Department of Translational Neuroscience, Jing'an District Centre Hospital of Shanghai, State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Institutes of Brain Science, Fudan University, China (Y.H., L.C., Y.Z., X.G., Z.L., R.W., B.-Q.Z., W.F.).
Institute of Neuroscience and Third Affiliated Hospital, Zhengzhou University, China (Y.W.).
Stroke. 2024 Apr;55(4):1075-1085. doi: 10.1161/STROKEAHA.123.045991. Epub 2024 Mar 6.
Ischemic stroke is often accompanied by oxidative stress and inflammatory response, both of which work synergistically to exacerbate the disruption of the blood-brain barrier and ischemic brain injury. ALK (anaplastic lymphoma kinase), a cancer-associated receptor tyrosine kinase, was found to play a role in oxidative stress and inflammation. In this study, we investigated the role of ALK inhibition in a murine model of ischemic stroke.
Focal cerebral ischemia was induced by temporary occlusion of the right middle cerebral artery in mice with a filament. The ALK inhibitor alectinib was administered following the stroke. ALOX15 (arachidonic acid 15-lipoxygenase) was overexpressed by adenovirus injection. The immunohistochemistry, Western blot, oxidative stress, inflammation, blood-brain barrier leakage, infarct volume, and functional outcomes were determined.
We found that the expression of ALK was markedly increased in the neurovascular unit after cerebral ischemia. Treatment with the ALK inhibitor alectinib reduced the accumulation of reactive oxygen species, lipid peroxidation, and oxidative DNA, increased the vascular levels of antioxidant enzymes, inactivated the vascular NLRP3 (nucleotide-binding oligomerization domain-like receptor protein 3) inflammasome pathway, and reduced vascular inflammation (ICAM-1 [intercellular adhesion molecule-1] and MCP-1 [monocyte chemoattractant protein-1]) after ischemia. Moreover, alectinib reduced the loss of cerebrovascular integrity and blood-brain barrier damage, consequently decreasing brain infarction and neurological deficits. Furthermore, alectinib reduced stroke-evoked ALOX15 expression, whereas virus-mediated overexpression of ALOX15 abolished alectinib-dependent inhibition of oxidative stress and vascular inflammation, blood-brain barrier protection, and neuroprotection, suggesting the protective effects of alectinib for stroke may involve ALOX15.
Our findings demonstrated that alectinib protects from stroke by regulating ischemic signaling cascades and suggest that ALK may be a novel therapeutic target for ischemic stroke.
缺血性中风常伴有氧化应激和炎症反应,两者协同作用加剧血脑屏障破坏和缺血性脑损伤。ALK(间变性淋巴瘤激酶)是一种与癌症相关的受体酪氨酸激酶,被发现参与氧化应激和炎症反应。本研究旨在探讨ALK 抑制剂在缺血性中风小鼠模型中的作用。
通过线栓法阻塞小鼠右侧大脑中动脉造成局灶性脑缺血。中风后给予 ALK 抑制剂阿来替尼治疗。通过腺病毒注射过表达 ALOX15(花生四烯酸 15-脂氧合酶)。通过免疫组织化学、Western blot、氧化应激、炎症、血脑屏障渗漏、梗死体积和功能结果进行检测。
我们发现缺血后神经血管单元中 ALK 的表达明显增加。ALK 抑制剂阿来替尼治疗可减少活性氧、脂质过氧化和氧化 DNA 的积累,增加血管抗氧化酶水平,使血管 NLRP3(核苷酸结合寡聚化结构域样受体蛋白 3)炎症小体途径失活,减少血管炎症(ICAM-1[细胞间黏附分子-1]和 MCP-1[单核细胞趋化蛋白-1])。此外,阿来替尼减少了脑血管完整性的丧失和血脑屏障损伤,进而减少脑梗死和神经功能缺损。进一步研究发现,阿来替尼减少了中风引起的 ALOX15 表达,而病毒介导的 ALOX15 过表达则消除了阿来替尼依赖的抑制氧化应激和血管炎症、血脑屏障保护和神经保护作用,表明阿来替尼对中风的保护作用可能涉及 ALOX15。
我们的研究结果表明,阿来替尼通过调节缺血性信号级联反应来保护中风,提示 ALK 可能是缺血性中风的一个新的治疗靶点。