Department of Reproductive Medical Center, Renmin Hospital of Wuhan University and Hubei Clinic Research Center for Assisted Reproductive Technology and Embryonic Development, Wuhan, Hubei 430060, China.
Department of Urology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, China.
Biol Reprod. 2022 Sep 12;107(3):834-845. doi: 10.1093/biolre/ioac100.
In the early stage of pregnancy, hypoxia in the placenta is of great significance to the migration and invasion of trophoblasts. In addition, changes to the polarity and activity of macrophages can affect embryo implantation, trophoblast migration and invasion, and vascular remodeling by affecting cytokine secretion. However, the mechanism of the effects of hypoxic conditions in the placenta on trophoblasts remains unknown. We used gene knockdown on macrophages, and drug treatment on trophoblasts, and cultured them under hypoxic and normoxic conditions. The cells were then subjected to wound-healing assays, Transwell cell invasion experiments, quantitative real-time reverse transcription Polymerase Chain Reaction (PCR), western blotting, and immunofluorescence. The polarization of macrophages in each group, the migration and invasion ability of trophoblasts, and changes to the phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway were detected. Hypoxic conditions induce M2 polarization of macrophages. The conditioned medium from macrophages under hypoxic conditions increased the migration and invasion of trophoblasts and enhanced the levels of phosphorylated (p)-PI3K and p-AKT in trophoblasts. After C-C motif chemokine ligand 5 knockdown in macrophages, the ability of conditioned medium from macrophages cultured under hypoxic conditions to promote the migration and invasion of trophoblasts was weakened significantly. The use of PI3K/AKT signaling pathway agonists could reverse the attenuation effect caused by C-C motif chemokine ligand 5 knockdown.
在妊娠早期，胎盘缺氧对滋养细胞的迁移和侵袭具有重要意义。此外，通过影响细胞因子分泌，改变巨噬细胞的极性和活性，也会影响胚胎着床、滋养细胞迁移和侵袭以及血管重塑。然而，胎盘缺氧对滋养细胞的影响机制尚不清楚。本研究通过基因敲低巨噬细胞和药物处理滋养细胞，并在缺氧和常氧条件下培养它们，然后进行划痕愈合试验、Transwell 细胞侵袭实验、实时定量聚合酶链反应（PCR）、Western blot 和免疫荧光检测。检测各组巨噬细胞的极化、滋养细胞的迁移和侵袭能力以及磷脂酰肌醇-4,5-二磷酸 3-激酶（PI3K）/蛋白激酶 B（AKT）信号通路的变化。缺氧条件诱导巨噬细胞 M2 极化。缺氧条件下培养的巨噬细胞条件培养基增加了滋养细胞的迁移和侵袭，并增强了滋养细胞中磷酸化（p）-PI3K 和 p-AKT 的水平。在巨噬细胞中敲低 C-C 基序趋化因子配体 5 后，缺氧条件下培养的巨噬细胞条件培养基促进滋养细胞迁移和侵袭的能力明显减弱。PI3K/AKT 信号通路激动剂的使用可以逆转 C-C 基序趋化因子配体 5 敲低引起的衰减作用。
