基于NLRP3炎性小体探讨清心开窍方对APP/PS1双转基因小鼠神经炎症的影响  

The effect of Qingxin Kaiqiao Formula on neuroinflammation in APP/PS1 double transgenic mice based on NLRP3 inflammasome

作　　者：王柳莺 赖晓晓 刘硕 徐露婷 沈燕 胡海燕[1,2] WANG Liuying;LAI Xiaoxiao;LIU Shuo;XU Luting;SHEN Yan;HU Haiyan(Department of Traditional Chinese Medicine,the Second Affiliated Hospital of Wenzhou Medical University,Wenzhou 325027,China;The Second Clinical Medical College,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]温州医科大学附属第二医院中医科,浙江温州325027 [2]温州医科大学第二临床医学院,浙江温州325035

出　　处：《温州医科大学学报》2023年第6期431-439,共9页Journal of Wenzhou Medical University

基　　金：国家自然科学基金项目(81573896);浙江省自然科学基金项目(LY15H270016)。

摘　　要：目的:研究清心开窍方对阿尔兹海默病(AD)模型小鼠神经性炎症和认知能力的影响,并探讨与核苷酸结合寡聚域样受体蛋白3(NLRP3)炎性小体相关通路的关联。方法:将APP/PS1双转基因小鼠随机分为模型组、多奈哌齐治疗组[1.67 mg/(kg·d)]、清心开窍方治疗组[低剂量组:4.75 mg/(kg·d);中剂量组:9.5 mg/(kg·d);高剂量组:19 mg/(kg·d)],每组10只。SPF级的雄性C57BL/6J小鼠作为对照组。Morris水迷宫实验测试定向导航和空间感知能力;HE和Nissl染色测定海马区的细胞和尼氏体数目的变化;蛋白质印迹(Western blot)和定量实时荧光聚合酶链反应(RT-q PCR)测定NLRP3、Caspase-1、Aβ、IL-1β和IL-6的表达。结果:与对照组相比,模型组小鼠的平均逃逸潜伏期明显增加(P<0.01),通过平台次数和在平台所在目标象限停留时间明显减少(P<0.01),海马区神经元数量减少,不规则,尼氏体数量减少,染色颜色浅,NLRP3、Caspase-1、Aβ、IL-1β、IL-6表达明显增加(P<0.01);与模型组相比,多奈哌齐治疗组和清心开窍方治疗组逃避潜伏期均明显缩短,穿越平台次数及在平台所在象限停留时间明显增多(P<0.05或P<0.01),治疗组小鼠神经细胞排列更为紧密有序,形态更为完整,尼氏体数目明显增加,颜色加深,NLRP3、Caspase-1、Aβ、IL-1β和IL-6的表达明显减少(P<0.05或P<0.01)。结论:清心开窍方可能通过抑制NLRP3炎症通路减轻神经炎症改善APP/PS1小鼠的Aβ病理变化以及学习和记忆障碍。Objective:To investigate the effect of the Qingxin Kaiqiao Formula on neuroinflammation and cognitive performance in mice with Alzheimer’s disease(AD)model and to explore its association with NLRP3 inflammasome-related pathways.Methods:APP/PS1 double transgenic mice were randomly divided as model group,Aricept treatment group(1.67 mg·kg^(-1)·d^(-1)),and Qingxin Kaiqiao Formula treatment group(low dose group:4.75 mg·kg^(-1)·d^(-1);middle dose group:9.5 mg·kg^(-1)·d^(-1);high dose group:19 mg·kg^(-1)·d^(-1)),with ten mice in each group.SPF-grade male C57BL/6J mice served as the control group.The Morris water maze experiment was performed to test the ability of directional navigation and spatial perception;HE and Nissl staining were performed to determine changes in the number of cells and Nissl bodies in the hippocampal region;Western blot and real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)were used to determine the expression of NLRP3,Caspase-1,Aβ,IL-1β,and IL-6.Results:Compared with the control group,mice in the model group showed significantly increased mean escape latency(P<0.01),significantly decreased number of passage through the plateau shorter residence time in the target quadrant where the plateau was located(P<0.01),decreased number of neurons in the hippocampal region,irregularity,decreased number of Nissl bodies,lighter staining color,and obviously increased expression of NLRP3,caspase-1,Aβ,IL-1βand IL-6(P<0.01).Compared with the model group,both the Aricept treatment group and the Qingxin Kaiqiao Fang-treated group showed visibly shorter escape latency,significantly greater number of crossing platforms longer residence time in the quadrant where the plateau was located(P<0.05 or P<0.01);and the neuronal cells in the treated group were more closely arranged and more intact morphologically,apparently higher number of Nissl bodies deeper color,and significantly reduced NLRP3,Caspase-1,Aβ,IL-1βand IL-6 expression(P<0.05 or P<0.01).Conclusion:Qingxin Kaiqiao Formula m

关 键 词：阿尔兹海默病 清心开窍方 APP/PS1模型小鼠 NLRP3炎性小体 

分 类 号：R259[医药卫生—中西医结合]