NLRP3炎症小体的活化水平与急性缺血性脑卒中患者认知功能改变的关系

Relationship between NLRP3 activation level of inflammasome and the change of cognitive functions in patients with acute ischemic stroke

目的探讨Nod样受体热蛋白结构域相关蛋白3(Nod-like receptor pyrin domain containing 3,NLRP3)炎症小体的活化水平与急性缺血性脑卒中患者认知功能改变的关系。方法选择2018年10月至2020年7月神经内科收治的88例急性缺血性脑卒中患者(病例组)及100例体检者(对照组)作为研究对象。采集病例组及对照组外周静脉血,离心分离外周血单核淋巴细胞(peripheral blood mononuclear cells,PBMCs),采用Western blot检测NLRP3、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD,ASC)、Caspase-1、白细胞介素-1β(interleukin-1β,IL-1β)蛋白表达,采用蒙特利尔认知功能评估量表(Montreal cognitive assessment,MoCA)对脑卒中患者认知功能进行检查,比较病例组及对照组NLRP3、ASC、Caspase-1、IL-1β表达水平差异。所有数据采用SPSS 19.0进行统计分析,Pearson相关分析检验NLRP3、ASC、Caspase-1、IL-1β表达与MoCA评分的相关性,Logistic多因素回归分析NLRP3、ASC、Caspase-1、IL-1β表达与认知功能障碍的关系。结果(1)Western blot结果显示,病例组患者PBMCs细胞中NLRP3、ASC、Caspase-1、IL-1β蛋白表达高于对照组(均P<0.05)。(2)合并高血压、高脂血症、美国国立卫生院卒中量表(NIHSS)评分≥8分脑卒中患者NLRP3表达量显著高于无高血压、无高脂血症、NIHSS评分<8分的患者(P<0.05)。(3)病例组中认知功能障碍发生率为34.09%(30/88);认知障碍组及非认知障碍组MoCA得分分别为20(24,28)分、27(26,28)分,组间比较差异具统计学意义(P<0.05)。(4)Pearson相关分析显示,PBMCs细胞NLRP3、ASC、Caspase-1、IL-1β表达与MoCA评分呈负相关(r=-0.426,-0.396,-0.417,-0.320,均P<0.05)。(5)Logistic回归分析显示,合并高脂血症、NIHSS评分、额颞叶梗死、NLRP3表达是认知功能障碍发生的影响因素(均P<0.05)。结论急性缺血性脑卒中患者存在NLRP3炎症小体的活化,且其活化程度与病情及卒中后认知功能损伤的发生密切相关,靶向抑制或调控NLRP3炎症小体的活化可成为急性缺血性脑卒中神经保护的新思路。

Objective To investigate the relationship between the activation level of Nod-like receptor pyrin domain-containing 3(NLRP3)inflammasome and the change of cognitive functions in patients with acute ischemic stroke.Methods A total of 88 patients with acute ischemic stroke in Department of Neurology from October 2018 to July 2020 were selected as case group and 100 healthy physical examinees were selected as control group.Peripheral blood of the case group and the control group was collected,and peripheral blood mononuclear cells(PBMCs)were isolated by centrifugation.Then the NLRP3,apoptosis-associated speck-like protein containing a CARD(ASC),Caspase-1 and interleukin-1β(IL-1β)expression were detected by Western blot.The cognitive function of patients with acute ischemic stroke was detected by Montreal Cognitive Assessment(MoCA).The differences in expression levels of NLRP3,ASC,Caspase-1 and IL-1βwere compared between the case group and the control group.Pearson correlation analysis was used to analyze the correlation between expression levels of NLRP3,ASC,Caspase-1,IL-1βand MoCA score.Logistic multivariate regression was used to analyze the relationship between expression levels of NLRP3,ASC,caspase-1,IL-1βand the cognitive dysfunction.Results(1)Western blot results showed that NLRP3,ASC,Caspase-1 and IL-1βexpressions in PBMCs cells in the case group were higher than those in the control group(all P<0.05).(2)The expression level of NLRP3 in stroke patients with hypertension,hyperlipidemia,National Institutes of Health Stroke Scale(NIHSS)score≥8 points was significantly higher than that in patients without hypertension,hyperlipidemia and NIHSS score<8 points(P<0.05);(3)The incidence of cognitive dysfunction in the case group was 34.09%(30/88).The MoCA scores of the cognitive dysfunction group and the non-cognitive dysfunction group were 20(24,28)and 27(26,28)points respectively,and the difference between the groups was statistically significant(P<0.05);(4)Pearson correlation analysis showed that NLRP3,ASC,caspase-1 and IL-1βexpression in PBMCs cells were negatively correlated with MoCA scores(r=-0.426,-0.396,-0.417,-0.320 respectively,all P<0.05).(5)Logistic regression analysis showed that hyperlipidemia,NIHSS scores,frontotemporal lobe infarction,and NLRP3 expression were the influencing factors for the occurrence of cognitive dysfunction(all P<0.05).Conclusion Patients with acute ischemic stroke have high activated NLRP3 inflammasome,and its activation degree is closely related to the condition and the occurrence of cognitive dysfunction after stroke.Targeted inhibition or regulation of NLRP3 inflammasome activation may become a new idea of neuroprotection for acute ischemic stroke.