中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (31): 6688-6696.doi: 10.12307/2025.545

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

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

苏  琴1，2，贾思玮1，2，郭敏芳2，孟  涛2，李雁冰1，2，穆秉桃2，宋丽娟1，马存根1，2，尉杰忠1，2，3   

  1. 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
  • 收稿日期:2024-06-27 接受日期:2024-08-31 出版日期:2025-11-08 发布日期:2025-02-24
  • 通讯作者: 尉杰忠，教授，博士生导师，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；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；大同市第五人民医院，山西省大同市 037009； 共同通讯作者：马存根，博士生导师，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；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
  • 作者简介:苏琴，女，湖北省石首市人，山西中医药大学在读硕士，主要从事神经变性疾病的基础研究。 共同第一作者：贾思玮，女，河南省郑州市人，山西中医药大学在读硕士，主要从事神经变性疾病的基础研究。
  • 基金资助:
    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

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

Su Qin1, 2, Jia Siwei1, 2, Guo Minfang2, Meng Tao2, Li Yanbing1, 2, Mu Bingtao2, Song Lijuan1, Ma Cungen1, 2, Yu Jiezhong1, 2, 3   

  1. 1Neurobiology 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; 2Institute of Brain Science of Shanxi Datong University/Molecular Cellular Immunology, Datong City Key Laboratory, Datong 037009, Shanxi Province, China; 3Datong Fifth People’s Hospital, Datong 037009, Shanxi Province, China
  • Received:2024-06-27 Accepted:2024-08-31 Online:2025-11-08 Published:2025-02-24
  • Contact: Yu Jiezhong, Professor, Doctoral supervisor, 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; Co-corresponding author: Ma Cungen, Doctoral supervisor, 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
  • About author:Su Qin, Master candidate, 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; Jia Siwei, Master candidate, 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 Su Qin and Jia Siwei contributed equally to this article.
  • Supported by:
    Shanxi Basic Research Program, No. 202303021211244 (to YJZ); Shanxi 2022 "Four Batches" Science and Technology Innovation Program, No. 2022XM33 (to YJZ); Shanxi Traditional Chinese Medicine Research Project, No. 2023ZYYB2042 (to YJZ); State Administration of Traditional Chinese Medicine Research Project, No. 2023ZYYDA2038 (to MCG); 2022 Science and Technology Innovation Team Project of Shanxi University of Chinese Medicine, No. 2022TD2010 (to MCG); The 2022 TCM Research Project Plan of Shanxi Provincial Health Commission, No. 2022ZYYC090 (to MCG); Basic Research Plan Project of Shanxi Province, No. 20210302123476 (to GMF); Platform Plan Project of Datong City, No. 2022082 (to YJZ); Research Project Plan Project of Shanxi Provincial Health Commission, No. 2021168 (to YJZ); Shanxi Provincial Administration of Traditional Chinese Medicine Research Project, No. 2023ZYYB2042 (to YJZ); Shanxi Provincial Key Research Office of Traditional Chinese Medicine, No. zyyyjs2024027 (to YJZ)

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摘要：

文题释义：

枸杞多糖：是从枸杞中提取而得的一种水溶性多糖，是枸杞中的主要功效成分，具有抗氧化活性、抗肿瘤特性、免疫调节和神经保护等多种作用。 
线粒体自噬：是一种细胞自噬的重要形式，主要通过去除损伤或功能失调的线粒体来维持细胞内线粒体的质量和功能。

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(20 μmol/L β-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20 μmol/L β-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。

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

Abstract: 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 by β-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 by β-amyloid protein 1-42.  
METHODS: An Alzheimer's disease cell model was established by inducing SH-SY5Y cells with β-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 (20 μmol/L β-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 20 μmol/L β-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 the β-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 the β-amyloid protein 1-42 group, the cell viability in the Lycium barbarum polysaccharide group was increased (P < 0.05); the apoptosis rate was decreased (P < 0.05); the mitochondrial membrane potential was increased (P < 0.05); the expression levels of Bax and Caspase3 were decreased (P < 0.05); the expression of Bcl-2 was increased (P < 0.05); the expressions of Syn and PSD-95 were increased (P < 0.05); the expression levels of Pink1, LC3A/B, Parkin, and Beclin-1 were increased (P < 0.05), and the expression of P62 was decreased (P < 0.05). These findings indicate that Lycium barbarum polysaccharide may inhibit β-amyloid protein 1-42-induced damage to SH-SY5Y cells by regulating mitophagy, reduce cell apoptosis, and increase neuronal synaptic plasticity. 


Key words: lycium barbarum polysaccharide, Alzheimer’s disease, SH-SY5Y cell, mitophagy, β-amyloid protein, apoptosis, engineered cells

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