基于PINK1/Parkin信号通路探讨黄芪散改善阿尔茨海默病大鼠海马突触可塑性的机制  

Mechanism of Huangqisan improves hippocampal synaptic plasticity in Alzheimer′s disease rats based on PINK1/Parkin signaling pathway

作　　者：张运辉 袁云川 杨梦琳 伍大华[2,3] 刘霞 杨昆[1] 程妍 ZHANG Yun-hui;YUAN Yun-chuan;YANG Meng-lin;WU Da-hua;LIU Xia;YANG Kun;CHENG Yan(Chongqing Three Gorges Medical College,Chongqing 404120,China;Hunan University of Chinese Medicine,Changsha 410208,China;Affiliated Hospital of Hunan Provincial Academy of Traditional Chinese Medicine,Changsha 410006,China)

机构地区：[1]重庆三峡医药高等专科学校,重庆404120 [2]湖南中医药大学,长沙410208 [3]湖南省中医药研究院附属医院,长沙410006

出　　处：《天然产物研究与开发》2025年第4期612-623,共12页Natural Product Research and Development

基　　金：国家自然科学基金(81874462);万州区博士直通车项目(wzst20230415);重庆市中医药传承创新团队-三峡库区道地药材保护与利用多学科交叉创新团队(CXTD202303);重庆市自然科学基金面上项目(CSTB2024NSCQ-MSX0040)。

摘　　要：本研究旨在探究黄芪散对阿尔茨海默病(Alzheimer′s disease,AD)大鼠模型中突触可塑性的影响,并分析其潜在机制。SPF级雄性SD大鼠50只,随机分为正常组(normal group,Norm)、模型组(model group,Mod)、黄芪散低剂量组(Huangqisan low dose group,HQS-L;1.2 g/kg)、黄芪散高剂量组(Huangqisan high dose group,HQS-H;4.8 g/kg)及阳性对照组(positive group,Pos),每组10只。除正常组外,其余组双侧海马注射β淀粉样蛋白25-35造模。采用Morris水迷宫实验检测大鼠学习记忆功能;Nissl染色检测大鼠海马组织神经元损伤情况;JC-1探针流式细胞术检测海马组织线粒体膜电位水平;透射电镜观察大鼠海马区神经元和突触超微结构;ELISA法检测大鼠海马组织炎症因子白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平;流式细胞术检测海马活性氧(reactive oxygen species,ROS)水平;蛋白免疫印迹(Western blot,WB)法检测大鼠海马中PTEN诱导激酶1(PTEN-induced kinase 1,PINK1)、帕金蛋白(Parkin E3 ubiquitin protein ligase,Parkin)、微管相关蛋白1A/1B-轻链3(microtuble-associated protein light chain 3,LC3B)、泛素结合蛋白p62(ubiquitin-binding protein p62,p62)、动力相关蛋白1(dynmin-related protein 1,Drp1)、线粒体融合素1(mitofusion 1,Mfn1)、线粒体融合素2(mitofusion 1,Mfn2)、Nod样受体蛋白3(NOD-like receptor protein 3,NLRP3)、生长相关蛋白43(growth-associated protein 43,GAP43)、N-甲基-D-天冬氨酸受体2B亚基(N-methyl-D-aspartate receptor(NMDAR)subunit 2B,NR2B)、突触后致密蛋白95(postsynaptic dense protein95,PSD95)、突触素(synaptosin,SYP)、脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)蛋白表达。与Norm组比较,Mod组大鼠学习记忆功能显著降低(P<0.01),海马组织神经元胞内尼氏体减少或消失,海马区的神经元和突触超微结构出现明显的病理改变,线粒体膜电位明显This study aims to explore the impact of Huangqisan on synaptic plasticity in rat model of Alzheimer′s disease and to analyzed its potential mechanism.Fifty SPF grade male SD rats were randomly divided into normal group(Norm),model group(Mod),Huangqisan low dose group(HQS-L,1.2 g/kg),Huangqisan high dose group(HQS-H,4.8 g/kg)and positive group(Pos),with 10 rats in each group.Except for the normal group,the rats in other groups were injected withβ-amyloid 25-35 into bilateral hippocampal to establish the model.Morris water maze test was used to detect the learning and memory function of the rats.Nissl staining was used to detect neuronal damage in hippocampus of rats.JC-1 probe was applied to detect the mitochondrial membrane potential in hippocampus by flow cytometry.Transmission electron microscope was used to detect the ultrastructural observation of neurons and synapses in rat hippocampus.ELISA was performed to measure the levels of pro-inflammatory cytokines interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)in the rat hippocampus.Flow cytometry was used to detect the level of hippocampus reactive oxygen species(ROS).The expressions of PTEN-induced kinase 1(PINK1),Parkin E3 ubiquitin protein ligase(Parkin),microtuble-associated protein light chain 3(LC3B),ubiquitin-binding protein p62(p62),dynamin-related protein(Drp1),mitofusion 1(Mfn1),mitofusion 2(Mfn2),NOD-like receptor protein 3(NLRP3),growth-associated protein 43(GAP43),N-methyl-D-aspartate receptor(NMDAR)subunit 2B(NR2B),postsynaptic dense protein 95(PSD95),synaptosin(SYP)and brain-derived neurotrophic factor(BDNF)proteins in the hippocampus were detected by Western blot(WB).Compared with Norm group,the learning and memory function of rats in the Mod group was significantly reduced(P<0.01).Nissl bodies in neurons of hippocampus decreased or disappeared.The ultrastructure of neurons and synapses in hippocampus presented obvious pathological changes.Mitochondrial membrane potential decreased significantly and the structure

关 键 词：黄芪散 阿尔茨海默病 线粒体自噬 PTEN诱导激酶1/帕金蛋白信号通路 NLRP3炎症小体 

分 类 号：R285.5[医药卫生—中药学]