Department of Gastroenterology and Hepatology, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
Department of Ultrasound, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
Int J Mol Sci. 2024 Aug 22;25(16):9099. doi: 10.3390/ijms25169099.
Portal vein thrombosis (PVT) is a challenging and controversial complication of cirrhosis. Experimental models that reproduce cirrhotic PVT and effective pharmacological therapies are limited. We aimed to investigate the nature course and mechanisms of PVT in cirrhosis. A novel PVT model was developed via two-step total portal vein ligation in healthy and thioacetamide (TAA)-cirrhotic rats. Circulating and liver-infiltrating neutrophils were isolated from individuals with cirrhosis to examine neutrophil extracellular traps (NETs) and explore their unique characteristics and implications in PVT-associated fibrosis in cirrhosis. We further validated macrophage-myofibroblast transition (MMT) via multiplex immunofluorescence and single-cell sequencing. In the experimental model, cirrhosis promoted PVT development and portal vein intimal thickening. Interestingly, cirrhosis promoted spontaneous resolution of PVT due to instability of thrombus structure, along with pulmonary and intrahepatic clots. NETs-MMT mediate cirrhotic PVT and PVT-associated fibrosis, including fibrotic thrombus remodeling and increased hepatic collagen deposition. Mechanistically, caspase-4-dependent activation of neutrophils and GSDMD mediated the formation of NETs. The extracellular DNA of NETs promoted TGF-β1/Smad3-driven MMT. Inhibiting GSDMD with disulfiram suppressed cirrhotic PVT and prevented associated fibrosis. The cirrhotic PVT model reflected the following three main characteristics of cirrhotic PVT: spontaneous resolution, immunothrombosis, and intimal fibrosis. Targeting NETs with GSDMD inhibitors may serve as a new therapeutic concept to treat cirrhotic PVT.
门静脉血栓形成（PVT）是肝硬化的一种具有挑战性和争议性的并发症。能够复制肝硬化 PVT 并具有有效药理治疗作用的实验模型十分有限。我们旨在研究肝硬化中 PVT 的自然病程和机制。通过两步法结扎健康和硫代乙酰胺（TAA）诱导的肝硬化大鼠的总门静脉，建立了一种新的 PVT 模型。从肝硬化患者中分离循环和肝浸润中性粒细胞，以检查中性粒细胞胞外陷阱（NETs），并探索其在肝硬化相关 PVT 纤维化中的独特特征和意义。我们进一步通过多重免疫荧光和单细胞测序来验证巨噬细胞-肌成纤维细胞转化（MMT）。在实验模型中，肝硬化促进了 PVT 的发展和门静脉内膜增厚。有趣的是，由于血栓结构不稳定，以及肺内和肝内血栓，肝硬化促进了 PVT 的自发性消退。NETs-MMT 介导了肝硬化 PVT 和 PVT 相关纤维化，包括纤维性血栓重塑和肝胶原沉积增加。在机制上，中性粒细胞中 caspase-4 依赖性激活和 GSDMD 介导了 NETs 的形成。NETs 的细胞外 DNA 促进了 TGF-β1/Smad3 驱动的 MMT。用二硫苏糖醇抑制 GSDMD 可抑制肝硬化 PVT 并防止相关纤维化。肝硬化 PVT 模型反映了肝硬化 PVT 的三个主要特征：自发性消退、免疫性血栓形成和内膜纤维化。用 GSDMD 抑制剂靶向 NETs 可能成为治疗肝硬化 PVT 的一种新的治疗概念。
