文献类型：期刊文献

中文题名：基于ATP/P2X7R/NF-κB信号通路探讨黑逍遥散对APP/PS1小鼠神经炎症的影响

英文题名：Effects of Heixiaoyao Powder on neuroinflammation in APP/PS1 mice via ATP/P2X7R/NF-κB signaling pathway

作者：孟志鹏[1];吕育洁[1];胡韵韵[1];杨娇[1];陈怡琴[1];王虎平[1,2]

第一作者：孟志鹏

机构：[1]甘肃中医药大学,甘肃兰州730000;[2]甘肃省中医方药挖掘与创新转化重点实验室,甘肃省中药新产品创制工程实验室,甘肃兰州730000

第一机构：甘肃中医药大学

年份：2025

卷号：47

期号：1

起止页码：51

中文期刊名：中成药

外文期刊名：Chinese Traditional Patent Medicine

收录：;北大核心:【北大核心2023】；

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项目(国中医药人教函[2022]239号);甘肃省自然科学基金项目(22JR11RA113);首批陇原青年英才项目(中共甘肃省委人才工作小组[2022]25号)。

语种：中文

中文关键词：黑逍遥散;阿尔茨海默病;神经炎症;ATP/P2X7R/NF-κB信号通路;β-淀粉样蛋白

外文关键词：Heixiaoyao Powder;Alzheimer’s disease;neuroinflammation;ATP/P2X7R/NF-κB signaling pathway;amyloid proteinβ

摘要：目的探究黑逍遥散对APP/PS1双转基因小鼠神经炎症的影响。方法16周龄雄性APP/PS1双转基因小鼠随机分为模型组、BBG组(P2X7R特异性拮抗剂,30 mg/kg)及黑逍遥散高、中、低剂量组(22.10、11.05、5.53 g/kg),以同周龄、同系种雄性C57BL/6J小鼠为空白组,每组12只。空白组和模型组灌胃生理盐水,其余各组灌胃相应剂量药物,给药90 d后,Morris水迷宫实验检测学习记忆能力,HE染色观察海马组织病理变化,免疫荧光染色检测海马组织MyD88表达,ELISA法检测海马组织促炎因子(TNF-α、IL-6)、抗炎因子(IL-10)及ATP、β-淀粉样蛋白(Aβ)水平,RT-qPCR法检测海马组织P2X7R、TLR4、MyD88、NF-κB-P 65 mRNA表达,Western blot法检测海马组织P2X7R、TLR4、MyD88、NF-κB-P65、p-NF-κB-P65蛋白表达。结果与空白组比较,模型组小鼠逃避潜伏期延长(P<0.01),穿越平台次数减少(P<0.01);海马神经元细胞数量减少,排列不规则,细胞质染色加深;海马组织MyD88免疫荧光表达升高(P<0.01),IL-10水平降低(P<0.01),TNF-α、IL-6、ATP、Aβ水平升高(P<0.01),P2X7R、TLR4、MyD88 mRNA及蛋白表达升高(P<0.01),p-NF-κB-P65蛋白表达升高(P<0.01)。与模型组比较,黑逍遥散各剂量组及BBG组小鼠逃避潜伏期缩短(P<0.01),穿越平台次数增加(P<0.01);海马神经元细胞数量增加,排列较为整齐;海马组织IL-10水平升高(P<0.01),TNF-α、IL-6、ATP、Aβ水平降低(P<0.05,P<0.01),P2X7R、TLR4、MyD88 mRNA及蛋白表达降低(P<0.05,P<0.01),p-NF-κB-P65蛋白表达降低(P<0.05,P<0.01);黑逍遥散高、中剂量组及BBG组MyD88免疫荧光表达降低(P<0.05,P<0.01)。结论黑逍遥散能显著改善APP/PS1模型小鼠学习记忆能力,其机制可能与降低脑内Aβ异常聚集、抑制ATP/P2X7R/NF-κB信号通路活化、减轻脑内神经炎症有关。
AIM To investigate the effects of Heixiaoyao Powder on neuroinflammation in APP/PS1 transgenic mice.METHODS The 16-week-old male APP/PS1 transgenic mice were randomly divided into the model group,the BBG group(P2X7R specific antagonist,30 mg/kg)and high,medium and low dose Heixiaoyao Powder groups of(22.10,11.05,5.53 g/kg),in contrast to the male C57BL/6J mice of the same age and the same strain of the blank groups,with 12 mice in each group.When normal saline by gavage was dosed upon the blank group and the model group,and the other groups were treated with corresponding drug by gavage.After 90 days of administration,the mice had their learning and memory ability detected by Morris water maze test;their hippocampal pathological changes observed with HE staining;their MyD88 expression detected by immunofluorescence staining;their hippocampal levels of pro-inflammatory factors(TNF-α,IL-6),anti-inflammatory factors(IL-10),ATP and amyloid proteinβ(Aβ)detected by ELISA;their hippocampal mRNA expressions of P2X7R,TLR4,MyD 88 and NF-κB-P 65 detected by RT-qPCR method;and their hippocampal protein expressions of P2X7R,TLR4,MyD88,NF-κB-P65 and p-NF-κB-P65 detected by Western blot.RESULTS Compared with the blank group,the model group demonstrated prolonged escape latency and reduced frequency in crossing platform(P<0.01);decreased number of hippocampal neurons,deranged neurons,and darker cytoplasm staining;increased immunofluorescence expression of hippocampal MyD88(P<0.01);decreased IL-10 level(P<0.01);increased levels of TNF-α,IL-6,ATP and Aβ(P<0.01);increased mRNA and protein expressions of P2X7R,TLR4 and MyD88(P<0.01),and increased protein expression of p-NF-κB-P65(P<0.01).Compared with the model group,the groups intervened with Heixiaoyao Powder or BBG demonstrated shortened escape latency and increased frequency in crossing platform(P<0.01);more number of neatly arranged hippocampal neurons;increased hippocampal IL-10 level(P<0.01),decreased levels of TNF-α,IL-6,ATP and Aβ(P<0.05,P<0.01);decreased mRNA and protein expressions of P2X7R,TLR4 and MyD88(P<0.05,P<0.01);and decreased protein expression of p-NF-κB-P65(P<0.05,P<0.01).Except the low dose Heixiaoyao Powder group,the other treatment groups shared decreased immunofluorescence expression of MyD88(P<0.05,P<0.01).CONCLUSION Heixiaoyao Powder can significantly improve the learning and memory ability of APP/PS1 model mice,and its mechanism may lie in its function in alleviating cerebral neuroinflammation by reducing the abnormal Aβaggregation via inhibiting the activation of ATP/P2X7R/NF-κB signaling pathway.