基于肠道菌群探讨二精丸对AD模型大鼠认知功能的影响及机制  

Effect and Mechanism of Erjing Pills (二精丸) on Cognitive Function of AD Model Rats Based on Intestinal Flora

作　　者：黄丽萍[1,5] 卢龙辉 杨喜洋 谢永艳 徐子薇 朱徐东 王晶晶 黄梓轩 刘敏 吴毅 陈耀辉 HUANG Liping;LU Longhui;YANG Xiyang;XIE Yongyan;XU Ziwei;ZHU Xudong;WANG Jingjing;HUANG Zixuan;LIU Min;WU Yi;CHEN Yaohui(School of Pharmacy,Jiangxi University of Chinese Medicine,Nanchang 330004;Jiangxi Provincial People's Hospital,Nanchang 330006;The First Hospital of Putian City,Putian 351199;NMPA Key Laboratory of Quality Evaluation of Traditional Chinese Patent Medicine,Jiangxi Provincial Engineering Research Center for Drug and Medical Device Quality,Jiangxi Provincial Institute for Drug Control,Nanchang 330046;Key Laboratory of TCM Pharmacology of Jiangxi Province,Nanchang 330004;Nanchang Medical College,Nanchang 330052)

机构地区：[1]江西中医药大学药学院,南昌330004 [2]江西省人民医院,南昌330006 [3]莆田市第一医院,莆田351199 [4]国家药品监督管理局中成药质量评价重点实验室,江西省药品检验检测研究院,江西省药品与医疗器械质量工程研究中心,南昌330046 [5]江西省中药药理重点实验室,南昌330004 [6]南昌医学院,南昌330052

出　　处：《中药药理与临床》2023年第9期2-8,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：江西省主要学科学术和技术带头人资助项目(编号:20182BCB22005);江西省自然科学基金项目(编号:20171BAB205083);江西省重点研发计划项目(编号:20181BBC70046、20192BBG70071、20202BBGL73006);江西省中医药管理局课题(编号:2021A346);江西中医药大学校级科技创新团队发展计划(编号:CXTD22007);江西省教育厅重点项目(GJJ215601);江西省研究生创新专项资金项目(编号:YC2021-S505)。

摘　　要：目的:研究二精丸对阿尔茨海默病(AD)模型大鼠认知功能的影响,并从肠道菌群角度探讨其作用机制。方法:SD大鼠随机分为正常对照组、模型对照组、多奈哌齐1 mg/kg组、二精丸4.5、9.0 g/kg,每组14只。除正常对照组外,其余各组大鼠腹腔注射D-半乳糖100 mg/kg并双侧海马注射Aβ_(25-35),建立AD大鼠模型。连续灌胃给药5 w后,Morris水迷宫考察大鼠学习记忆能力。末次给药24 h后,取组织。16S rDNA高通量测序技术分析各组大鼠肠道菌群多样性和组成;HE染色观察大鼠结肠组织病理学改变;ELISA法检测大鼠海马组织炎症因子白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、IL-6含量;免疫组化法检测大鼠肠黏膜闭锁连接蛋白-1(zonula occludens-1,ZO-1)和紧密连接蛋白-1(Claudin-1)表达。结果:与正常对照组相比,模型对照组大鼠学习记忆能力显著降低,肠杆菌属数量显著增加,结肠组织损伤明显,病理学评分显著升高,海马组织IL-1β、TNF-α、IL-6含量显著升高,肠道ZO-1和Claudin-1蛋白阳性表达显著降低(P<0.01);与模型对照组比较,二精丸9 g/kg大鼠学习记忆能力显著提高,肠杆菌属数量显著减少(P<0.01),结肠组织形态明显改善,病理学评分显著降低(P<0.01),海马组织IL-1β、TNF-α、IL-6含量明显降低,肠道ZO-1和Claudin-1蛋白阳性表达显著升高(P<0.05或P<0.01)。结论:二精丸可能通过改善肠杆菌属的异常增殖,参与短链脂肪酸代谢,减少肠道炎症反应,恢复模型大鼠肠黏膜屏障功能,减少神经炎症因子水平,从而提高AD大鼠的认知功能。Objective:To study the effect of Erjing Pills(二精丸)on cognitive function in the Alzheimer's disease(AD)model in rats and explore its mechanism from the perspective of intestinal flora.Methods:SD rats were randomly divided into a normal control group,a model control group,a donepezil(1 mg/kg)group,and Erjing Pills(9.0 and 4.5 g/kg)groups,with 14 rats in each group.Except for the normal control group,rats in the other groups were injected with D-galactose(100 mg/kg)intraperitoneally,followed by bilateral hippocampal injection of Aβ25-35 to establish the AD model.After five weeks of continuous oral administration,the Morris water maze test was performed to assess the rats'learning and memory abilities.Tissue samples were collected 24 hours after the last administration.High-throughput sequencing of 16S rDNA was used to analyze the diversity and composition of intestinal flora in each group.Histopathological changes in the colon tissues were observed using HE staining.Immunohistochemistry was conducted to detect the expression of zonula occludens-1(ZO-1)and Claudin-1 in the rat intestinal mucosa.ELISA was performed to measure the levels of pro-inflammatory cytokines interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),and IL-6 in the rat hippocampus.Results:Compared with the normal control group,the model control group showed significantly impaired learning and memory abilities in rats,increased abundance of Enterobacter(P<0.01),obvious colonic tissue damage with significantly higher histopathological scores(P<0.01),decreased expression of intestinal ZO-1 and Claudin-1(P<0.01),and significantly elevated levels of IL-1β,TNF-α,and IL-6 in the hippocampus(P<0.01).Compared with the model control group,rats treated with Erjing Pills at 9.0 g/kg exhibited significantly improved learning and memory abilities(P<0.01),decreased abundance of Enterobacter(P<0.01),improved colonic tissue morphology with significantly lower histopathological scores(P<0.01),increased expression of intestinal ZO-1 and Claudin-1(P<

关 键 词：肠道菌群 二精丸 阿尔茨海默病 神经炎症 认知功能 

分 类 号：R285.5[医药卫生—中药学]