肿瘤坏死因子在小鼠药理性孕酮撤退月经样模型中的定位和表达

摘要

Objective:To investigate the localization and expression of tumor necrosis factor-alfa(TNF-α)in mouse menstrual-like model created by pharmacologic progesterone withdrawal. Methods: The menstrual-like model was prepared by administration of mifepristone ( RU486 ) after decidualization of endometrium in ovariectomized mice. The location of TNF-α protein in uterus was detected by immunohistochemistry. The mRNA expression of TNF-α was analyzed by real-time PCR at different time within 24 hours after RU486 administration. Results:The immunohistochemical results showed that TNF-α immunopositive cells were present in the focal subluminal vascular endothelial cells 0-8 hours after administration of mifepristone. The staining appeared in leukocytes in the myometrium after 12-16 hours and the stroma in the basal zone and the decidualized zone,as well as in the extracellular matrix(ECM)across the border of dissociating tissue after 24-48 hours. In addition,TNF-α was also present in repaired stromal cells around glandular epithelium. RU486 induced a remarkable increase in expression of TNF-α mRNA with maximal level at 12 and 16 hours. Conclusion: The localization of TNF-α in the model displays a regional character. Progesterone withdrawal can induce expression of TNF-α, which may be involved in the process of tissue breakdown in the model.%目的 在药理性撤退小鼠月经样模型中,观察孕酮撤退后,肿瘤坏死因子(TNF-α)的定位和表达情况.方法 卵巢切除的小鼠在诱导蜕膜化之后给予米非司酮即通过拮抗孕酮受体的方法(药理性撤退),导致孕酮撤退,并用免疫组化的方法观察米非司酮处理的不同时相,TNF-α在小鼠子宫内膜的定位；用实时PCR的方法观察TNF-α表达情况.结果 免疫组化结果显示米非司酮处理后0～8 h,TNF-α主要存在于腔周围的血管内皮细胞；12～16 h,主要存在于内流的白细胞；24～48 h,主要存在于崩解组织边缘的间质细胞区域和修复好腺上皮细胞周围的间质细胞.实时PCR结果显示TNF-α mRNA在孕酮撤退后24 h内都有表达,并且TNF-a表达量逐渐增高,在12h、16h达到峰值. 结论 在此小鼠月经样模型中,米非司酮处理即孕酮撤退后TNF-α在此模型子宫中定位呈区域性分布,并诱导TNF-α表达升高,可能参与了子宫内膜的崩解.