香草酸对朊病毒感染细胞PrP^(Sc)沉积抑制和细胞保护作用研究  

Inhibitory Effect on PrP^(Sc)Deposition and Cell Protection of Vanillic Acid in Prion⁃infected Cells

作　　者：王琳 杨微 姚强[7] 陈嘉 胡超 夏影 董小平[3,4,5] 石琦 陈志宝[1,2] 陈操 WANG Lin;YANG Wei;YAO Qiang;CHEN Jia;HU Chao;XIA Ying;DONG Xiaoping;SHI Qi;CHEN Zhibao;CHEN Cao(College of Coastal Agricultural Sciences,Guangdong Ocean University,Zhanjiang 524088,China;Shenzhen Research Institute,Guangdong Ocean University,Shenzhen 518108,China;National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;State Key Laboratory for Infectious Disease Control and Prevention,Beijing 102206,China;Center for Biosafety Mega-Science,Chinese Academy of Sciences,Wuhan 430071,China;National Health Commission Key Laboratory for Medical Virology and Viral Diseases,Beijing 102206,China;China Animal Disease Provention and Control Center,Beijing 100125,China)

机构地区：[1]广东海洋大学滨海农业学院,湛江524088 [2]广东海洋大学深圳研究院,深圳518108 [3]中国疾病预防控制中心病毒病预防控制所,北京102206 [4]传染病预防控制国家重点实验室,北京102206 [5]中国科学院生物安全大科学研究中心,武汉430071 [6]卫生部医学病毒和病毒病重点实验室,北京102206 [7]中国动物疫病预防控制中心,北京100125

出　　处：《病毒学报》2022年第4期896-904,共9页Chinese Journal of Virology

基　　金：传染病预防控制国家重点实验室青年基金(项目号:2021SKLID504),题目:朊病毒感染所致神经病理形成中宿主脑组织微环境天然免疫因子作用研究;传染病预防控制国家重点实验室优青申报资助课题(项目号:2019SKLID401),题目:宿主天然免疫在朊病毒清除和神经损伤中作用研究;国家自然科学基金面上项目(项目号:81772197),题目:朊病毒感染致脑组织微环境钙离子稳态变化及相关蛋白作用机制研究。

摘　　要：朊病毒病是一类致死性神经退行性疾病,目前无有效的治疗性药物和预防性疫苗。本研究使用香草酸处理朊病毒感染细胞的SMB‐S15细胞,分析其抑制PrP^(Sc)沉积和细胞保护作用。Western Blot结果显示,10μM的VA连续处理14 d后,SMB‐S15细胞中的PrP^(Sc)水平显著降低。同时,细胞内的活性氧和过氧化氢水平呈剂量依赖性降低,而HO‐1、GCLC和GCLM等抗氧化因子水平显著升高。细胞内的总谷胱甘肽和超氧化物歧化酶活性上调。VA连续处理14 d的SMB‐S15细胞中,UPR相关蛋白PERK、IRE1α、ATF6和GRP78表达上调,自噬系统活化。此外,VA处理可缓解朊病毒感染引起的SIRT3异常。本研究为朊病毒病治疗药物研发提供了新线索,同时VA对体外PrP^(Sc)的抑制作用及对朊病毒感染细胞的保护作用值得进一步研究。Prion diseases are types of fatal neurodegenerative diseases.Efficacious therapeutic drugs or prophylactic vaccines are not available.Inhibition of scrapie‐associated prion protein(PrP^(Sc))propagation and the cellular protectivity of vanillic acid(VA)on a prion‐infected cell line(SMB‐S15)were evaluated.Western blots showed that the PrP^(Sc)level in SMB‐S15 cells was reduced significantly after incubation with VA(10μM)for 14 days.Meanwhile,levels of reactive oxygen species and hydrogen peroxide were reduced in a dose‐dependent manner,whereas levels of some antioxidant factors,such as heme oxygenase‐1(HO‐1),glutamatecysteine ligase catalytic subunit,(GCLC)and glutamate‐cysteine ligase modifier subunit(GCLM),were increased significantly.Furthermore,the activities of total glutathione and superoxide dismutase were increased.VA‐treated SMB‐S15 cells showed up‐regulation of expression of UPR‐related proteins,including protein kinase R(PKR)‐like endoplasmic reticulum kinase(PERK),inositol‐requiring enzyme 1aα(IRE1α),activating transcription factor 6(ATF6),glucose‐regulated protein 78(GRP78),and activation of the autophagy system.The abnormalities in sirtuin‐3 expression caused by prion infection were relieved by VA treatment.Our study provides new clues for the development of drugs to treat prion diseases.PrP^(Sc)inhibition and the protective effects of VA on prion‐infected cells in vitro merit further study.

关 键 词：朊病毒 SMB‐S15细胞 香草酸 氧化应激 内质网应激 线粒体功能障碍 

分 类 号：R373.9[医药卫生—病原生物学]