基于AMPK/PGC-1α/SIRT3通路研究不同剂型开心散改善线粒体功能防治认知障碍的作用机制  

Mechanism of Different Dosage Forms of Kaixinsan in Improving Mitochondrial Function for Prevention and Treatment of Cognitive Disorder Based on AMPK/PGC-1α/SIRT3 Pathway

作　　者：康舒悦 喻燕姿 孙家群 陈玟璇 杨雅琴[2] 王奇[1] 李伟荣[1] 姚丽梅[2] KANG Shuyue;YU Yanzi;SUN Jiaqun;CHEN Wenxuan;YANG Yaqin;WANG Qi;LI Weirong;YAO Limei(Science and Technology Innovation Center,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;College of Traditional Chinese Medicine and Health Care,Guangdong Food and Drug Vocational College,Guangzhou 510520,China)

机构地区：[1]广州中医药大学科技创新中心,广州510405 [2]广东食品药品职业学院中医保健学院,广州510520

出　　处：《中国实验方剂学杂志》2025年第7期15-24,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82074505,82374547);广东省基础与应用基础研究基金项目(2022A1515220121,2023A1515011835);广东省医学科学技术研究基金项目(B2023198)。

摘　　要：目的:研究不同剂型开心散(KXS)对阿尔茨海默病(AD)大鼠线粒体形态和功能的影响,并探讨其可能的作用机制。方法:雄性SD大鼠随机分为假手术组,模型组,KXS汤剂、散剂、配方颗粒剂组(3.08 g·kg^(-1)),多奈哌齐组(0.51 mg·kg^(-1)),每组各10只。采用双侧侧脑室注射链脲佐菌素(STZ)构建AD模型大鼠,连续给药30 d进行行为学检测,透射电镜观察线粒体形态,酶联免疫吸附测定法(ELISA)检测线粒体呼吸链复合物含量,JC-1染色法检测线粒体膜电位变化,生化试剂盒检测超氧化物歧化酶(SOD)活力、活性氧(ROS)含量,实时荧光定量聚合酶链式反应(Real-time PCR)检测腺苷酸活化蛋白激酶(AMPK)、过氧化物酶体增殖物激活受体γ辅激活因子-1α(PGC-1α)、沉默信息调节因子3(SIRT3)mRNA表达,蛋白免疫印迹法(Western blot)检测视神经萎缩1蛋白(OPA1)、线粒体分裂蛋白1(FIS1)、AMPK、磷酸化(p)-AMPK、PGC-1α、SIRT3蛋白表达。结果:与假手术组比较,模型组大鼠识别指数、自发交替反应率、逃避潜伏期、穿越平台次数、目标象限停留时间及目标象限路程占总路程百分比明显降低(P<0.05,P<0.01),海马组织线粒体损伤明显,线粒体呼吸链复合物含量显著减少(P<0.01)、线粒体膜电位明显降低(P<0.05),SOD活力降低、ROS含量升高(P<0.01),PGC-1α、SIRT3 mRNA表达显著减少(P<0.01),OPA1、p-AMPK/AMPK、PGC-1α、SIRT3蛋白表达降低,FIS1蛋白表达明显升高(P<0.05,P<0.01);与模型组比较,KXS各剂型组大鼠行为学指标明显改善(P<0.05,P<0.01),海马组织线粒体损伤减轻,内脊排列较清晰,线粒体呼吸链复合物含量明显增加(P<0.05,P<0.01),线粒体膜电位升高(P<0.05),SOD活力升高、ROS含量明显降低(P<0.05,P<0.01),PGC-1α、SIRT3 mRNA表达升高,OPA1、p-AMPK/AMPK、PGC-1α、SIRT3蛋白表达升高,FIS1蛋白表达明显降低(P<0.05,P<0.01);KXS各剂型组间比较,配方颗粒剂组大鼠自发交替反应率高于汤剂组Objective:To explore the effects of different dosage forms of Kaixinsan(KXS)on the morphology and function of mitochondria in rat models of Alzheimer's disease(AD)and potential mechanisms of action.Methods:Male SD rats were randomly assigned to a sham group,model group,treatment groups receiving KXS decoction,powders,and granules(3.08 g·kg^(-1)),as well as donepezil group(0.51×10^(-3)g·kg^(-1)),with 10 rats in each group.AD model was created using intracerebroventricular injection of streptozocin(STZ).After 30 days of administration,behavioral assessments were conducted,and mitochondrial morphology was observed using transmission electron microscopy.Mitochondrial respiratory chain complex content was measured via enzyme-linked immunosorbent assay(ELISA).Changes in mitochondrial membrane potential were measured via JC-1 staining,and superoxide dismutase(SOD)activity and reactive oxygen species(ROS)levels were measured via biochemical assays.The mRNA expression of adenosine 5'-monophosphate-activated protein kinase(AMPK),peroxisome proliferator-activated receptor gamma coactivator-1α(PGC-1α),and silent information regulator 3(SIRT3)was detected by realtime fluorescent quantitative polymerase chain reaction(Real-time PCR),and Western blot was used to examine the protein expression levels of optic atrophy protein1(OPA1),mitochondrial fission protein 1(FIS1),AMPK,p-AMPK,PGC-1α,and SIRT3.Results:Compared with the sham group,rats in the model group had significantly lower recognition index,spontaneous alternation rate,escape latency,number of platform crossings,time spent in the target quadrant,and percentage of distance traveled in the target quadrant distance(P<0.05,P<0.01).Significant mitochondrial damage was observed in the hippocampal tissue,with a marked decrease in mitochondrial respiratory chain complex content(P<0.01)and reduced mitochondrial membrane potential(P<0.05).Additionally,the SOD activity was reduced,while ROS levels were elevated(P<0.01).The mRNA expression of PGC-1αand SIRT3 was significantly

关 键 词：阿尔茨海默病 开心散 剂型 线粒体功能 腺苷酸活化蛋白激酶(AMPK)/过氧化物酶体增殖物激活受体γ辅激活因子-1α(PGC-1α)/沉默信息调节因子3(SIRT3) 

分 类 号：R289[医药卫生—方剂学] R285[医药卫生—中药学] R259[医药卫生—中医学]