丹皮酚对人神经母细胞瘤细胞帕金森病模型的保护作用及其初步机制  

Protective effects of paeonol on human neuroblastoma cells in Parkinson's disease model and its underlying mechanism

作　　者：孙胜男 和璐璐 秦劭晨 徐磊 王利然 于保锋 马存根 樊慧杰 柴智 Sun Sheng-Nan;He Lu-Lu;Qin Shao-Chen;Xu Lei;Wang Li-Ran;Yu Bao-Feng;Ma Cun-Gen;Fan Hui-Jie;Chai Zhi(The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine/Neurobiology Research Center,Shanxi University of Chinese Medicine,Jinzhong,Shanxi 030619,China;Biochemistry and Molecular Biology Teaching and Research Office,School of Basic Medicine,Shanxi Medical University,Taiyuan,Shanxi 030001,China;The First Clinical College of Shanxi University of Chinese Medicine,Taiyuan,Shanxi 030024,China;The Second Clinical College of Shanxi University of Chinese Medicine,Taiyuan,Shanxi 030012,China)

机构地区：[1]山西中医药大学多发性硬化益气活血重点研究室/神经生物学研究中心,山西晋中030619 [2]山西医科大学基础医学院生物化学与分子生物学教研室,山西太原030001 [3]山西中医药大学第一临床学院,山西太原030024 [4]山西中医药大学第二临床学院,山西太原030012

出　　处：《解放军医学杂志》2025年第1期69-75,共7页Medical Journal of Chinese People's Liberation Army

基　　金：国家中医药管理局青年岐黄学者培养项目[国中医药人教函(2022)256号];山西省重点国别科技合作项目(202204041101002);山西省青年拔尖人才支持计划项目(晋组办字[2019]35号文);山西省省筹资金资助回国留学人员科研项目(2021-142)。

摘　　要：目的探究丹皮酚(PAE)对过表达α-突触核蛋白(α-Syn)诱导的人神经母细胞瘤细胞(SH-SY5Y)损伤中自噬的影响及其相关作用机制。方法以SH-SY5Y细胞为对照组,含A53T-α-Syn突变的SH-SY5Y细胞为模型组、PAE(150μg/ml)组、3-甲基腺嘌呤(3-MA,1 mmol/L)组和PAE(150μg/ml)+3-MA(1 mmol/L)组。采用CCK-8法检测各组细胞存活率,光学显微镜下观察细胞形态,Western blotting检测α-Syn和自噬通路相关蛋白Beclin-1、p62、微管相关蛋白轻链3-Ⅱ(LC3-Ⅱ)、磷酸化c-Jun氨基末端激酶(p-JNK)、Bcl-2表达水平。结果与对照组比较,模型组细胞存活率明显降低(P<0.01),α-Syn蛋白表达水平明显增高(P<0.001),自噬相关蛋白LC3-Ⅱ、Beclin-1蛋白表达水平明显降低(P<0.01或P<0.05),自噬底物蛋白p62蛋白表达水平升高(P<0.05),调节自噬通路相关蛋白p-JNK、Bcl-2蛋白表达水平降低(P<0.05或P<0.01)。与模型组比较,PAE组细胞存活率增高(P<0.01),α-Syn、p62蛋白表达水平降低(P<0.01或P<0.05),LC3-Ⅱ、Beclin-1、p-JNK、Bcl-2蛋白表达水平增高(P<0.05)。与PAE组比较,PAE+3-MA组细胞α-Syn蛋白表达水平增高(P<0.05)。结论PAE可缓解A53T-α-Syn突变诱导的SH-SY5Y细胞损伤,并通过激活自噬途径清除过表达的α-Syn,其机制可能与上调JNK/Bcl-2介导的自噬途径有关。Objective To investigate the protective effects of paeonol(PAE)on autophagy in human neuroblastoma cells(SH-SY5Y)induced by overexpression ofα-synuclein(α-Syn),and to explore its related mechanism.Methods SH-SY5Y cells served as control group,while those induced with A53T-α-Syn mutation were used as model group.Additional groups included PAE(150μg/ml)group,3-MA(1 mmol/L)group,and PAE(150μg/ml)+3-MA(1 mmol/L)group.Cell viability was assessed using CCK-8 method,cell morphology was observed under an optical microscope,and protein expressions ofα-Syn,LC3-Ⅱ,p62,Beclin-1,phosphorylated c-Jun N-terminal kinase(p-JNK),and p-Bcl-2 were determined by Western blotting.Results Compared with control group,model control exhibited decreased cell survival(P<0.01),increasedα-Syn expression(P<0.001),reduced expression of autophagy-related proteins LC3-Ⅱand Beclin-1(P<0.01,P<0.05),elevated autophagy substrate protein p62(P<0.05),and decreased expression of autophagy pathway-related proteins p-JNK and Bcl-2(P<0.05,P<0.01).Compared with model group,PAE group showed increased cell survival(P<0.01),decreasedα‐Syn and p62 protein expression(P<0.01,P<0.05),and increased expression of LC3-Ⅱ,Beclin-1,p-JNK and Bcl-2(P<0.05).Compared with PAE group,3-MA+PAE group demonstrated increasedα-Syn expression(P<0.05).Conclusions PAE could attenuate the injury of SH-SY5Y cells induced by A53T-α-Syn and eliminate over-expressedα‐Syn by activating autophagy pathway,which may be associated with the upregulation of JNK/Bcl-2 mediated autophagy pathway.

关 键 词：丹皮酚 帕金森病 Α-突触核蛋白 自噬 

分 类 号：R742.5[医药卫生—神经病学与精神病学]