CX3CL1 inhibits NLRP3 inflammasome-induced microglial pyroptosis and improves neuronal function in mice with experimentally-induced ischemic stroke
CX3CL1 inhibits NLRP3 inflammasome-induced microglial pyroptosis and improves neuronal function in mice with experimentally-induced ischemic stroke
复制标题
CX3CL1抑制NLRP3炎性小体诱导的小胶质细胞松弛和改善实验性缺血性卒中小鼠的神经功能
DOI：
10.1016/j.lfs.2022.120564
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发表时间：
2022-04-27
期刊：
影响因子：
6.1
通讯作者：
Lin, Yun
中科院分区：
文献类型：
--
作者：
Ge, Yangyang;Wang, Li;Lin, Yun
目的：中风是全球第二大死亡原因，是在脑血循环中断后发展起来的。趋化因子CX3CR1及其受体CX3CR1在缺血性脑卒中的病理生理中起重要作用。在本研究中，我们在体外和体内研究了CX3CL1对脑缺血的保护作用。主要方法：采用小鼠体内大脑中动脉闭塞(MCAO)/再灌注模型和体外BV2细胞氧糖剥夺/再氧合（OGD/R）模型。外源性重组CX3CL1 （rCX3CL1）在再灌注后1、3和5天(s)进入侧脑室或OGD/R后的细胞上清。采用免疫染色法、免疫印迹法和ELISA法对NLRP3炎性小体诱导的体内外焦亡进行评价。此外，观察MCAO后小鼠的神经功能缺损和梗死体积。关键发现：MCAO后CX3CL1表达下调。外源性rCX3CL1显著减少MCAO后小鼠的神经功能缺损和梗死灶。此外，外源性rCX3CL1抑制gsdmd依赖性小胶质细胞焦亡。这些作用进一步降低了NLRP3炎性体和nf - κ B信号的激活，并抑制了IL-1 β和IL-18的体外和体内表达。意义：这些结果表明，缺血损伤后外源性rCX3CL1给药对缺血性卒中后具有长期的神经保护作用。外源性rCX3CL1可抑制NLRP3炎症小体诱导的缺血性小胶质细胞焦亡。综上所述，我们的研究结果表明CX3CL1信号通路可以作为促进脑卒中后功能恢复的治疗靶点。
Aims: Stroke represents the second cause of mortality across the globe and develops following the interruption of cerebral blood circulation. The chemokine CX3CL1 and its receptor CX3CR1 play a fundamental role in the pathophysiology of ischemic stroke. In this study, we investigated the protective effect of CX3CL1 against ce-rebral ischemia both in vitro and in vivo. Main methods: We employed an in vivo mice model of middle cerebral artery occlusion(MCAO)/reperfusion and in vitro BV2 cells model of oxygen-glucose deprivation/re-oxygenation (OGD/R). Exogenous recombinant CX3CL1 (rCX3CL1) was administered into the lateral ventricle 1, 3 and 5 day(s) after reperfusion or in cell supernatant following OGD/R. Immunostaining, immunoblotting, and ELISA were performed to assess the NLRP3 inflammasome-induced pyroptosis both in vivo and in vitro. In addition, neurological deficits and infarct volume in mice were evaluated after MCAO. Key findings: The expression of CX3CL1 was downregulated after MCAO. Exogenous rCX3CL1 significantly reduced neurological deficits and infarct lesion in mice after MCAO. Moreover, exogenous rCX3CL1 inhibited GSDMD-dependent pyroptosis in microglia. Those effects further diminished NLRP3 inflammasome and NF-kappa B signaling activation, and also inhibited IL-1 beta and IL-18 expression both in vitro and in vivo. Significance: These results demonstrated that exogenous rCX3CL1 administration after the ischemic insult exerted a long-term neuroprotective effect on post-ischemic stroke. And exogenous rCX3CL1 could inhibit NLRP3 inflammasome-induced microglial pyroptosis under ischemic conditions. Collectively, our findings showed that CX3CL1 signaling pathway can serve as a therapeutic target for promoting the functional recovery after stroke.