Institute of Liver Diseases, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China.
Liuhe District Hospital of Traditional Chinese Medicine, Nanjing, Jiangsu Province, China.
BMC Gastroenterol. 2024 May 13;24(1):161. doi: 10.1186/s12876-024-03253-4.
Portal vein thrombosis (PVT) is a common complication of liver cirrhosis that can aggravate portal hypertension. However, there are features of both PVT and cirrhosis that are not recapitulated in most current animal models. In this study, we aimed to establish a stable animal model of PVT and cirrhosis, intervene with anticoagulant, and explore the related mechanism.
First, 49 male SD rats received partial portal vein ligation (PPVL), and 44 survival rats were divided into 6 groups: PPVL control group; 4-week, 6 -week, 8-week, and 10-week model group; and the rivaroxaban (RIVA)-treated group. The rats were intoxicated with or without carbon tetrachloride (CCl) for 4-10 weeks. Seven normal rats were used as the normal controls. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels and parameters for blood coagulation were all assayed with kits. Liver inflammation, collagen deposition and hydroxyproline (Hyp) levels were also measured. The extrahepatic macro-PVT was observed via portal vein HE staining, etc. The intrahepatic microthrombi was stained via fibrin immunohistochemistry. The portal blood flow velocity (PBFV) and diameter were detected via color Doppler ultrasound. Vascular endothelial injury was evaluated by von Willebrand Factor (vWF) immunofluorescence. Fibrinolytic activity was estimated by western blot analysis of fibrin and plasminogen activator inhibitor-1 (PAI-1).
After PPVL surgery and 10 weeks of CCl intoxication, a rat model that exhibited characteristics of both cirrhosis and extra and intrahepatic thrombi was established. In cirrhotic rats with PVT, the PBFV decreased, both factors of pro- and anti-coagulation decreased, but with relative hypercoagulable state, vascular endothelial injured, and fibrinolytic activity decreased. RIVA-treated rats had improved coagulation function, increased PBFV and attenuated thrombi. This effect was related to the improvements in endothelial injury and fibrinolytic activity.
A new rat model of PVT with cirrhosis was established through partial portal vein ligation plus CCl intoxication, with the characteristics of macrothrombi at portal veins and microthrombi in hepatic sinusoids, as well as liver cirrhosis. Rivaroxaban could attenuate PVT in cirrhosis in the model rats. The underlying mechanisms of PVT formation in the rat model and pharmacological action of rivaroxaban are related to the regulation of portal blood flow, coagulant factors, and vascular endothelial cell function.
门静脉血栓形成（PVT）是肝硬化的常见并发症，可加重门静脉高压。然而，大多数当前的动物模型都没有再现 PVT 和肝硬化的特征。本研究旨在建立一种稳定的 PVT 和肝硬化动物模型，进行抗凝干预，并探讨相关机制。
首先，49 只雄性 SD 大鼠接受部分门静脉结扎（PPVL），44 只存活大鼠分为 6 组：PPVL 对照组；4 周、6 周、8 周和 10 周模型组；以及利伐沙班（RIVA）治疗组。大鼠用或不用四氯化碳（CCl）处理 4-10 周。7 只正常大鼠作为正常对照组。使用试剂盒检测血清丙氨酸氨基转移酶（ALT）和天冬氨酸氨基转移酶（AST）水平以及凝血参数。还测量了肝炎症、胶原沉积和羟脯氨酸（Hyp）水平。通过门静脉 HE 染色等观察肝外大 PVT，通过纤维蛋白免疫组化观察肝内微血栓。通过彩色多普勒超声检测门静脉血流速度（PBFV）和直径。通过血管性血友病因子（vWF）免疫荧光评估血管内皮损伤。通过纤维蛋白和纤溶酶原激活物抑制剂-1（PAI-1）的 Western blot 分析评估纤溶活性。
在 PPVL 手术后和 CCl 中毒 10 周后，建立了一种同时具有肝硬化和肝内外血栓特征的大鼠模型。在患有 PVT 的肝硬化大鼠中，PBFV 降低，促凝和抗凝因子均降低，但存在相对高凝状态、血管内皮损伤和纤溶活性降低。RIVA 治疗的大鼠凝血功能改善，PBFV 增加，血栓减轻。这种作用与内皮损伤和纤溶活性的改善有关。
通过部分门静脉结扎加 CCl 中毒建立了一种具有肝硬化的新型大鼠 PVT 模型，其特征为门静脉大血栓和肝窦内微血栓，以及肝硬化。利伐沙班可减轻模型大鼠肝硬化中的 PVT。大鼠模型中 PVT 形成的潜在机制和利伐沙班的药理作用与门静脉血流、凝血因子和血管内皮细胞功能的调节有关。
