Lu Danli, Hu Mengyan, Zhang Bingjun, Lin Yinyao, Zhu Qiang, Men Xuejiao, Lu Zhengqi, Cai Wei
Department of Neurology, Mental and Neurological Disease Research Center, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
Center of Clinical Immunology, Mental and Neurological Disease Research Center, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
Front Neurol. 2021 Apr 22;12:621555. doi: 10.3389/fneur.2021.621555. eCollection 2021.
The inflammasome represents a highly pro-inflammatory mechanism. It has been identified that inflammasome was activated after ischemic stroke. However, the impact of inflammasomes on stroke outcomes remains contradictory. The participating molecules and the functioning arena of post-stroke inflammasome activation are still elusive. In the present study, blood samples from stroke patients were collected and analyzed with flow cytometry to evaluate the correlation of inflammasome activation and stroke outcomes. A stroke model was established using male C57/Bl6 mice with transient middle cerebral artery occlusion (tMCAO, 1 h). The dynamics of inflammasome components, cell type, and location of inflammasome activation and the therapeutic effects of inhibiting post-stroke inflammasome executors were evaluated. We found that a high level of inflammasome activation might indicate detrimental stroke outcomes in patients and mice models. Post-stroke inflammasome activation, especially NLRP3, cleaved Caspase-1, cleaved Caspase-11, IL-1β, IL-18, and GSDMD, peaked at 3-5 days and declined at 7 days with the participation of multiple components in mice. Macrophage that infiltrated into the ischemic lesion was the main arena for post-stroke inflammasome activation among myeloid cells according to the data of mice. Among all the members of the Caspase family, Caspase-1 and -11 served as the main executing enzymes. Inhibiting Caspase-1/-11 signaling efficiently suppressed DAMPs-induced macrophage inflammasome activation and displayed neuroprotection to stroke models including infarct size (Control: 48.05 ± 14.98; Cas1.i: 19.34 ± 12.21; Cas11.i: 21.43 ± 14.67, < 0.001) and neurological deficit score (0 d-Control: 2.20 ± 0.63; 0 d-Cas1.i: 2.20 ± 0.63; 0 d-Cas11.i: 2.20 ± 0.63; 1 d-Control: 2.50 ± 0.53; 1 d-Cas1.i: 1.50 ± 0.71; 1 d-Cas11.i: 2.00 ± 0.67; 2 d-Control: 2.30 ± 0.48; 2 d-Cas1.i: 1.30 ± 0.48; 2 d-Cas11.i: 1.50 ± 0.53; 3 d-Control: 2.00 ± 0.67; 3 d-Cas1.i: 1.20 ± 0.42; 3 d-Cas11.i: 1.30 ± 0.48, < 0.001). Taken together, inflammasome activation played a detrimental role in stroke pathology. Targeting post-stroke inflammasome executing enzymes fitting in the dynamics of macrophages might obtain potential and efficient therapeutic effects.
炎性小体代表一种高度促炎机制。已确定缺血性中风后炎性小体被激活。然而，炎性小体对中风结局的影响仍存在矛盾。中风后炎性小体激活的参与分子和作用场所仍不清楚。在本研究中，收集了中风患者的血样并用流式细胞术进行分析，以评估炎性小体激活与中风结局的相关性。使用雄性C57/Bl6小鼠建立短暂性大脑中动脉闭塞（tMCAO，1小时）的中风模型。评估了炎性小体成分的动态变化、细胞类型、炎性小体激活的位置以及抑制中风后炎性小体执行者的治疗效果。我们发现，炎性小体的高水平激活可能表明患者和小鼠模型中中风结局不佳。中风后炎性小体激活，尤其是NLRP3、裂解的半胱天冬酶-1、裂解的半胱天冬酶-11、白细胞介素-1β、白细胞介素-18和Gasdermin D，在第3至5天达到峰值，并在第7天下降，小鼠中有多种成分参与。根据小鼠数据，浸润到缺血性病变中的巨噬细胞是骨髓细胞中中风后炎性小体激活的主要场所。在所有半胱天冬酶家族成员中，半胱天冬酶-1和-11是主要的执行酶。抑制半胱天冬酶-1/-11信号可有效抑制损伤相关分子模式诱导的巨噬细胞炎性小体激活，并对中风模型显示出神经保护作用，包括梗死面积（对照组：48.05±14.98；Cas1.i组：19.34±12.21；Cas11.i组：21.43±14.67，<0.001）和神经功能缺损评分（0天-对照组：2.20±0.63；0天-Cas1.i组：2.20±0.63；0天-Cas11.i组：2.20±0.63；1天-对照组：2.50±0.53；1天-Cas1.i组：1.50±0.71；1天-Cas11.i组：2.00±0.67；2天-对照组：2.30±0.48；2天-Cas1.i组：1.30±0.48；2天-Cas11.i组：1.50±0.53；3天-对照组：2.00±0.67；3天-Cas1.i组：1.20±0.42；3天-Cas11.i组：1.30±0.48，<0.001）。综上所述，炎性小体激活在中风病理过程中起有害作用。针对符合巨噬细胞动态变化的中风后炎性小体执行酶可能会获得潜在而有效的治疗效果。
