Caspase-1 inhibition prevents neuronal death by targeting the canonical inflammasome pathway of pyroptosis in a murine model of cerebral ischemia
Caspase-1 inhibition prevents neuronal death by targeting the canonical inflammasome pathway of pyroptosis in a murine model of cerebral ischemia
复制标题
Caspase-1 抑制通过靶向小鼠脑缺血模型中细胞焦亡的经典炎性体途径来防止神经元死亡。
DOI：
10.1111/cns.13384
复制
发表时间：
2020-04-28
影响因子：
5.5
通讯作者：
Wang, Wei
中科院分区：
文献类型：
--
作者：
Li, Jia;Hao, Jia-Huan;Wang, Wei
目的焦亡在缺血性脑卒中中的作用尚不明确。因此，我们使用特异性焦亡抑制剂Vx765作为脑卒中小鼠模型的实验干预靶点。方法564只C57BL/6小鼠在缺血1小时后进行光凝治疗并灌胃Vx765。随后，我们使用免疫荧光（IF）和Western blot （WB）分析评估了Gasdermin D （GSDMD）、炎性小体、caspase-1和白细胞介素-1 β (IL-1 β)的表达。我们还用透射电子显微镜（TEM）检查了皮层神经元的超微结构变化，并用磁共振成像（MRI）动态测量了梗死体积。此外，我们通过改良的神经严重程度评分、rotarod测试和Treadscan来评估神经功能缺损。结果缺血后第1天和第3天，大鼠缺血周围区GSDMD和成孔亚基GSDMD p30的表达明显升高。脑卒中后第3天，神经元血浆、细胞核和线粒体膜出现超微结构损伤。损伤后第1天和第3天炎症小体、caspase-1和IL-1 β的表达也升高。vx765治疗组和对照组的平均梗死面积差异有统计学意义（14.36 vs 21.52 mm）；12.34 vs 18.56 mm(3)；4.13 vs 10.06 mm(3)；术后第1天、第3天、第7天P < 0.05)。通过一系列行为测试评估，Vx765处理的小鼠表现出更好的运动恢复，并且具有更好的神经元存活，这可归因于焦亡抑制，如效应蛋白GSDMD、炎症小体、caspase-1和IL-1 β的下调表达所示。此外，Vx765对缺血损伤后的神经元膜结构有保护作用。结论焦亡是急性缺血性脑卒中神经元死亡的重要途径。Vx765是一种低分子量药物，在癫痫临床试验中被证明是安全的，通过靶向焦亡的典型炎性体途径，对脑缺血具有潜在的治疗价值。
Aims The involvement of pyroptosis in ischemic stroke remains to be established. Therefore, we used the specific pyroptosis inhibitor Vx765 as an experimental intervention target in a murine model of stroke.Methods A total of 564 C57BL/6 mice were subjected to photothrombotic procedures and treated via gavage with Vx765 at 1-hour post-ischemia. We subsequently assessed the expression of Gasdermin D (GSDMD), inflammasomes, caspase-1, and interleukin-1 beta (IL-1 beta) using immunofluorescence (IF) and Western blot (WB) analyses. We also examined ultrastructural changes of cortical neurons with transmission electron microscopy (TEM) and measured infarct volumes dynamically by magnetic resonance imaging (MRI). Moreover, we evaluated the neurologic deficits by modified neurological severity scores, the rotarod test, and Treadscan.Results Elevated expression of GSDMD and GSDMD p30, the pore-forming subunit, was evident in the peri-ischemic region on days one and three post-ischemia. The neuronal plasma, nuclear, and mitochondrial membranes showed ultrastructural damage at day three post-stroke. Elevated expression of inflammasomes, caspase-1, and IL-1 beta was also present on days one and three post-injury. There were significant differences between Vx765-treated and vehicle groups in mean infarct volumes (14.36 vs 21.52 mm(3); 12.34 vs 18.56 mm(3); 4.13 vs 10.06 mm(3); P < .05 at day one, three, and seven post-surgery, respectively). Mice treated with Vx765 showed better motor recovery as assessed by serial behavior tests and had better neuronal survival, which was attributable to pyroptosis inhibition, as illustrated by downregulated expression of the effector protein GSDMD, inflammasomes, caspase-1, and IL-1 beta. Besides, treatment with Vx765 preserved neuronal membrane structures after the ischemic injury.Conclusions Pyroptosis emerges as an important pathway for neuronal death in an acute ischemic stroke. Vx765, a low molecular weight drug that has proven safe in clinical epilepsy trials, has potential therapeutic value for cerebral ischemia by targeting the canonical inflammasome pathway of pyroptosis.