摘要
目的探讨中药山茱萸鞣质对D-半乳糖(D-Galactose,D-gal)致阿尔茨海默病(Alzheimer爷s disease,AD)小鼠认知障碍的保护作用及药理机制。方法选取60只C57小鼠随机分为空白对照组、D-gal模型组、阳性对照组(维生素E 150 mg/kg)及山茱萸鞣质高、中、低(150、100、50 mg/kg)剂量组,每组10只。利用D-gal腹腔注射诱导AD小鼠模型,同时每天给予相应剂量的药物进行灌胃干预,实验周期维持8周。末次灌胃给药结束后,利用Morris水迷宫和旷场实验评估小鼠学习记忆与认知功能,使用分光光度法检测海马组织超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)、过氧化氢酶(catalase,CAT)抗氧化酶活力及丙二醛(malondialdehyde,MDA)含量,采用苏木精—伊红染色和Nissl染色观察海马区病理变化,并运用蛋白印迹法(Western blot,WB)考察突触相关蛋白微管关联蛋白-2(microtubule-associatedprotein-2,MAP-2)、突触后密度蛋白-95(postsynaptic density protein-95,PSD-95)、突触素(synaptophysin,SYN)及核因子E2相关因子2/抗氧化反应元件(nuclear factor E2-related factor 2/an-tioxidant response element,Nrf2/ARE)抗氧化信号通路相关蛋白Nrf2、血红素氧合酶-1(hemeoxygenase-1,HO-1)、醌氧化还原酶1(quinone oxidoreductase 1,NQO1)等表达变化水平。结果与D-gal模型组比较,山茱萸鞣质给药组小鼠在Morris水迷宫实验中的逃避潜伏期显著降低、目标区域停留时间和穿越平台次数显著增加(P<0.01);在旷场实验中的跨格次数、中心区域停留时间、平均速度及总路程均明显增加(P<0.01)。生化结果显示,与D-gal模型组比较,山茱萸鞣质给药组小鼠脑内SOD、GSH-Px及CAT等抗氧化酶活力显著上升(P<0.01),MDA含量明显下降(P<0.01)。苏木精—伊红染色与Nissl染色结果表明,与D-gal模型组比较,山茱萸鞣质给药组小鼠神经元受损情况明显减轻。Western blot结果显示,与D-gal模型组相比,山茱萸鞣质给药组小鼠脑内MAP-2、PSD-95、SYN、HO-1、NQO1、Nrf2等蛋白表达水平均显著增加(P<0.01)。结论山茱萸鞣质可改善D-gal致AD小鼠的认知障碍和神经元突触损伤,其潜在分子机制可能与上调Nrf2/ARE信号通路并减轻海马组织氧化应激有关。
Objective To explore the protective effect and molecular mechanism of tannin in the traditional Chinese medicine Cornus officinalis tannin(SZYT)on cognitive impairment in mice with Alzheimer's disease(AD)induced by D-galactose(D-gal).Methods Sixty C57 mice were randomly divided into the blank control group,the D-gal model group,the positive control group(vitamin E 150 mg/kg),and the high,medium,and low(150,100,and 50 mg/kg)dose groups of SZYT,with 10mice in each group.The AD mouse model was induced by intraperitoneal injection of D-gal,and the corresponding dose of the drug was administered by gavage intervention every day.The experimental period was maintained for 8 weeks.After the last gavage administration,the learning,memory and cognitive functions of mice were evaluated by Morris water maze and open field experiments.The activities of antioxidant enzymes such as superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),catalase(CAT)and the content of malondialdehyde(MDA)in hippocampal tissue were detected by spectrophotometry.The pathological changes in the hippocampus were observed by hematoxylin-eosin staining and Nissl staining.Western blot was used to investigate synaptic associated proteins microtubule-associated protein-2(MAP2),postsynaptic density protein-95(PSD-95),synaptophysin(SYN),nuclear factor E2-related factor 2/antioxidant response element(Nrf2/ARE),antioxidant signaling pathway related proteins Nrf2,heme oxygenase-1(HO-1),and quinone oxidoreductase 1(NQO1).Results Compared with the D-gal model group,the escape latency of mice in the SZYT group in the Morris water maze test was significantly reduced,and the stay time in the target area and the number of times they crossed the platform were significantly increased(P<0.01).In the open field experiment,the number of grid crossings,the duration of stay in the central area,the average speed and the total distance all increased significantly(P<0.01).The biochemical results showed that compared with the D-gal model group,the activities of antioxidant enzymes such as SOD,GSH-Px and CAT in the brains of mice in the SZYT administration group significantly increased(P<0.01),and the content of MDA significantly decreased(P<0.01).The results of HE and Nissl staining indicated that compared with the D-gal model group,the neuronal damage in the SZYT administration group of mice was significantly alleviated.The Western blot results showed that,compared with the D-gal model group,the expression levels of proteins such as MAP-2,PSD-95,SYN,HO-1,NQO1,and Nrf2 in the brain of mice in the SZYT administration group were significantly increased(P<0.01).Conclusion SZYT can improve cognitive impairment and neuronal synaptic injury in mice with AD induced by D-gal.The underlying molecular mechanism may be related to the upregulation of the Nrf2/ARE signaling pathway and the alleviation of oxidative stress in hippocampal tissue.
作者
陈鹏
陈小青
黎昌林
甘雄
刘刚
周本宏
CHEN Peng;CHEN Xiaoqing;LI Changlin;GAN Xiong;LIU Gang;ZHOU Benhong(Department of Pharmacy,Renmin Hospital of Wuhan University,Wuhan 430060,China)
出处
《环球中医药》
2026年第1期18-26,共9页
Global Traditional Chinese Medicine
基金
国家自然科学基金(82505051)
湖北省自然科学基金(2025AFB273)
湖北省中医药管理局中医药科研项目(ZY2025L034)
湖北省卫生健康科技项目(WJ2025M194)
湖北省教育厅科学研究计划指导性项目(B2024230)。
