小鼠药理性月经模型中孕酮撤退激活NF-κB状态研究

Pharmacologic withdrawal of progesterone activated NF-κB in the uterus of mouse menstruation model

目的:研究在小鼠药理性月经样模型中,阻断孕酮作用后子宫内膜组织崩解过程中核转录因子NF-κB激活状态。方法:切除卵巢后小鼠在诱导蜕膜化之后给予米非司酮处理模拟月经样出血,取部分子宫组织采用免疫组织化学、蛋白质印迹方法检测米非司酮处理(即孕酮撤退)后不同时期NF-κB激活及NF-κB上游信号分子的表达。结果:免疫组化结果显示,阻断孕酮作用后NF-κBp65显著入核,p50入核不明显。蛋白印迹实验显示,p50和p65蛋白发生了核浆转移,在孕酮撤退12h后核蛋白中表达量最高。NF-κB诱导性激酶、磷酸化IKK-β、磷酸化IκB-α在阻断孕酮作用后的各个时期都有表达,且各时相表达均有差异。结论:孕酮撤退可能通过NF-κB诱导性激酶、磷酸化IKK-β,继而磷酸化IκB-α通路激活NF-κB。

Objective： To investigate whether progesterone withdrawal activate NF - κB signaling in the uterus of mouse menstruation model. Methods： The menstruation model was established by administration of mifepristone （ RU486 ） after the induced decidualization of hormones - primed ovariectomized mice. The activation of NF - κB was evaluated by immunohistochemistry and western blot. Meanwhile, western blot were performed to determine the levels of the NF - κB inducing kinase （NIK）, IkB kinase 13 （IKK - β） and inhibitor of κB -α （IκB -α） that are all intermediates in the NF - κB signaling pathway. Results ： Immunohistochemically, NF - κB p65 was distinctly translocated from the cytoplasm to the nucleus in the uterine stromal and glandular epithelial cells of the mouse model after RU486 administration, whereas less p50 did so after administration of RU486. With western blotting, we observed that RU486 induced a remarkable increase in the expression of nuclear p65 and a gradual increase in p50, both of which peaked at 12 h of RU486 administration, which showed that NF -κB translocated from the cytoplasm to the nucleus. NIK, IKK - β and IκB -α differently expressed at all the checked time - points after RU486 treatment. Conclusion ： These results suggested that activation of NF - κB may be related to the function of two kinases （ NIK and IKK - β） after RU486 administration in mouse menstruation - like model.