Inhibition of the inflammasome complex reduces the inflammatory response after thromboembolic stroke in mice
Inhibition of the inflammasome complex reduces the inflammatory response after thromboembolic stroke in mice
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DOI：
10.1038/jcbfm.2008.143
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发表时间：
2009-03-01
影响因子：
6.3
通讯作者：
Dietrich, W. Dalton
中科院分区：
文献类型：
--
作者：
Abulafia, Denise P.;Vaccari, Juan Pablo de Rivero;Dietrich, W. Dalton
炎症是脑缺血和中风的发病机制的主要贡献者。在外周免疫应答中，半胱天冬酶-1活化涉及称为炎性体的大分子复合物的形成。我们确定是否核苷酸结合，亮氨酸丰富的重复序列，pyrin结构域包含1（NLRP 1），分子平台组成的半胱氨酸蛋白酶-1，凋亡相关的斑点样蛋白含有半胱氨酸蛋白酶激活募集结构域（ASC），和NLRP 1，在正常和缺血后的大脑中表达。小鼠经历血栓栓塞性中风以研究炎性小体的形成和随后下游炎性反应的激活。Western blot分析显示，在中风后24小时，白细胞介素（IL）、IL-1 β和IL-18的表达和活化。分子排阻层析和免疫共沉淀分析显示NLRP 1、ASC、caspase-1和X连锁凋亡抑制蛋白（XIAP）之间存在蛋白关联。缺血后，免疫组织化学分析显示神经元，星形胶质细胞和小胶质细胞/巨噬细胞中的炎性蛋白。通过干扰炎性小体活化，从而在用针对NLRP 1的中和抗体治疗的小鼠中缺血后导致细胞因子水平降低，显示了炎性小体作为炎症靶点的潜力。这些研究结果表明，炎性小体复合物形成局灶性脑缺血后，可能是一种新的治疗靶点，以减少缺血后炎症的有害后果。
Inflammation is a major contributor to the pathogenesis of cerebral ischemia and stroke. In the peripheral immune response, caspase-1 activation involves the formation of a macromolecular complex termed the inflammasome. We determined whether nucleotide-binding, leucine-rich repeat, pyrin domain containing 1 (NLRP1), molecular platform consisting of capase-1, apoptosis-associated speck-like protein containing a caspase-activating recruitment domain (ASC), and NLRP1, is expressed in the normal and postischemic brain. Mice underwent thromboembolic stroke to investigate the formation of the inflammasome and subsequent activation of downstream inflammatory responses. Western blot analysis showed expression and activation of interleukin (IL) IL-1 beta and IL-18 at 24 h after stroke. Size-exclusion chromatography and coimmunoprecipitation analysis showed protein association between NLRP1, ASC, caspase-1, and the X-linked inhibitor of apoptosis protein (XIAP). After ischemia, immunohistochemical analysis revealed inflammasome proteins in neurons, astrocytes, and microglia/macrophages. The potential of the inflammasome as an antiinflammatory target was showed by interference of inflammasome activation resulting in reduced cytokine levels in mice treated after ischemia with a neutralizing antibody against NLRP1. These findings show that the inflammasome complex forms after focal brain ischemia and may be a novel therapeutic target for reducing the detrimental consequences of postischemic inflammation.