Department of Obstetrics and Gynecology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Oxid Med Cell Longev. 2022 Jun 14;2022:7202837. doi: 10.1155/2022/7202837. eCollection 2022.
Preeclampsia (PE) is a heterogeneous disease closely associated with the accelerated senescence of the placentas. Placental mesenchymal stem cells (PMSCs) modulate placental development, which is abnormally senescent in PE together with abnormal paracrine. Both pivotal in the placenta development, Toll-like receptor 4 (TLR4) and Hedgehog (HH) pathway are also tightly involved in regulating cellular senescence. This study was aimed at demonstrating that TLR4/HH pathway modulated senescence of placentas and PMSCs and . Preeclamptic and normal PMSCs were isolated. Smoothed agonist (SAG) and cyclopamine were used to activate and inhibit HH pathway, respectively. Lipopolysaccharide (LPS) was used to activate TLR4 and establish the classic PE-like rat model. qRT-PCR, Western blotting, and immunofluorescence were used to detect the expression of TLR4 and HH components (SHH, SMO, and Gli1). Cellular biological function such as proliferation, apoptosis, and migration was compared. Cell cycle analysis, -galactosidase staining, and the protein expressions of p16 and p53 were detected to analyze the cellular senescence. The secretion levels of human matrix metalloproteinase 9 (MMP-9) and soluble fms-like tyrosine kinase-1 (sFlt-1) were measured in the conditioned medium. Cell migration, invasion, and tube formation were analyzed in HTR8/SVneo cells or human umbilical vein endothelial cells (HUVECs). Our study demonstrated that activation of TLR4 accelerated senescence of PMSCs via suppressing HH pathway both and , accompanied by the detrimental paracrine to impair the uterine spiral artery remodeling and placental angiogenesis. Meanwhile, induction of HH pathway could alleviate PE-like manifestations, improve pregnancy outcomes, and ameliorate multiorgan injuries, suggesting that strengthening the HH pathway may serve as a potential therapy in PE.
子痫前期 (PE) 是一种异质性疾病，与胎盘的加速衰老密切相关。胎盘间充质干细胞 (PMSC) 调节胎盘发育，PE 中胎盘发育异常衰老，并伴有异常的旁分泌作用。Toll 样受体 4 (TLR4) 和 Hedgehog (HH) 通路在胎盘发育中都起着至关重要的作用，它们也紧密参与调节细胞衰老。本研究旨在证明 TLR4/ HH 通路调节胎盘和 PMSC 的衰老，以及。分离出子痫前期和正常的 PMSC。用平滑激动剂 (SAG) 和环巴胺分别激活和抑制 HH 通路。用脂多糖 (LPS) 激活 TLR4 并建立经典的 PE 样大鼠模型。用 qRT-PCR、Western blot 和免疫荧光检测 TLR4 和 HH 成分 (SHH、SMO 和 Gli1) 的表达。比较细胞生物学功能，如增殖、凋亡和迁移。通过细胞周期分析、β-半乳糖苷酶染色和 p16 和 p53 蛋白表达来分析细胞衰老。在条件培养基中测量人基质金属蛋白酶 9 (MMP-9) 和可溶性 fms 样酪氨酸激酶-1 (sFlt-1) 的分泌水平。在 HTR8/SVneo 细胞或人脐静脉内皮细胞 (HUVEC) 中分析细胞迁移、侵袭和管形成。我们的研究表明，TLR4 的激活通过抑制 HH 通路加速 PMSC 的衰老，同时伴随着有害的旁分泌作用，损害子宫螺旋动脉重塑和胎盘血管生成。同时，HH 通路的诱导可以减轻 PE 样表现，改善妊娠结局，改善多器官损伤，表明增强 HH 通路可能是 PE 的一种潜在治疗方法。
