Raheem Kabir A, Marei Waleed F A, Campbell Bruce K, Fouladi-Nashta Ali A
Reproduction Group, Department of Comparative Biomedical Sciences, Royal Veterinary College, North Mymms, Hertfordshire, UK.
Division of Obstetrics and Gynaecology, Queens Medical Centre, School of Medicine, University of Nottingham, Nottingham, UK.
Theriogenology. 2016 Jun;85(9):1635-1643. doi: 10.1016/j.theriogenology.2016.01.018. Epub 2016 Jan 30.
In this study, we investigated the expression of mucin 1 (MUC1) mRNA and protein in sheep endometrium at different time points during follicular and luteal phases of estrous cycle, and also determined the effect of steroid hormone treatments and interferon tau (IFNτ) on MUC1 mRNA expression in endometrial cell culture in vitro. In experiment one, 15 Welsh mountain ewes were synchronized to a common estrus and killed at precise stages of estrous cycle corresponding to (1) pre-LH peak, (2) LH peak, (3) post-LH peak, (4) early luteal, and (5) mid-luteal. Reproductive tracts were harvested and mRNA was extracted from the endometrial tissues. Parts of the uterine horns were fixed for immunohistochemistry. In experiment two, mixed populations of ovine endometrial cells (from slaughterhouse material collected at the postovulatory stage of the estrous cycle) were cultured to 70% confluence before treatment with (1) progesterone (P4, 10 ng/mL, for 48 hours), (2) estradiol (E2, 100 pg/mL, for 48 hours), or with (3) E2 priming for 12 hours (100 pg/mL) followed by P4 (10 ng/mL) for 36 hours. These were compared with: (4) IFNτ (10 ng/mL, for 48 hours), and (5) basic medium (Dulbecco Modified Eagle Medium /F12) as control. The results showed that MUC1 mRNA and protein expression in sheep endometrium were highest during the midluteal stage and very low during the post-LH period compared with the other stages (P < 0.05). MUC1 immunostaining in the luminal epithelium was apically restricted and was not significantly different across all stages of estrous cycle except at the post-LH peak where it was significantly low. In cell culture, MUC1 mRNA expression was significantly upregulated by both steroids either singly or in combination (P < 0.05), and downregulated in the presence of IFNτ. In conclusion, endometrial MUC1 expression is cyclically regulated by both E2 and P4in vivo and in vitro, and directly downregulated by IFNτ treatment in vitro.
在本研究中，我们调查了发情周期卵泡期和黄体期不同时间点绵羊子宫内膜中粘蛋白1（MUC1）mRNA和蛋白的表达情况，并确定了类固醇激素处理和干扰素τ（IFNτ）对体外培养的子宫内膜细胞中MUC1 mRNA表达的影响。在实验一中，15只威尔士山地母羊被同步至共同发情期，并在发情周期的精确阶段处死，这些阶段分别对应于：（1）促黄体生成素（LH）峰前，（2）LH峰，（3）LH峰后，（4）黄体早期，以及（5）黄体中期。采集生殖道，从子宫内膜组织中提取mRNA。部分子宫角固定用于免疫组织化学分析。在实验二中，将绵羊子宫内膜细胞混合群体（取自发情周期排卵后期收集的屠宰场材料）培养至70%汇合度，然后分别用以下物质处理：（1）孕酮（P4，10 ng/mL，处理48小时），（2）雌二醇（E2，100 pg/mL，处理48小时），或（3）先用E2预处理12小时（100 pg/mL），然后用P4（10 ng/mL）处理36小时。将这些处理组与以下组进行比较：（4）IFNτ（10 ng/mL，处理48小时），以及（5）基础培养基（杜氏改良 Eagle 培养基/F12）作为对照。结果表明，与其他阶段相比，绵羊子宫内膜中MUC1 mRNA和蛋白表达在黄体中期最高，在LH峰后时期非常低（P < 0.05）。管腔上皮中的MUC1免疫染色位于顶端，除LH峰后显著降低外，在发情周期的所有阶段无显著差异。在细胞培养中，单独或联合使用两种类固醇均显著上调MUC1 mRNA表达（P < 0.05），而在IFNτ存在时则下调。总之，子宫内膜MUC1表达在体内和体外均受E2和P4的周期性调节，且在体外受IFNτ处理直接下调。
