Abstract：
Recent investigation of a constitutively active ADAMTS13 variant (caADAMTS13) in murine models of acute ischaemic stroke (AIS) have revealed a potential anti-inflammatory mechanism of action contributing to its protective effect. However, it remains unclear whether these observations are a direct result of VWF proteolysis by caADAMTS13. We have implemented state of the art in vitro assays of neutrophil rolling and transmigration to quantify the impact of caADAMTS13 on these processes. Moreover, we have tested caADAMTS13 in two in vivo assays of neutrophil migration to confirm the impact of the treatment on the neutrophil response to sterile inflammation. Neutrophil rolling, over an interleukin-1β stimulated hCMEC/D3 monolayer, is directly inhibited by caADAMTS13, reducing the proportion of neutrophils rolling to 9.5 ± 3.8 % compared to 18.0 ± 4.5 % in untreated controls. Similarly, neutrophil transmigration recorded in real-time, was significantly suppressed in the presence of caADAMTS13 which reduced the number of migration events to a level like that in unstimulated controls (18.0 ± 4.5 and 15.8 ± 7.5 cells/mm2/h, respectively). Brain tissue from mice undergoing experimental focal cerebral ischaemia has indicated the inhibition of this process by caADAMTS13. This is supported by caADAMTS13\'s ability to reduce neutrophil migration into the peritoneal cavity in an ischaemia-independent model of sterile inflammation, with the VWF-dependent mechanism by which this occurs being confirmed using a second experimental stroke model. These findings will be an important consideration in the further development of caADAMTS13 as a potential therapy for AIS and other thromboinflammatory pathologies, including cardiovascular disease.
摘要：
最近在急性缺血性中风（AIS）的小鼠模型中对组成型活性ADAMTS13变体（caADAMTS13）的研究揭示了潜在的抗炎作用机制，有助于其保护作用。然而,目前尚不清楚这些观察结果是否是caADAMTS13VWF蛋白水解的直接结果.我们已经实施了嗜中性粒细胞滚动和迁移的现有技术体外测定,以量化caADAMTS13对这些过程的影响。此外，我们已经在中性粒细胞迁移的两项体内试验中测试了caADAMTS13,以确认治疗对中性粒细胞对无菌炎症反应的影响.中性粒细胞滚动，在白细胞介素-1β刺激的hCMEC/D3单层上，被caADAMTS13直接抑制,与未处理的对照中的18.0±4.5%相比,嗜中性粒细胞滚动的比例降低至9.5±3.8%。同样，实时记录中性粒细胞迁移，在存在caADAMTS13的情况下被显着抑制，这将迁移事件的数量减少到与未刺激对照相似的水平（18.0±4.5和15.8±7.5个细胞/mm2/h，分别)。来自经历实验性局灶性脑缺血的小鼠的脑组织表明caADAMTS13抑制了该过程。这是由caADAMTS13的能力，以减少中性粒细胞迁移到腹膜腔在一个独立的无菌炎症模型，使用第二个实验性中风模型证实了发生这种情况的VWF依赖性机制。这些发现将是进一步开发caADAMTS13作为AIS和其他血栓炎症病理的潜在疗法的重要考虑因素。包括心血管疾病.
