MCC950抑制NLRP3炎性小体活性改善阿尔茨海默病模型小鼠血脑屏障功能研究  

Inhibition of NLRP3 Inflammasome Activity by MCC950 Improves Blood-Brain Barrier Function in Alzheimer's Disease Model Mice

作　　者：董笑博 安阳[1] 杨彩瑜 房小楠 贾冬[1] 齐越[1] DONG Xiaobo;AN Yang;YANG Caiyu;FANG Xiaonan;JIA Dong;QI Yue(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847

出　　处：《辽宁中医药大学学报》2023年第9期57-62,共6页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：辽宁省科技厅项目(2019-ZD-0843);沈阳市科技计划项目(19-112-4-072)。

摘　　要：目的研究核苷酸结合域样受体蛋白3(NLRP3)炎性小体抑制剂MCC950对侧脑室注射淀粉样蛋白(Aβ25-35)所致的阿尔茨海默病(AD)模型小鼠血脑屏障(BBB)结构和功能的影响。方法将小鼠随机分为假手术组、模型组、盐酸多奈哌齐组(1.3 mg/kg)和MCC950组(10 mg/kg),除假手术组外,其余各组侧脑室注射Aβ25-35制作小鼠AD模型,造模后第2天,各给药组按相应剂量连续给药21 d,每日1次,假手术组给予等量0.9%氯化钠溶液;于第15天进行Y迷宫实验,第16~21天进行Morris水迷宫实验;行为学实验后进行伊文思蓝渗漏实验并取材;ELISA试剂盒及免疫荧光染色法检测小鼠脑组织中NLRP3水平,Western blot法检测小鼠脑组织海马及皮质中紧密连接蛋白-5(claudin-5)、闭合蛋白(occludin)、闭锁小带蛋白-1(ZO-1)、凋亡相关斑点样蛋白(ASC)、半胱天冬氨酸酶(caspase-1)、白细胞介素-18(IL-18)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、p53上调凋亡调节因子(PUMA)、抑癌因子p53、核转录因子κB(NF-κB)的表达情况。结果与假手术组相比,模型组小鼠自发交替反应率下降,水迷宫的游泳潜伏期较长,穿越平台次数及目标象限路程减少,脑内伊文思蓝渗透率显著增加,NLRP3表达增多,claudin-5、occludin、ZO-1表达明显减少;ASC、caspase-1、IL-1β、IL-18、TNF-α、PUMA、p53、NF-κB表达明显增加。与模型组比较,给药组小鼠自发交替反应率升高,水迷宫潜伏期较短,穿越平台次数及目标象限路程增加,小鼠脑内伊文思蓝含量降低,NLRP3表达明显减少,claudin-5、occludin、ZO-1明显增加;ASC、caspase-1、IL-1β、IL-18、TNF-α、PUMA、p53、NF-κB表达明显减少。结论MCC950可通过PUMA/NF-κB和PUMA/p53信号通路抑制NLRP3炎性小体活性来改善AD模型小鼠血脑屏障功能。Objective To investigate the effect of NLRP3 inflammasome inhibitor MCC950 on the structure and function of BBB in AD model mice induced by lateral ventricular injection of Aβ25-35-Methods Mice were randomly divided into sham-operated group,model group,donepezil hydrochloride group(1.3 mg/kg)and MCC950 group(10 mg/kg);except for the sham-operated group,each group was injected with Aβ25-35 in the lateral ventricle to make mice AD models;on the second day after modeling,each administration group was given the corresponding dose for 21 consecutive days,once daily,and the sham-operated group was given equal amount of physiological saline;The Y-maze experiment was performed on day 15,and the Morris water-maze experiment was performed on days 16-21;After the behavioral experiments,Evans blue leakage test was performed and the material was taken;the level of NLRP3 in mouse brain tissue wes detected by ELISA kit and immunofluorescence staining,Western blot measured the expression of claudin-5,occludin,ZO-1,ASC,caspase-1,IL-18,IL-1β,TNF-α,PUMA,p53 and NF-κB in hippocampus and cortex of mice brain tissue.Results Compared with the sham-operated group,mice in the model group showed decreased spontaneous alternating response rate,longer swimming latency in the water maze,decreased the times of crossing the platform and the distance traveled in the target quadrant,significantly increased intracerebral Evans blue infiltration,and increased NLRP3 expression.The expressions of claudin-5,occludin and ZO-1 was significantly decreased,and the expressions of ASC,caspase-1,IL-1β,IL-18,TNF-α,PUMA,p53 and NF-κB were significantly increased.Compared with the model group,the mice in the two administration groups had higher spontaneous alternating response rate,shorter swimming latency in the water maze,increased times of platform crossings and distance to the target quadrant,decreased Evans blue content in the mouse brain,significantly decreased NLRP3 expression,significantly increased the expressions of claudin-5,occludin,ZO-1;an

关 键 词：MCC950 NLRP3炎性小体 阿尔茨海默病 血脑屏障 

分 类 号：R741[医药卫生—神经病学与精神病学]