Department of Gastroenterology, General Hospital of Northern Theater Command, Shenyang, China; Department of Pharmacy, General Hospital of Northern Theater Command, Shenyang, China; Department of Clinical Pharmacy, Shenyang Pharmaceutical University, Shenyang, China.
Department of Gastroenterology, General Hospital of Northern Theater Command, Shenyang, China.
Thromb Res. 2024 Jun;238:208-221. doi: 10.1016/j.thromres.2024.04.030. Epub 2024 May 1.
BACKGROUND & AIMS: Nonselective β blockers (NSBBs) facilitate the development of portal vein thrombosis (PVT) in liver cirrhosis. Considering the potential effect of NSBBs on neutrophils and neutrophil extracellular traps (NETs), we speculated that NSBBs might promote the development of PVT by stimulating neutrophils to release NETs.
Serum NETs biomarkers were measured, use of NSBBs was recorded, and PVT was evaluated in cirrhotic patients. Carbon tetrachloride and ferric chloride (FeCl) were used to induce liver fibrosis and PVT in mice, respectively. After treatment with propranolol and DNase I, neutrophils in peripheral blood, colocalization and expression of NETs in PVT specimens, and NETs biomarkers in serum were measured. Ex vivo clots lysis analysis was performed and portal vein velocity and coagulation parameters were tested.
Serum MPO-DNA level was significantly higher in cirrhotic patients treated with NSBBs, and serum H3Cit and MPO-DNA levels were significantly higher in those with PVT. In fibrotic mice, following treatment with propranolol, DNase I significantly shortened the time of FeCl-induced PVT formation, lowered the peripheral blood neutrophils labelled by CD11b/Ly6G, inhibited the positive staining of H3Cit and the expression of H3Cit and MPO proteins in PVT tissues, and reduced serum nucleosome level. Furthermore, the addition of DNase I to tissue plasminogen activator (tPA) significantly accelerated clots lysis as compared with tPA alone. Propranolol reduced portal vein velocity in fibrotic mice, but did not influence coagulation parameters.
Our study provides a clue to the potential impact of NETs formation on the association of NSBBs with the development of PVT.
非选择性β受体阻滞剂（NSBBs）可促进肝硬化患者门静脉血栓形成（PVT）。鉴于 NSBBs 对中性粒细胞和中性粒细胞胞外诱捕网（NETs）的潜在影响，我们推测 NSBBs 可能通过刺激中性粒细胞释放 NETs 而促进 PVT 的发展。
检测肝硬化患者的血清 NETs 生物标志物，记录 NSBBs 的使用情况，并评估 PVT。分别用四氯化碳和三氯化铁（FeCl）诱导小鼠肝纤维化和 PVT。用普萘洛尔和 DNA 酶 I 处理后，检测外周血中性粒细胞、PVT 标本中 NETs 的共定位和表达，以及血清中的 NETs 生物标志物。进行体外血栓溶解分析，并检测门静脉速度和凝血参数。
接受 NSBBs 治疗的肝硬化患者血清髓过氧化物酶-DNA 水平明显升高，PVT 患者血清 H3Cit 和 MPO-DNA 水平明显升高。在纤维化小鼠中，用普萘洛尔处理后，DNA 酶 I 显著缩短了 FeCl 诱导的 PVT 形成时间，降低了外周血中 CD11b/Ly6G 标记的中性粒细胞，抑制了 PVT 组织中 H3Cit 和 H3Cit 和 MPO 蛋白的阳性染色，并降低了血清核小体水平。此外，与单独使用组织型纤溶酶原激活剂（tPA）相比，向 tPA 中添加 DNA 酶 I 可显著加速血栓溶解。普萘洛尔降低了纤维化小鼠的门静脉速度，但不影响凝血参数。
我们的研究为 NETs 形成对 NSBBs 与 PVT 发展之间的关联的潜在影响提供了线索。
