原花青素对MPP+暴露的SH-SY5Y细胞线粒体质量控制系统的影响  

EFFECTS OF PROANTHOCYANIDINS ON MITOCHONDRIAL QUALITY CONTROL SYSTEM IN SH-SY5Y CELLS EXPOSED TO MPP+

作　　者：张秀文 张小强[1,2] 黄文迪 丁岚 张美荣 ZHANG Xiu-wen;ZHANG Xiao-qiang;HUANG Wen-di;DING Lan;ZHANG Mei-rong(School of Public Health,Southeast University,Nanjing 210009,China;Key Laboratory of Environmental Medicine Engineering of Ministry of Education,Southeast University,Nanjing 210009,China)

机构地区：[1]东南大学公共卫生学院,南京210009 [2]东南大学环境医学工程教育部重点实验室,南京210009

出　　处：《营养学报》2023年第2期172-180,共9页Acta Nutrimenta Sinica

基　　金：中央高校基本科研业务费专项资金资助和江苏省普通高校研究生科研创新计划资助项目(No.SJZZ16_0034)。

摘　　要：目的探究原花青素(proanthocyanidins,PAC)对N-甲基-4-苯基吡啶鎓碘化物(MPP+)诱导的SH-SY5Y细胞线粒体质量控制系统的影响。方法用MPP+处理SH-SY5Y细胞建立帕金森病细胞模型。将SH-SY5Y细胞随机分为对照组(control)、模型组(MPP+)、PAC低剂量组(MPP++1.0μg/mlPAC)、PAC中剂量组(MPP++5.0μg/mlPAC)和PAC高剂量组(MPP++10.0μg/mlPAC)。以MTT法检测细胞活力、ROS检测试剂盒检测ROS生成量、Westernblot法检测线粒体分裂、融合、生物发生、自噬相关蛋白的表达情况。结果与对照组比较,MPP+处理后SH-SY5Y细胞活力显著降低,活性氧(ROS)的积累显著增加。此外,与对照组相比,MPP+处理使线粒体分裂相关蛋白(FIS1和DRP1)表达水平显著升高、线粒体融合相关蛋白(MFN1、MFN2、OPA1)、线粒体生物发生相关蛋白(PGC-1α、TFAM、NRF1)、自噬标志相关蛋白(PINK1、Parkin、LC3Ⅱ/Ⅰ、PHB2)表达显著降低,P62蛋白表达显著增加。然而,PAC处理显著逆转了MPP+引起的这些改变,从而保护SH-SY5Y细胞免受MPP+诱导的神经毒性损伤,维持线粒体质量、数量及功能的动态平衡。结论:PAC通过改善线粒体氧化应激、动力学平衡以及自噬,缓解MPP+诱导的神经细胞毒性。[营养学报,2023,45(2):172-180]Objective To investigate the effects of proanthocyanidins(PAC)on mitochondrial quality control process in SH-SY5Y cells exposed to N-methyl-4-phenylpyridinium iodide(MPP+).Methods Cellular Parkinson's disease model was established by treating SH-SY5Y cells with MPP+.SH-SY5Y cells were randomly divided into the control group(Control),the model group(MPP+),the low dose group of PAC(MPP++1.0μg/ml PAC),the medium dose group of PAC(MPP++5.0μg/ml PAC)and the high dose group of PAC(MPP++10.0μg/ml PAC).The MTT method was used to detect cell viability,the reactive oxygen species(ROS)detection kit to detect ROS production,and the Western blot method to examine the expression of mitochondrial division,fusion,biogenesis,and autophagy-related proteins.Results Compared with the control group,the viability of SH-SY5Y cells was significantly reduced and the accumulation of ROS was significantly increased after MPP+treatment.In addition,MPP+treatment significantly increased the expression levels of mitochondrial division-related proteins(FIS1 and DRP1),and reduced the expression levels of mitochondrial fusion-related proteins(MFN1,MFN2,OPA1),and mitochondrial biogenesis-related proteins(PGC-1α,TFAM,NRF1),autophagy marker-related proteins(PINK1,Parkin,LC3II/Ⅰand PHB2)and P62 expression was also significantly increased.However,PAC treatment significantly reversed these changes induced by MPP+,and thus protected SH-SY5Y cells from MPP+-induced neurocytotoxic damage and maintained the dynamic balance of mitochondrial quality,quantity and function.Conclusion PAC alleviates MPP+-induced neurocytotoxicity by improving mitochondrial oxidative stress,kinetic homeostasis,and autophagy.[ACTA NUTRIMENTA SINICA,2023,45(2):172-180]

关 键 词：原花青素 帕金森病 线粒体 线粒体质量控制系统 

分 类 号：R151.41[医药卫生—营养与食品卫生学]