Center for Molecular Cardiology, University of Zurich, Schlieren, Switzerland.
First Clinic of Internal Medicine, Department of Internal Medicine, University of Genoa, Genoa, Italy.
Eur J Clin Invest. 2020 Feb;50(2):e13191. doi: 10.1111/eci.13191.
Endothelial cells regulate the formation of blood clots; thus, genes selectively expressed in these cells could primarily determine thrombus formation. Apold1 (apolipoprotein L domain containing 1) is a gene expressed by endothelial cells; whether Apold1 directly contributes to arterial thrombosis has not yet been investigated. Here, we assessed the effect of Apold1 deletion on arterial thrombus formation using an in vivo model of carotid thrombosis induced by photochemical injury.
Apold1 knockout (Apold1 ) mice and wild-type (WT) littermates underwent carotid thrombosis induced by photochemical injury, and time to occlusion was recorded. Tissue factor (TF) activity and activation of mitogen-activated protein kinases (MAPKs) and phosphatidyl-inositol-3 kinase (PI3K)/Akt pathways were analysed by colorimetric assay and Western blotting in both Apold1 and WT mice. Finally, platelet reactivity was assessed using light transmission aggregometry.
After photochemical injury, Apold1 mice exhibited shorter time to occlusion as compared to WT mice. Moreover, TF activity was increased in carotid arteries of Apold1 when compared to WT mice. Underlying mechanistic markers such as TF mRNA and MAPKs activation were unaffected in Apold1 mice. In contrast, phosphorylation of Akt was reduced in Apold1 as compared to WT mice. Additionally, Apold1 mice displayed increased platelet reactivity to stimulation with collagen compared with WT animals.
Deficiency of Apold1 results in a prothrombotic phenotype, accompanied by increased vascular TF activity, decreased PI3K/Akt activation and increased platelet reactivity. These findings suggest Apold1 as an interesting new therapeutic target in the context of arterial thrombosis.
内皮细胞调节血栓的形成；因此，这些细胞中选择性表达的基因可能主要决定血栓的形成。Apold1（载脂蛋白 L 结构域包含 1）是一种在内皮细胞中表达的基因；Apold1 是否直接导致动脉血栓形成尚未得到研究。在这里，我们使用光化学损伤诱导的颈动脉血栓形成的体内模型评估了 Apold1 缺失对动脉血栓形成的影响。
Apold1 敲除（Apold1 -/-）小鼠和野生型（WT）同窝仔鼠接受光化学损伤诱导的颈动脉血栓形成，并记录闭塞时间。通过比色法和 Western blot 分析 Apold1 和 WT 小鼠中组织因子（TF）活性以及丝裂原活化蛋白激酶（MAPKs）和磷脂酰肌醇-3 激酶（PI3K）/Akt 途径的激活。最后，使用透光比浊法评估血小板反应性。
光化学损伤后，Apold1 -/- 小鼠的闭塞时间较 WT 小鼠缩短。此外，与 WT 小鼠相比，Apold1 小鼠颈动脉 TF 活性增加。在 Apold1 -/- 小鼠中，TF mRNA 和 MAPKs 激活等潜在机制标志物不受影响。相比之下，与 WT 小鼠相比，Apold1 小鼠中的 Akt 磷酸化减少。此外，与 WT 动物相比，Apold1 小鼠对胶原刺激的血小板反应性增加。
Apold1 缺乏导致血栓形成表型，伴有血管 TF 活性增加、PI3K/Akt 激活减少和血小板反应性增加。这些发现表明 Apold1 是动脉血栓形成背景下一个有趣的新治疗靶点。
