17β-雌二醇上调绵羊输卵管上皮细胞β-防御素-2(SBD-2)表达的可能信号通路研究

Up-regulation mechanism of 17β-estradiol onβ-Defensin-2(SBD-2)in ovine oviduct epithelial cells

【目的】探讨17β-雌二醇(E2)上调绵羊输卵管上皮细胞SBD-2表达的可能信号通路。【方法】分离培养绵羊输卵管上皮细胞,将第2代输卵管上皮细胞分为E2(10-8 mol/L)组、雌激素受体拮抗剂ICI182780(10-7mol/L)组、PKA阻断剂KT-5720(1μmol/L)组、PKC阻断剂H-7(50μmol/L)组、NF-κB阻断剂PDTC(50μmol/L)组及空白对照(Control)组,在不同阻断剂组加入各自阻断剂干预绵羊输卵管上皮细胞1h后,在各阻断剂组和E2组中再加入10-8 mol/L 17β-雌二醇培养6h,然后采用实时荧光定量RT-PCR方法检测各组SBD-2mRNA表达水平的变化。【结果】10-8 mol/L 17β-雌二醇可显著上调绵羊输卵管上皮细胞SBD-2mRNA的表达(P<0.05);雌激素受体拮抗剂ICI182780、NF-κΒ阻断剂PDTC、PKC阻断剂H-7均可阻断17β-雌二醇对SBD-2的上调作用,PKA阻断剂KT-5720对17β-雌二醇介导的SBD-2上调无明显影响。【结论】17β-雌二醇上调绵羊输卵管上皮细胞SBD-2的表达由雌激素核受体、NF-κΒ、PKC信号转导通路所介导,而PKA不参与17β-雌二醇对SBD-2的上调作用。

【Objective】This study aimed to investigate the mechanisms involving in the inhibitory effect of 17β-estradiol （E2） on β-Defensin-2 （SBD-2） in ovine oviduct epithelial cells.【Method】Ovine oviduct epithelial cells were isolated and cultured,and then the second generation cells were divided into 17β-estradiol （10-8 mol/L） group,estrogen nuclear receptor antagonists ICI182780 （10-7 mol/L） group,PKA antagonist KT-5720 （1 μmol/L） group,PKC antagonist H-7 （50 μmol/L） group,nuclear factor kappa B antagonist PDTC （50 μmol/L） group and control group （Control）.Different antagonists were added to each antagonist group to interfere ovine oviduct epithelial cells for 1 h,and then 17β-estradiol （10-8 mol/L） was added to each antagonist group and E2 group for 6 h.Real time PCR was used to detect the changes of SBD-2.【Result】The expression of SBD-2 mRNA was significantly up-regulated by 17β-estradiol （10-8 mol/L） （P〈0.05）.The effect was abrogated by ICI182780,PKC antagonist H-7 and NF-κΒ antagonist PDTC while PKA antagonist KT-5720 had no influence.【Conclusion】17β-estradiol induces SBD-2 mRNA expression in ovine oviduct epithelial cells,and the induction is mediated by signaling proteins of estrogen nuclear receptor,PKC and NF0κΒ rather than PKA.