Abstract：
OBJECTIVE: To explore the role of 14-3-3 protein and the Hippo and yes-associated protein 1 (YAP) signaling pathway in lipopolysaccharide (LPS)-induced vascular inflammation.
METHODS: Human umbilical vein endothelial cells (HUVECs) and C57B6 mice were treated with LPS to establish cell and animal models of vascular inflammation. Lentiviral transfection, Western blot, qPCR, immunofluorescence, immunohistochemistry, co-immunoprecipitation, and enzyme-linked immunosorbent assays were used to measure inflammatory factors and expression of 14-3-3 protein and phosphorylation of YAP at S127. HUVECs and C57B6 mice were pretreated with a YAP inhibitor, Verteporfin, to observe changes in YAP expression and downstream vascular inflammation.
RESULTS: LPS induced acute and chronic inflammatory responses in HUVECs and mice and upregulated the expression of several inflammatory factors. LPS also induced expression of 14-3-3 protein and phosphorylation of YAP at S127 in response to acute vascular inflammation and downregulated these markers in response to chronic vascular inflammation. Verteporfin reduced these LPS-induced effects on vascular inflammation.
CONCLUSIONS: In chronic vascular inflammation, 14-3-3 protein is downregulated, which promotes inflammation by increasing Hippo/YAP nuclear translocation.
METHODS: Human umbilical vein endothelial cells (HUVECs) and C57B6 mice were treated with LPS to establish cell and animal models of vascular inflammation. Lentiviral transfection, Western blot, qPCR, immunofluorescence, immunohistochemistry, co-immunoprecipitation, and enzyme-linked immunosorbent assays were used to measure inflammatory factors and expression of 14-3-3 protein and phosphorylation of YAP at S127. HUVECs and C57B6 mice were pretreated with a YAP inhibitor, Verteporfin, to observe changes in YAP expression and downstream vascular inflammation.
RESULTS: LPS induced acute and chronic inflammatory responses in HUVECs and mice and upregulated the expression of several inflammatory factors. LPS also induced expression of 14-3-3 protein and phosphorylation of YAP at S127 in response to acute vascular inflammation and downregulated these markers in response to chronic vascular inflammation. Verteporfin reduced these LPS-induced effects on vascular inflammation.
CONCLUSIONS: In chronic vascular inflammation, 14-3-3 protein is downregulated, which promotes inflammation by increasing Hippo/YAP nuclear translocation.
摘要：
目的：探讨14-3-3蛋白和Hippo与Yes相关蛋白1(YAP)信号通路在脂多糖(LPS)诱导的血管炎症中的作用。
方法：用LPS处理人脐静脉内皮细胞（HUVECs）和C57B6小鼠，建立血管炎症的细胞和动物模型。慢病毒转染，蛋白质印迹,qPCR,免疫荧光，免疫组织化学，免疫共沉淀,采用酶联免疫吸附试验检测S127时炎性因子、14-3-3蛋白表达和YAP磷酸化。HUVEC和C57B6小鼠用YAP抑制剂预处理，Verteporfin,观察YAP表达和下游血管炎症的变化。
结果：LPS可诱导HUVECs和小鼠的急性和慢性炎症反应，并上调多种炎症因子的表达。LPS还响应于急性血管炎症在S127诱导14-3-3蛋白的表达和YAP的磷酸化,并且响应于慢性血管炎症下调这些标志物。Verteporfin降低了这些LPS诱导的对血管炎症的影响。
结论：在慢性血管炎症中，14-3-3蛋白下调，通过增加Hippo/YAP核易位促进炎症。
方法：用LPS处理人脐静脉内皮细胞（HUVECs）和C57B6小鼠，建立血管炎症的细胞和动物模型。慢病毒转染，蛋白质印迹,qPCR,免疫荧光，免疫组织化学，免疫共沉淀,采用酶联免疫吸附试验检测S127时炎性因子、14-3-3蛋白表达和YAP磷酸化。HUVEC和C57B6小鼠用YAP抑制剂预处理，Verteporfin,观察YAP表达和下游血管炎症的变化。
结果：LPS可诱导HUVECs和小鼠的急性和慢性炎症反应，并上调多种炎症因子的表达。LPS还响应于急性血管炎症在S127诱导14-3-3蛋白的表达和YAP的磷酸化,并且响应于慢性血管炎症下调这些标志物。Verteporfin降低了这些LPS诱导的对血管炎症的影响。
结论：在慢性血管炎症中，14-3-3蛋白下调，通过增加Hippo/YAP核易位促进炎症。
