Hyperuricemia enhances procoagulant activity of vascular endothelial cells through TMEM16F regulated phosphatidylserine exposure and microparticle release

Abstract

The link between serum uric acid (SUA) and the risk of venous thromboembolism (VTE) is well established. Recent data suggested a causative role of UA in endothelial cells (ECs) dysfunction. However, the molecular mechanism of high UA on thrombogenesis is unknown. We investigate whether high UA induce phosphatidylserine (PS) externalization and microparticle (MP) shedding in cultured EC, and contribute to UA‐induced hypercoagulable state. In the present study, we demonstrate that UA induces PS exposure and EMP release of EC in a concentration‐ and time‐dependent manner, which enhances the procoagulant activity (PCA) of EC and inhibited over 90% by lactadherin in vitro. Furthermore, hyperuricemic rat model was used to evaluate the development of thrombi following by flow stasis in the inferior vena cava (IVC). Hyperuricemia group is more likely to form large and hard thrombi compared with control. Importantly, we found that TMEM16F expression is significantly upregulated in UA‐treated EC, which is crucial for UA‐induced PS exposure and MP formation. Additionally, UA increases the generation of reactive oxygen species (ROS), lipid peroxidation, and cytosolic Ca2+ concentration in EC, which might contribute to increased TMEM16F expression. Using confocal microscopy, we also observed disruption of the actin cytoskeleton, suggesting that depolymerization of actin filaments might be required for TMEM16F activation and followed by PS exposure and membrane blebbing in UA‐treated EC. Our results demonstrate a thrombotic role of EC in hyperuricemia through TMEM16F‐mediated PS exposure and MPs release.

Shape-specific microfabricated particles for biomedical applications: a review

“…Moreover, the size is critical in determining the hemocompatibility of microparticles when administered parenterally. The human body naturally produces microparticles under pathologic conditions (e.g., cancer, endothelial alterations, inflammation) [59][60][61], resulting from membrane blebbing as part of cell apoptosis [62]. Artificial microparticles (e.g., DPNs) can mitigate this risk by minimizing protein adsorption (e.g., through their materials) and by tuning the particle materials properties (i.e., particle deformability and size).…”

Hyperuricemia: A key contributor to endothelial dysfunction in cardiovascular diseases

“…In addition, endothelial microparticles (MPs) participate in the mechanism underlying HUA‐induced coagulation. Yu et al 67 demonstrated that UA at high levels can induce cytoskeletal structure disruption, transmembrane protein 16F (TMEM16F) activation, phosphatidylserine (PS) externalization, and MP shedding in HUVECs by increasing the levels of intracellular calcium and ROS. After MP shedding and PS externalization on ECs, binding sites are made available for the coagulation factors FXA and the prothrombinase complex, which activate the coagulation cascade, resulting in a marked increase in thrombin generation and enhanced endothelial procoagulant activity (PCA) 67 .…”

“…Yu et al 67 demonstrated that UA at high levels can induce cytoskeletal structure disruption, transmembrane protein 16F (TMEM16F) activation, phosphatidylserine (PS) externalization, and MP shedding in HUVECs by increasing the levels of intracellular calcium and ROS. After MP shedding and PS externalization on ECs, binding sites are made available for the coagulation factors FXA and the prothrombinase complex, which activate the coagulation cascade, resulting in a marked increase in thrombin generation and enhanced endothelial procoagulant activity (PCA) 67 . Collectively, these studies indicate that the interaction between ECs and coagulation‐fibrinolytic systems is altered in HUA, increasing the risk of thromboembolic events.…”

Ferroptosis of Endothelial Cells Triggered by Erythrophagocytosis Contributes to Thrombogenesis in Uremia

“…1mL of blood was drawn into 3.8% sodium citrate tube from rats by cardiac puncture and mixed with 40μL kaolin. TEG analysis was performed according to Harmanpreet et al 25 , in brief, 340μL blood samples were placed into the prepared TEG cups with 20μL CaCl 2 (0.2M), and TEG analysis using a Haemoscope TEG 5000 Thrombelastograph Hemostasis Analyzer (Haemonetics Corp., Braintree, Massachusetts, USA) started immediately according to the manufacturer's instructions. The TEG run was stopped after all relevant parameters were assessed.…”