamil did not affect the estradiol-induced effect; The intracellular Ca ~ (2 +) release inhibitor neomycin completely inhibited the estradiol-induced effect. Estrogen nuclear receptor blocker RU-486, mRNA transcription inhibitor actinomycin D, protein synthesis inhibitor cycloheximide and so on have no effect on estradiol. Conclusion: Estrogen regulates epididymal epithelial cell function in adolescent rats by non-genomic methods.