Research Centre for Reproductive Health, School of Paediatrics and Reproductive Health, University of Adelaide, Adelaide, South Australia, Australia.
Biol Reprod. 2011 Jan;84(1):179-88. doi: 10.1095/biolreprod.110.085399. Epub 2010 Sep 23.
Macrophages accumulate within stromal tissue subjacent to the luminal epithelium in the mouse uterus during early pregnancy after seminal fluid exposure at coitus. To investigate their role in regulating epithelial cell expression of fucosylated structures required for embryo attachment and implantation, fucosyltransferase enzymes Fut1, Fut2 (Enzyme Commission number [EC] 2.4.1.69), and Fut4 (EC 2.4.1.214) and Muc1 and Muc4 mRNAs were quantified by quantitative real-time PCR in uterine epithelial cells after laser capture microdissection in situ or after epithelial cell coculture with macrophages or macrophage-secreted factors. When uterine macrophage recruitment was impaired by mating with seminal plasma-deficient males, epithelial cell Fut2 expression on Day 3.5 postcoitus (pc) was reduced compared to intact-mated controls. Epithelial cell Fut2 was upregulated in vitro by coculture with macrophages or macrophage-conditioned medium (MCM). Macrophage-derived cytokines LIF, IL1B, and IL12 replicated the effect of MCM on Fut2 mRNA expression, and MCM-stimulated expression was inhibited by anti-LIF and anti-IL1B neutralizing antibodies. The effects of acute macrophage depletion on fucosylated structures detected with lectins Ulex europaeus 1 (UEA-1) and Lotus tetragonolobus purpureas (LTP), or LewisX immunoreactivity, were quantified in vivo in Cd11b-dtr transgenic mice. Depletion of macrophages caused a 30% reduction in luminal epithelial UEA-1 staining and a 67% reduction in LewisX staining in uterine tissues of mice hormonally treated to mimic early pregnancy. Together, these data demonstrate that uterine epithelial Fut2 mRNA expression and terminal fucosylation of embryo attachment ligands is regulated in preparation for implantation by factors including LIF and IL1B secreted from macrophages recruited during the inflammatory response to insemination.
在交配后精液暴露的情况下，小鼠子宫在妊娠早期的腔上皮下的基质组织中，巨噬细胞会积累。为了研究它们在调节上皮细胞表达胚胎附着和植入所需的岩藻糖基化结构中的作用，通过激光捕获显微切割原位或上皮细胞与巨噬细胞或巨噬细胞分泌因子共培养后，定量实时 PCR 定量了子宫上皮细胞中的岩藻糖基转移酶 Fut1、Fut2（酶委员会编号 [EC]2.4.1.69）和 Fut4（EC2.4.1.214）以及 Muc1 和 Muc4 mRNA。当通过与缺乏精液的雄性交配来损害子宫巨噬细胞募集时，与完整交配对照组相比，交配后第 3.5 天（pc）的上皮细胞 Fut2 表达减少。体外与巨噬细胞或巨噬细胞条件培养基（MCM）共培养可上调上皮细胞 Fut2。巨噬细胞衍生的细胞因子 LIF、IL1B 和 IL12 复制了 MCM 对 Fut2 mRNA 表达的影响，而 MCM 刺激的表达被抗 LIF 和抗 IL1B 中和抗体抑制。在 Cd11b-dtr 转基因小鼠体内，通过凝集素 Ulex europaeus 1 (UEA-1) 和 Lotus tetragonolobus purpureas (LTP) 或 LewisX 免疫反应性检测到的急性巨噬细胞耗竭对岩藻糖基化结构的影响进行了量化。巨噬细胞耗竭导致激素处理以模拟早孕的小鼠子宫组织中腔上皮 UEA-1 染色减少 30%，LewisX 染色减少 67%。这些数据表明，包括 LIF 和 IL1B 在内的因子通过募集到与受精后炎症反应相关的巨噬细胞来调节子宫上皮 Fut2 mRNA 表达和胚胎附着配体的末端岩藻糖基化，为植入做准备。
