黑逍遥散对APP/PS1小鼠神经炎症及NLRP3/Caspase-1/GSDMD信号通路的影响  

Effect of Hei Xiaoyaosan on Neuroinflammation and NLRP3/Caspase-1/GSDMD Signaling Pathway in APP/PS1 Mice

作　　者：周君[1,2] 李明成 吕育洁 孟志鹏 胡韵韵 王虎平 ZHOU Jun;LI Mingcheng;LYU Yujie;MENG Zhipeng;HU Yunyun;WANG Huping(Gansu Engineering Laboratory for New Products of Traditional Chinese Medicine(TCM),Gansu Key Laboratory of TCM Excavation and Innovative Transformation,Gansu University of Chinese Medicine,Lanzhou 730000,China;Xichang Hospital of TCM,Xichang 615000,China;Qujing 69 Hospital,China RongTong Medical Healthcare Group Co.,Ltd.,Qujing 655000,China;Xingtai Hospital of TCM,Xingtai 054000,China;Zhengzhou TCM Hospital Affiliated to Henan University of Chinese Medicine,Zhengzhou 450007,China;Guizhou Health Vocational College,Tongren 554300,China)

机构地区：[1]甘肃中医药大学甘肃省中药新产品创制工程实验室,甘肃省中医方药挖掘与创新转化重点实验室,兰州730000 [2]西昌市中医医院,四川西昌615000 [3]中国融通医疗健康集团有限公司,曲靖六十九医院655000 [4]邢台市中医医院,邢台054000 [5]河南中医药大学附属郑州市中医院,郑州450007 [6]贵州健康职业学院,贵州铜仁554300

出　　处：《中国实验方剂学杂志》2025年第9期124-133,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81960828,82160862);第五批全国中医临床优秀人才研修项目(国中医药人教函〔2022〕239号);首批陇原青年英才项目(中共甘肃省委人才工作领导小组〔2022〕5号)。

摘　　要：目的:观察黑逍遥散对阿尔茨海默病模型小鼠学习记忆能力的影响,并从核苷酸结合寡聚化结构域样受体3(NLRP3)/胱天蛋白酶-1(Caspase-1)/Gasdermin家族蛋白D(GSDMD)信号通路介导的神经炎症反应探讨其作用机制。方法:将SPF级4月龄APP/PS1小鼠随机分为模型组、MCC950组及黑逍遥散高、中、低剂量组,另将C57BL/6J小鼠作为空白组。适应性喂养7 d后,对各组小鼠分别进行干预,黑逍遥散高、中、低剂量组分别灌胃给药对应剂量药液(25.79、12.90、6.45 g·kg^(-1)·d^(-1)),MCC950组腹腔注射给药(10 mg·kg^(-1)·2 d^(-1)),空白组给予等体积生理盐水灌胃。干预90 d后用Y迷宫和Morris水迷宫试验测试学习记忆能力,苏木素-伊红(HE)染色观察海马神经元结构改变,免疫组化检测海马CA3区淀粉样蛋白前体(APP)表达水平,酶联免疫吸附测定法(ELISA)检测海马组织白细胞介素(IL)-10、IL-18、IL-1β表达水平,蛋白免疫印迹法(Western blot)检测海马组织NLRP3、Caspase-1、GSDMD、GSDMD-N蛋白表达,免疫荧光检测GSDMD-N和离子钙结合衔接分子-1(Iba-1)的共定位表达。结果:①Y迷宫实验结果显示,与空白组比较,模型组自发交替率显著减少(P<0.01);与模型组比较,黑逍遥散高、低剂量组自发交替率显著增加(P<0.01)。②Morris水迷宫实验结果显示,在1~4 d的定位航行测试中,随着训练时间的延长,各组逃避潜伏期时间均缩短。第4天,与空白组比较,模型组逃避潜伏期明显升高(P<0.05);与模型组比较,MCC950组和黑逍遥散低剂量组逃避潜伏期明显缩短(P<0.05)。在空间探索实验中,与空白组比较,模型组跨越平台位置的次数显著降低(P<0.01);与模型组比较,黑逍遥散低剂量组跨越平台次数明显增高(P<0.05)。③HE染色显示,模型组海马CA3区细胞损伤,排列松散且不规则、肿胀、边界不清,细胞核固缩、深染;MCC950与黑逍遥散各剂量组均能不同程度地改善APP/PS1海马CA3�Objective:To observe the effects of Hei Xiaoyaosan on the learning and memory abilities of Alzheimer's disease model mice(APP/PS1 mice),and to explore its mechanism through the inflammatory cascade mediated by nucleotidebinding oligomerization domain-like receptor family pyrin domain-containing 3(NLRP3)/cysteine aspartate-specific protease(Caspase-1)/gasdermin D(GSDMD)signaling pathway.Methods:SPF-grade 4-month-old APP/PS1 mice were randomly divided into the model group,MCC950 group,and Hei Xiaoyaosan high-,medium-,and low-dose groups.C57BL/6J mice were used as the blank group.After 7 days of adaptive feeding,mice in each group were intervened.The Hei Xiaoyaosan high-,medium-,and low-dose groups were given corresponding doses by gavage(25.79,12.90,6.45 g·kg^(-1)·d^(-1)),the MCC950 group was intraperitoneally injected with 10 mg·kg^(-1)·2 d^(-1),and the blank group received the same volume of physiological saline by gavage.After 90 days of intervention,the learning and memory abilities were assessed using the Y maze and Morris water maze tests.The structural changes of hippocampal neurons were observed by hematoxylin-eosin(HE)staining.The expression of amyloid precursor protein(APP)in the hippocampal CA3 region was detected by immunohistochemistry.Enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of interleukin(IL)-10,IL-18,and IL-1βin the hippocampus.Western blot was applied to detect the protein expression of NLRP3,Caspase-1,GSDMD,and GSDMD-N in the hippocampus.Immunofluorescence was used to detect the co-localization of GSDMD-N and ionized calcium-binding adapter molecule-1(Iba-1)in the hippocampus.Results:①In the Y maze test,compared with the blank group,the spontaneous alternation rate of the model group was significantly reduced(P<0.01).Compared with the model group,the spontaneous alternation rate in the Hei Xiaoyaosan high-and low-dose groups was significantly increased(P<0.01).②In the Morris water maze test,during the 1-4 days of the location navigation test,the escape latenc

关 键 词：阿尔茨海默病 黑逍遥散 神经炎症 核苷酸结合寡聚化结构域样受体3(NLRP3)/胱天蛋白酶-1(Caspase-1)/Gasdermin家族蛋白D(GSDMD)信号通路 实验研究 

分 类 号：R259[医药卫生—中西医结合] R587.1[医药卫生—中医内科学] R544.1[医药卫生—中医学]