基于LKB1/AMPK/mTOR通路调节自噬探讨固本健脑液对阿尔茨海默病的作用机制  

Mechanism of Guben Jiannao Liquid on Alzheimer's disease by regulating autophagy based on LKB1/AMPK/mTOR pathway

作　　者：张婧凡 龙清华 曾楚华 陈益敏 谢喆尧 蔡元钦 王曦 ZHANG Jing-fan;LONG Qing-hua;ZENG Chu-hua;CHEN Yi-min;XIE Zhe-yao;CAI Yuan-qin;WANG Xi(Medical School,Hubei Minzu University,Enshi 445000,China;School of Basic Medical Sciences,Yunnan University of Chinese Medicine,Kunming 650500,China;Hubei Provincial Key Laboratory of Occurrence and Intervention of Rheumatic Diseases,Hubei Minzu University,Enshi 445000,China;Minda Hospital of Hubei Minzu University,Enshi 445000,China)

机构地区：[1]湖北民族大学医学部,湖北恩施445000 [2]云南中医药大学基础医学院,云南昆明650500 [3]湖北民族大学风湿性疾病发生与干预湖北省重点实验室,湖北恩施445000 [4]湖北民族大学附属民大医院,湖北恩施445000

出　　处：《中国中药杂志》2025年第2期293-300,共8页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金地区基金项目(82060831)。

摘　　要：基于肝激酶B1(liver kinase B1,LKB1)/腺苷酸活化蛋白激酶(adenosine monophosphate-activated protein kinase,AMPK)/哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)通路调节自噬探讨固本健脑液对阿尔茨海默病(Alzheimer's disease,AD)的作用机制。雄性SD大鼠随机分为空白组、模型组、固本健脑液低剂量组、固本健脑液高剂量组和雷帕霉素组,每组10只。除空白组外,其余组大鼠均双侧海马注射β-淀粉样蛋白(β-amyloid,Aβ)1-42建立AD模型。固本健脑液低剂量组(6.21 g·kg^(-1))、固本健脑液高剂量组(12.42 g·kg^(-1))、雷帕霉素组(1 mg·kg^(-1))给予相应药物灌胃,空白组、模型组给予等体积生理盐水灌胃,持续4周。Morris水迷宫检测各组大鼠学习记忆能力;苏木精-伊红(hematoxylin-eosin,HE)染色、尼氏染色观察大鼠海马CA1区神经元及尼氏体形态数量变化;免疫组化检测大鼠海马CA1区Aβ阳性细胞表达;透射电镜观察大鼠海马组织超微结构变化;Western blot检测大鼠海马组织LKB1、p-AMPK/AMPK、p-mTOR/mTOR、Beclin1、p62、LC3-Ⅱ的蛋白表达水平。结果显示,与空白组比较,模型组大鼠逃避潜伏期升高,穿越平台次数、在平台象限停留时间减少;海马区神经元数量减少、形态受损,Aβ阳性细胞表达升高;LKB1、p-AMPK/AMPK、Beclin1、LC3-Ⅱ表达水平降低,p-mTOR/mTOR、p62表达水平升高。与模型组比较,固本健脑液低、高剂量组大鼠逃避潜伏期降低,穿越平台次数、在平台象限停留时间升高,神经元数量增多,Aβ阳性细胞表达减少,海马组织自噬溶酶体数量增加;固本健脑液低剂量组大鼠海马LKB1、Beclin1、LC3-Ⅱ表达水平升高;固本健脑液高剂量组大鼠海马组织LKB1、p-AMPK/AMPK、Beclin1、LC3-Ⅱ表达水平升高,p-mTOR/mTOR、p62表达水平降低。以上结果显示,固本健脑液可以改善AD大鼠认知障碍,其作用机制可能与激活LKB1/AMPK/mTOR信号通路,上调自噬水平�This study explores the mechanism of Guben Jiannao Liquid on Alzheimer's disease(AD)by regulating autophagy based on the liver kinase B1(LKB1)/adenosine monophosphate-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)pathway.Male SD rats were randomly divided into the blank group,model group,low-dose and high-dose groups of Guben Jiannao Liquid,and rapamycin group,with 10 rats in each group.Except for the blank group,all other groups of rats were injected bilaterally in the hippocampus withβ-amyloid(Aβ)1-42 to establish the AD model.The low-dose(6.21 g·kg^(-1))and high-dose(12.42 g·kg^(-1))groups of Guben Jiannao Liquid and rapamycin group(1 mg·kg^(-1))were given the corresponding drugs by gavage,and the blank and model groups were given an equal volume of saline by gavage for four weeks.Morris water maze was used to test the learning and memory ability of rats in each group;hematoxylin-eosin(HE)and Nissl staining were used to observe the morphological and quantitative changes of neurons and Nissl bodies in the CA1 region of rat hippocampus;immunohistochemistry was utilized to detect Aβ-positive cell expression in the CA1 region of rat hippocampus;transmission electron microscopy was employed to observe ultrastructural changes in rat hippocampal tissue,and Western blot was used to examine the protein expression levels of LKB1,p-AMPK/AMPK,p-mTOR/mTOR,Beclin1,p62,and LC3-Ⅱin the hippocampal tissue of the rats.The results showed that compared with those in the blank group,rats in the model group had elevated evasion latency and decreased number of platform transversal and residence time in the platform quadrant.The number of neurons in the hippocampal area was reduced,and the morphology was impaired.The average integral optical density value of Aβ-positive cells was elevated;the expression levels of LKB1,p-AMPK/AMPK,Beclin1,and LC3-Ⅱwere decreased,and the expression levels of p-mTOR/mTOR and p62 were increased.Compared with those in the model group,rats in the low-dose and high-dose groups

关 键 词：阿尔茨海默病 固本健脑液 自噬 LKB1/AMPK/mTOR信号通路 

分 类 号：R73[医药卫生—肿瘤]