Department of Obstetrics and Gynecology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Department of Obstetrics and Gynecology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Placenta. 2021 Mar;106:67-78. doi: 10.1016/j.placenta.2021.02.014. Epub 2021 Feb 26.
Preeclampsia is characterized by overactive inflammation at the uteroplacental interface, leading to trophoblasts dysfunction. 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase 3 (PFKFB3) is a crucial glycolytic regulator which has recently been found to participate in the pathological inflammatory states. This study aimed to investigate the role of PFKFB3 in the inflammation-induced damage in trophoblasts, and elucidate the underlying mechanisms.
Immunohistochemistry, qRT-PCR, and Western blot analysis (WB) were used to detect the expression of PFKFB3 in preeclamptic and normal placentas. Lipopolysaccharide (LPS)-induced HTR8/SVneo cells were established as the in vitro model to simulate the overactive inflammation at the uteroplacental interface of PE, which were subsequently transfected with PFKFB3 siRNA. The expression of PFKFB3, NF-κB-p-p65, phosphorylation states of NF-κB-p65, ICAM-1, Bcl-2, BAX, and MMP2 were detected by WB. qRT-PCR was used to detect the expression of TNF-α and IL-1β. The ICAM-1 expression was also reflected by monocyte adhesion assay. Reactive Oxygen Species (ROS) levels were detected by DCFH-DA (2,7-Dichlorodi-hydrofluorescein diacetate). Apoptosis was detected using Annexin V-FITC staining. Migration and invasion were measured by wound-healing and transwell assays.
PFKFB3 was up-regulated in the preeclamptic placenta. In LPS-treated HTR-8/Svneo cells, the inhibition of PFKFB3 blocked the NF-κB signal pathway, thereby downregulating the expression of proinflammatory cytokines and adhesion molecules, meanwhile, PFKFB3 knockdown significantly alleviated monocyte adhesion, oxidative stress, apoptosis, and reinstated migration and invasive capacity.
PFKFB3 controls the LPS-induced inflammation via the NF-κB pathway and impacts trophoblasts function such as adhesion, oxidative stress, apoptosis, migration, and invasion, thereby potentially participating in the preeclamptic etiopathogenesis.
子痫前期的特征是胎盘界面过度活跃的炎症反应，导致滋养层细胞功能障碍。6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶 3（PFKFB3）是一种关键的糖酵解调节因子，最近发现它参与了病理性炎症状态。本研究旨在探讨 PFKFB3 在滋养层细胞炎症诱导损伤中的作用，并阐明其潜在机制。
免疫组织化学、qRT-PCR 和 Western blot 分析（WB）用于检测子痫前期和正常胎盘组织中 PFKFB3 的表达。脂多糖（LPS）诱导的 HTR8/SVneo 细胞被建立为体外模型，以模拟 PE 胎盘界面的过度活跃炎症，随后用 PFKFB3 siRNA 转染。通过 WB 检测 PFKFB3、NF-κB-p-p65、NF-κB-p65 磷酸化状态、ICAM-1、Bcl-2、BAX 和 MMP2 的表达。qRT-PCR 用于检测 TNF-α 和 IL-1β 的表达。单核细胞黏附实验也反映了 ICAM-1 的表达。通过 DCFH-DA（2,7-二氯二氢荧光素二乙酸酯）检测活性氧（ROS）水平。使用 Annexin V-FITC 染色检测细胞凋亡。通过划痕愈合和 Transwell 测定法测量迁移和侵袭。
子痫前期胎盘组织中 PFKFB3 上调。在 LPS 处理的 HTR-8/Svneo 细胞中，PFKFB3 的抑制阻断了 NF-κB 信号通路，从而下调了促炎细胞因子和黏附分子的表达，同时，PFKFB3 敲低显著减轻了单核细胞黏附、氧化应激、细胞凋亡，并恢复了迁移和侵袭能力。
PFKFB3 通过 NF-κB 途径控制 LPS 诱导的炎症，并影响滋养层细胞的功能，如黏附、氧化应激、细胞凋亡、迁移和侵袭，从而可能参与子痫前期的发病机制。
