枸杞多糖干预β-淀粉样蛋白1-42诱导SH-SY5Y细胞损伤:线粒体自噬的保护作用  

Lycium barbarum polysaccharide intervenes in SH-SY5Y cell injury induced by beta-amyloid protein 1-42: protective effect of mitochondrial autophagy

作　　者：苏琴 贾思玮 郭敏芳[2] 孟涛 李雁冰 穆秉桃 宋丽娟 马存根 尉杰忠[1,2,3] Su Qin;Jia Siwei;Guo Minfang;Meng Tao;Li Yanbing;Mu Bingtao;Song Lijuan;Ma Cungen;Yu Jiezhong(Neurobiology Research Center of Shanxi University of Chinese Medicine/Key Research Laboratory of State Administration of Traditional Chinese Medicine for Treatment of Multiple Sclerosis with Replenishing Qi and Huoxue Method,Jinzhong 030619,Shanxi Province,China;Institute of Brain Science of Shanxi Datong University/Molecular Cellular Immunology,Datong City Key Laboratory,Datong 037009,Shanxi Province,China;Datong Fifth People’s Hospital,Datong 037009,Shanxi Province,China)

机构地区：[1]山西中医药大学神经生物学研究中心/国家中医药管理局益气活血法治疗多发性硬化重点研究室,山西省晋中市030619 [2]山西大同大学脑科学研究所/分子细胞免疫学大同市重点实验室,山西省大同市037009 [3]大同市第五人民医院,山西省大同市037009

出　　处：《中国组织工程研究》2025年第31期6688-6696,共9页Chinese Journal of Tissue Engineering Research

基　　金：山西省基础研究计划项目(202303021211244),项目负责人:尉杰忠;山西省2022年度“四个一批”科技兴医创新计划项目(2022XM33),项目负责人:尉杰忠;山西省中医药科研课题计划项目(2023ZYYB2042),项目负责人:尉杰忠;国家中医药管理局科研课题(2023ZYYDA2038),项目负责人:马存根;山西中医药大学2022年度科技创新团队项目(2022TD2010),项目负责人:马存根;山西省卫健委2022年度中医药科研课题立项计划(2022ZYYC090),项目负责人:马存根;山西省基础研究计划项目(20210302123476),项目负责人:郭敏芳;大同市平台计划项目(2022082),项目负责人:尉杰忠;山西省卫健委科研课题计划项目(2021168),项目负责人:尉杰忠;山西省中医药管理局科研课题(2023ZYYB2042),项目负责人:尉杰忠;山西省中医药重点研究室(zyyyjs2024027),项目负责人:尉杰忠。

摘　　要：背景:神经退行性疾病与线粒体自噬调节失衡密切相关。课题组前期研究表明枸杞多糖具有神经保护作用,但其能否通过调控线粒体自噬来改善β-淀粉样蛋白1-42诱导的SH-SY5Y细胞损伤尚不明确。目的:探讨枸杞多糖对β-淀粉样蛋白1-42诱导的SH-SY5Y细胞损伤的保护作用及机制。方法:通过β-淀粉样蛋白1-42诱导SH-SY5Y细胞建立阿尔茨海默病细胞模型,并用枸杞多糖进行干预。将SH-SY5Y细胞分为3组:对照组,β-淀粉样蛋白1-42组(20μmol/Lβ-淀粉样蛋白1-42干预24 h),枸杞多糖组(先提前1 h加入1 g/L枸杞多糖形成保护作用,然后再加入20μmol/Lβ-淀粉样蛋白1-42与枸杞多糖共同干预24 h)。CCK-8法检测细胞活力;JC-1检测线粒体膜电位;TUNEL染色检测细胞凋亡;免疫荧光和Western blot检测突触、凋亡和线粒体自噬相关指标的表达。结果与结论:①与对照组比较,β-淀粉样蛋白1-42组细胞活力下降(P<0.05),细胞凋亡率上升(P<0.05),线粒体膜电位下降(P<0.05),促凋亡蛋白Bax、Caspase-3表达升高(P<0.05),抗凋亡蛋白Bcl-2表达降低(P<0.05),突触相关蛋白Syn、PSD-95表达降低(P<0.05),线粒体自噬相关蛋白Pink1、LC3A/B、Parkin、Beclin-1表达降低(P<0.05),P62表达升高(P<0.05);②与β-淀粉样蛋白1-42组比较,枸杞多糖组细胞活力升高(P<0.05),细胞凋亡率降低(P<0.05),线粒体膜电位上升(P<0.05),Bax和Caspase-3表达降低(P<0.05),Bcl-2表达升高(P<0.05),Syn和PSD-95表达升高(P<0.05),Pink1、LC3A/B、Parkin、Beclin-1表达升高(P<0.05),P62表达降低(P<0.05)。结果表明:枸杞多糖可能通过调控线粒体自噬来抑制β-淀粉样蛋白1-42诱导的SH-SY5Y细胞损伤,减少细胞凋亡,增加神经元突触可塑性。BACKGROUND:Neurodegenerative diseases are closely related to the imbalance of mitochondrial autophagy regulation.Previous studies by the research group have shown that lycium barbarum polysaccharide has neuroprotective effects,but whether it can improve the damage of SH-SY5Y cells induced byβ-amyloid protein 1-42 by regulating mitochondrial autophagy is still unclear.OBJECTIVE:To explore the protective effect and mechanism of Lycium barbarum polysaccharide on SH-SY5Y cells induced byβ-amyloid protein 1-42.METHODS:An Alzheimer's disease cell model was established by inducing SH-SY5Y cells withβ-amyloid protein 1-42,and then intervening with Lycium barbarum polysaccharide.SH-SY5Y cells were divided into three groups:control group,β-amyloid protein 1-42 group(20μmol/Lβ-amyloid protein 1-42 for 24 hours),and Lycium barbarum polysaccharide group(1 g/L Lycium barbarum polysaccharide was added 1 hour in advance to form a protective effect,and then 20μmol/Lβ-amyloid protein 1-42 was added to intervene with Lycium barbarum polysaccharide for 24 hours).CCK8 assay was used to detect cell viability.Mitochondrial membrane potential was detected by JC-1.TUNEL staining was used to detect cell apoptosis.Immunofluorescence and western blot assay were used to detect the expression of synaptic,apoptosis,and mitophagy-related indicators.RESULTS AND CONCLUSION:(1)Compared with the control group,the cell viability of theβ-amyloid protein 1-42 group decreased(P<0.05);cell apoptosis rate increased(P<0.05);mitochondrial membrane potential decreased(P<0.05);the expressions of pro-apoptotic proteins Bax and Caspase3 increased(P<0.05);the expression of anti-apoptotic protein Bcl-2 decreased(P<0.05);the expression levels of synaptic-related proteins Syn and PSD-95 decreased(P<0.05);the expression levels of mitochondrial autophagy-related proteins Pink1,LC3A/B,Parkin,and Beclin-1 decreased(P<0.05);and the expression of P62 increased(P<0.05).(2)Compared with theβ-amyloid protein 1-42 group,the cell viability in the Lycium barbarum po

关 键 词：枸杞多糖 阿尔茨海默病 SH-SY5Y细胞 线粒体自噬 Β-淀粉样蛋白 细胞凋亡 工程化细胞 

分 类 号：R459.9[医药卫生—治疗学] R392.12[医药卫生—临床医学] R277.7