Yan Meishan, Xu Minghui, Li Zhanni, An Yao, Wang Zelong, Li Shuli, Chen Yingli, Xia Yanshi, Wang Liqiu, Wang Longlong, Ji Shuting, Dong Weijun, Shi Jialan, Gao Chunyan
Department of Medical Laboratory Science and Technology, Harbin Medical University-Daqing, Daqing, China.
Department of Anesthesiology, Daqing Oil Field General Hospital, Daqing, China.
Blood Cells Mol Dis. 2022 May 7;96:102666. doi: 10.1016/j.bcmd.2022.102666.
The link between hyperuricemia (HUA) and the risk of venous thromboembolism (VTE) has been well established. However, the mechanisms of thrombus generation and the effect of HUA on procoagulant activity (PCA) of erythrocytes remain unclear no matter in uremia or hyperuricemia. Here, phosphatidylserine (PS) exposure, microparticles (MPs) release, cytosolic Ca, TMEM16F expression, reactive oxygen species (ROS) and lipid peroxidation of erythrocyte were detected by flow cytometer. PCA was assessed by coagulation time, purified coagulation complex and fibrin production assays. The fibrin formation was observed by scanning electron microscopy (SEM). We found that PS exposure, MPs generation, TMEM16F expression and consequent PCA of erythrocyte in HUA patients significantly increased compared to those in healthy volunteers. Furthermore, high UA induced PS exposure, and MPs release of erythrocyte in concentration and time-dependent manners in vitro, which enhanced the PCA of erythrocyte and was inhibited by lactadherin, a PS inhibitor. Additionally, using SEM, we also observed compact fibrin clots with highly-branched networks and thin fibers supported by red blood cells (RBCs) and RBC-derived MPs (RMPs). Importantly, we demonstrated UA enhanced the production of ROS and lipid peroxidation and reduced the generation of glutathione (GSH) of erythrocyte, which enhanced TMEM16F activity and followed PS externalization and RMPs formation. Collectively, these results suggest that Ca-dependent TMEM16F activation may be responsible for UA-induced PS exposure and MPs release of RBC, which thereby contribute to the prothrombotic risk in HUA.
高尿酸血症（HUA）与静脉血栓栓塞症（VTE）风险之间的联系已得到充分证实。然而，无论是在尿毒症还是高尿酸血症中，血栓形成的机制以及HUA对红细胞促凝活性（PCA）的影响仍不清楚。在此，通过流式细胞仪检测红细胞的磷脂酰丝氨酸（PS）暴露、微粒（MPs）释放、胞质钙、TMEM16F表达、活性氧（ROS）和脂质过氧化。通过凝血时间、纯化凝血复合物和纤维蛋白生成试验评估PCA。通过扫描电子显微镜（SEM）观察纤维蛋白形成。我们发现，与健康志愿者相比，HUA患者红细胞的PS暴露、MPs生成、TMEM16F表达及随后的PCA显著增加。此外，高尿酸在体外以浓度和时间依赖性方式诱导红细胞PS暴露和MPs释放，增强了红细胞的PCA，并被PS抑制剂乳黏素抑制。此外，使用SEM，我们还观察到由红细胞（RBCs）和红细胞衍生的MPs（RMPs）支持的具有高度分支网络和细纤维的紧密纤维蛋白凝块。重要的是，我们证明尿酸增强了红细胞ROS的产生和脂质过氧化，并减少了谷胱甘肽（GSH）的生成，这增强了TMEM16F活性，随后导致PS外化和RMPs形成。总的来说，这些结果表明，钙依赖性TMEM16F激活可能是尿酸诱导红细胞PS暴露和MPs释放的原因，从而导致HUA中的血栓形成风险增加。
