■脑微血管内皮细胞(BMEC)功能丧失是缺血性卒中发展的关键事件,可能受到环状RNA(circRNAs)失调的影响。我们的目的是揭示circRNAFKBP脯氨酸异构酶3(circFKBP3)在缺血性卒中细胞模型中的作用。
■在氧葡萄糖剥夺(OGD)治疗的人BEMC(HBMECs)中构建缺血性中风的细胞模型。进行定量实时PCR(qPCR)和蛋白质印迹以分析circFKBP3,miR-766-3p和TNF受体相关因子3(TRAF3)的表达。CCK-8Transwell,伤口愈合,流式细胞术,实施试管形成和ELISA测定以监测细胞活力,迁移,凋亡,血管生成和炎症产生。通过双荧光素酶验证了miR-766-3p与circFKBP3或TRAF3之间的假定结合关系,RIP和下拉测定。
■CircFKBP3表达在OGD处理的HBMECs中升高。OGD抑制HBMEC活力,迁移,血管生成,并引起细胞凋亡和炎症的产生,而circFKBP3的敲除减弱了这些作用。CircFKBP3与miR-766-3p相互作用,和circFKBP3缺失抑制的HBMEC功能丧失和炎症通过miR-766-3p抑制恢复。CircFKBP3靶向miR-766-3p调控TRAF3表达。通过TRAF3过表达恢复MiR-766-3p富集抑制的HBMEC功能丧失和炎症。
■CircFKBP3缺失通过控制miR-766-3p/TRAF3轴减轻了OGD诱导的HBMECs功能丧失和炎症反应。
UNASSIGNED: Brain microvascular endothelial cell (BMEC) functions loss is a key event in the development of ischemic stroke, which may be affected by the dysregulation of circular RNAs (circRNAs). We aimed to unveil the role of circRNA FKBP Prolyl Isomerase 3 (circFKBP3) in cell models of ischemic stroke.
UNASSIGNED: Cell models of ischemic stroke were constructed in human BEMCs (HBMECs) with the treatment of oxygen glucose deprivation (
OGD). Quantitative real-time PCR (qPCR) and western blotting were conducted for expression analysis of circFKBP3, miR-766-3p and TNF receptor associated factor 3 (TRAF3). CCK-8, transwell, wound healing, flow cytometry, tube formation and ELISA assays were implemented to monitor cell viability, migration, apoptosis, angiogenesis and inflammation production. The putative binding relationship between miR-766-3p and circFKBP3 or TRAF3 was validated by dual-luciferase, RIP and pull-down assays.
UNASSIGNED: CircFKBP3 expression was elevated in
OGD-treated HBMECs.
OGD suppressed HBMEC viability, migration, angiogenesis, and provoked cell apoptosis and inflammation production, while knockdown of circFKBP3 attenuated these effects. CircFKBP3 interacted with miR-766-3p, and circFKBP3 absence-repressed HBMEC function loss and inflammation were recovered by miR-766-3p inhibition. CircFKBP3 targeted miR-766-3p to regulate TRAF3 expression. MiR-766-3p enrichment-repressed HBMEC function loss and inflammation were recovered by TRAF3 overexpression.
UNASSIGNED: CircFKBP3 absence alleviated
OGD-induced function loss and inflammatory responses of HBMECs via governing the miR-766-3p/TRAF3 axis.
CircFKBP3 expression is elevated in
OGD-treated HBMECs.OGD-induced HBMEC function loss and inflammation are alleviated by circFKBP3 absence.CircFKBP3 directly targets miR-766-3p to regulate TRAF3 expression.