METHODS: In this study, a cecal ligation and puncture (CLP)-induced sepsis model and DVT mouse model were constructed by inferior vena cava ligation. The levels of serum inflammatory factors and adhesion molecules were measured in each group, and the thrombus weight and size, hematoxylin-eosin staining, collagen fiber tissue, and transcriptome of the venous wall were analyzed. The activation of the Wnt/β-catenin signal was evaluated by quantitative real-time polymerase chain reaction, Western blotting, ELISA, and immunohistochemical and immunofluorescence methods.
RESULTS: Sepsis significantly promoted the formation of venous wall collagen fibers and DVT. In addition, Porcn significantly upregulated and activated the Wnt/β-catenin signaling pathway in sepsis mouse models with DVT. In contrast, the Wnt signaling inhibitor LGK974 was found to improve the survival rate, decrease thrombosis, and inhibit the expression of inflammation and adhesion molecules in sepsis mice with DVT. Therefore, activation of the Wnt/β-catenin signal may promote the formation of DVT in sepsis mice.
CONCLUSIONS: LGK974 protects against DVT formation in sepsis mice by inhibiting the activation of the Wnt/β-catenin signal and down-regulating the production of proinflammatory cytokines, PAI-1, and adhesion molecules. LGK974 may be a new candidate for the treatment of sepsis complicated with DVT.
方法:在本研究中,通过下腔静脉结扎建立盲肠结扎穿孔(CLP)诱导的脓毒症模型和DVT小鼠模型。检测各组血清炎症因子和黏附分子水平,血栓的重量和大小,苏木精-伊红染色,胶原纤维组织,并分析了静脉壁的转录组。通过定量实时聚合酶链反应评估Wnt/β-catenin信号的激活,西方印迹,ELISA,免疫组织化学和免疫荧光方法。
结果:脓毒症显著促进静脉壁胶原纤维和DVT的形成。此外,Porcn在DVT脓毒症小鼠模型中显著上调和激活Wnt/β-catenin信号通路。相比之下,发现Wnt信号抑制剂LGK974可以提高生存率,减少血栓形成,并抑制脓毒症DVT小鼠炎症和粘附分子的表达。因此,Wnt/β-catenin信号的激活可能促进脓毒症小鼠DVT的形成。
结论:LGK974通过抑制Wnt/β-catenin信号的激活和下调促炎细胞因子的产生来保护脓毒症小鼠DVT的形成,PAI-1和粘附分子。LGK974可能是治疗脓毒症并发DVT的新候选药物。