NLRP3/Caspase-1/IL-1β炎症小体通路在小鼠主动脉夹层形成中的作用  

Role of NLRP3/Caspase-1/IL-1βinflammasome pathway in formation of aortic dissection in mice

作　　者：向军[1] 何玲[2] 许宏志[3] 梁维维[1] 彭泰峦[1] 魏蜀亮[1] XIANG Jun;HE Ling;XU Hongzhi;LIANG Weiwei;PENG Tailuan;WEI Shuliang(Department of Cardiac and Macrovascular Surgery,Affiliated Hospital of North Sichuan Medical College,Nanchong,Sichuan Province,637000,China;Department of Pediatrics/Neonatology,Affiliated Hospital of North Sichuan Medical College,Nanchong,Sichuan Province,637000,China;Surgery Center,Affiliated Hospital of North Sichuan Medical College,Nanchong,Sichuan Province,637000,China)

机构地区：[1]川北医学院附属医院心脏大血管外科,四川南充637000 [2]川北医学院附属医院儿科/新生儿科,四川南充637000 [3]川北医学院附属医院手术中心,四川南充637000

出　　处：《陆军军医大学学报》2024年第7期705-714,共10页Journal of Army Medical University

基　　金：四川省南充市科技局合作科研项目(22SXQT0007);川北医学院附属医院科研项目(2022LC018)。

摘　　要：目的探讨NLRP3/Caspase-1/IL-1β炎症小体通路在小鼠主动脉夹层形成中的作用及机制。方法50只3周龄雄性C57BL/6小鼠(体质量10~13 g)按随机抽样法分为对照组(n=10,无干预)、β-氨基丙腈(β-aminopropionitrile,BAPN)组[n=20,饮水中加入1 g/(kg·d)BAPN]、BAPN+MCC950组[n=20,饮水中加入1 g/(kg·d)BAPN、腹腔注射NLRP3抑制剂MCC95020 mg/(kg·d)]。记录各组小鼠饮水量、体质量、主动脉夹层发生率、夹层相关病死率。采用HE染色观察主动脉壁炎症浸润;弹力纤维(elastic Van Gieson,EVG)染色观察主动脉壁弹力纤维断裂。采用免疫荧光检测NLRP3、IL-1β、α-SMA、OPN的平均荧光强度。采用Western blot检测NLRP3、Caspase-1、ASC、IL-1β、α-SMA、OPN表达水平。结果对照组无主动脉夹层发生及死亡,BAPN组主动脉夹层发生率为80%,夹层相关病死率为35%,且主动脉血管壁弹力纤维断裂、明显炎症浸润。与对照组比较,BAPN组中NLRP3、Caspase-1、ASC、IL-1β蛋白表达增加,收缩型蛋白α-SMA减少而合成型蛋白OPN增加(P<0.05)。而给予NLRP3特异性抑制剂MCC950后,主动脉夹层发生率下降(80%vs 35%,P=0.004),夹层相关病死率下降(35%vs 15%,P=0.144),血管壁弹力纤维断裂、炎症浸润减少;与BAPN组比较,BAPN+MCC950组NLRP3、Caspase-1、ASC、IL-1β蛋白表达降低,收缩型蛋白α-SMA表达增加而合成型蛋白OPN表达降低(P<0.05)。结论小鼠主动脉夹层的发生与NLRP3/Caspase-1/IL-1β炎症小体通路活化有关,而NLRP3抑制剂MCC950能减少主动脉夹层发生,具有血管保护作用。Objective To investigate the role and mechanism of NLRP3/Caspase-1/IL-1βinflammasome pathway in the formation of aortic dissection in mice.Methods Fifty male C57BL/6 mice(3 weeks old,body weight 10~13 g)were divided into control group(n=10,normal diet),β-aminopropionitrile(BAPN)group[n=20,drink water containing 1 g/(kg·d)BAPN],and BAPN+MCC950 group[n=20,drink water containing 1 g/(kg·d)BAPN and intraperitoneal injection of 20 mg/(kg·d)NLRP3 inhibitor,MCC950]by random sampling.Water intake,body weight,incidence of aortic dissection and aortic dissection-related mortality were recorded.The inflammatory infiltration in the aorta was observed with HE staining,elastic fiber breakage was observed by elastic Van Gieson(EVG)staining,average fluorescence intensity of NLRP3,IL-1β,α-SMA and OPN was detected by immunofluorescence assay,and protein expression levels of NLRP3,Caspase-1,ASC,IL-1β,α-SMA and OPN were measured with Western blotting.Results No aortic dissection or death was observed in the control group.The BAPN group had an incidence of aortic dissection of 80%,aortic dissection-related mortality of 35%,and obvious broken elastic fibers and inflammatory infiltrate in the aortic wall,and increased expression levels of NLRP3,Caspase-1,ASC and IL-1β,decreased contractileα-SMA and increased synthetic protein OPN when compared with the control group(P<0.05).While MCC950 treatment decreased the incidence of aortic dissection(80%vs 35%,P=0.004)and aortic dissection-related mortality(35%vs 15%,P=0.144),alleviated the broken elastic fibers and inflammatory infiltrate in the aortic wall,and down-regulated the expression of NLRP3,Caspase-1,ASC and IL-1β,enhanced contractileα-SMA and decreased the synthetic protein when compared with the BAPN group(P<0.05).Conclusion The occurrence of aortic dissection in mice is associated with activation of NLRP3/Caspase-1/IL-1βinflammasome pathway.NLRP3 inhibitor,MCC950,can reduce the occurrence of aortic dissection and show a protective effect on blood vessels.

关 键 词：主动脉夹层 炎症 NLRP3 信号通路 

分 类 号：R363.21[医药卫生—病理学] R392.13[医药卫生—基础医学] R543.16