Recent increases in infectious diseases affecting the central nervous system have raised concerns about their role in neuroinflammation and neurodegeneration.Viral pathogens or their products can invade the central ne...Recent increases in infectious diseases affecting the central nervous system have raised concerns about their role in neuroinflammation and neurodegeneration.Viral pathogens or their products can invade the central nervous system and cause damage,leading to meningitis,encephalitis,meningoencephalitis,myelitis,or post-infectious demyelinating diseases.Although neuroinflammation initially has a protective function,chronic inflammation can contribute to the development of neurodegenerative diseases.Mechanisms such as protein aggregation and cellular disturbances are implicated with specific viruses such as herpes simplex virus type 1 and Epstein-Barr virus being associated with Alzheimer's disease and multiple sclerosis,respectively.Extracellular nucleotides,particularly adenosine triphosphate and its metabolites are released from activated,infected,and dying cells,acting as alarmins mediating neuroinflammation and neurodegeneration.When viruses infect central nervous system cells,adenosine triphosphate is released as an alarmin,triggering inflammatory responses.This process is mediated by purinergic receptors,divided into two families:P1,which responds to adenosine,and P2,activated by adenosine triphosphate and other nucleotides.This review highlights how specific viruses,such as human immunodeficiency virus type 1,Theiler's murine encephalomyelitis virus,herpes simplex virus type 1,Epstein-Barr virus,dengue virus,Zika virus,and severe acute respiratory syndrome coronavirus 2,can initiate inflammatory responses through the release of extracellular nucleotides,particularly adenosine triphosphate,which act as critical mediators in the progression of neuroinflammation and neurodegenerative disorders.A better understanding of purinergic signaling pathways in these diseases may suggest new potential therapeutic strategies for targeting neuroinflammation to mitigate the long-term consequences of viral infections in the central nervous system.展开更多
目的:探究多聚胞嘧啶结合蛋白2[poly(C)-binding protein 2,PCBP2]如何通过调节铁死亡参与大别班达病毒(Dabie Banda virus,DBV)感染后的致病过程及其作用机制。方法:以人单核细胞系THP-1为模型,采用qRT-PCR和Western blot技术检测DBV...目的:探究多聚胞嘧啶结合蛋白2[poly(C)-binding protein 2,PCBP2]如何通过调节铁死亡参与大别班达病毒(Dabie Banda virus,DBV)感染后的致病过程及其作用机制。方法:以人单核细胞系THP-1为模型,采用qRT-PCR和Western blot技术检测DBV感染的THP-1细胞中PCBP2的mRNA及蛋白表达水平。通过透射电镜观察病毒感染下的线粒体结构变化,在THP-1细胞中构建了慢病毒介导的PCBP2过表达和敲低稳转细胞系。FerroOrange荧光探针检测Fe^(2+)水平,2,7-二氯荧光素二乙酸酯(2,7-dichlorofluorescein diacetate,DCFH-DA)探针测定活性氧(reactive oxygen species,ROS)水平,Western blot检测铁死亡相关溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)和谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)蛋白表达,以评估PCBP2调控对铁死亡的影响。使用铁死亡诱导剂(RSL3、erastin)和抑制剂(Fer-1、Lip-1)处理细胞,qRT-PCR和免疫荧光检测病毒复制水平变化,探索PCBP2是否可以通过调控铁死亡影响DBV复制。结果:在DBV感染的细胞模型中,PCBP2的mRNA和蛋白表达水平显著下调,DBV感染诱导典型铁死亡特征(线粒体嵴减少、肿胀)。通过qRT-PCR和Western blot验证,PCBP2敲低和过表达的THP-1细胞系构建成功,PCBP2敲低下调了铁死亡相关基因SLC7A11和GPX4的表达,导致ROS和Fe^(2+)水平升高;相反,PCBP2过表达使得SLC7A11和GPX4的表达水平升高,ROS和Fe^(2+)的水平降低。半数组织培养感染剂量与蛋白水平的检测进一步证实:铁死亡诱导剂可部分抵消PCBP2过表达促病毒复制的效应,铁死亡抑制剂可部分逆转PCBP2敲低抑制病毒复制的效应。结论:研究发现PCBP2可以通过维持SLC7A11/GPX4系统功能抑制铁死亡,从而限制DBV复制。这不仅阐明了PCBP2在DBV感染中的调控作用,为发热伴血小板减少综合征(severe fever with thrombocytope-nia syndrome,SFTS)的发病机制提供了新见解,同时靶向PCBP2-铁死亡通路可能成为SFTS治疗的潜在策略,为抗病毒药物的研发提供新思路。展开更多
基金supported by funds from the Conselho Nacional de Desenvolvimento Cientifico e Tecnologico do Brasil(CNPq)(312286/2023-6,307201/2023-6,and Instituto Nacional Saude Cerebral INSC,No.406020/2022-1)Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior(CAPES)Fundacao de Amparo a Pesquisa do Estado do Rio de Janeiro-FAPERJ(E-26/010.002260/2019,E-26/010.001652/2019,E-26/010.101036/2018,E-26/202.774/2018,E-26/210.240/2020,E-26/211.138/2021,26/210.823/2021,E-26/211.325/2021,E-26/210.779/2021,E-26/201.086/2022,E-26/210.312/2022,E-26/203.262/2023,E-26/200.195/2023)(to LEBS)。
文摘Recent increases in infectious diseases affecting the central nervous system have raised concerns about their role in neuroinflammation and neurodegeneration.Viral pathogens or their products can invade the central nervous system and cause damage,leading to meningitis,encephalitis,meningoencephalitis,myelitis,or post-infectious demyelinating diseases.Although neuroinflammation initially has a protective function,chronic inflammation can contribute to the development of neurodegenerative diseases.Mechanisms such as protein aggregation and cellular disturbances are implicated with specific viruses such as herpes simplex virus type 1 and Epstein-Barr virus being associated with Alzheimer's disease and multiple sclerosis,respectively.Extracellular nucleotides,particularly adenosine triphosphate and its metabolites are released from activated,infected,and dying cells,acting as alarmins mediating neuroinflammation and neurodegeneration.When viruses infect central nervous system cells,adenosine triphosphate is released as an alarmin,triggering inflammatory responses.This process is mediated by purinergic receptors,divided into two families:P1,which responds to adenosine,and P2,activated by adenosine triphosphate and other nucleotides.This review highlights how specific viruses,such as human immunodeficiency virus type 1,Theiler's murine encephalomyelitis virus,herpes simplex virus type 1,Epstein-Barr virus,dengue virus,Zika virus,and severe acute respiratory syndrome coronavirus 2,can initiate inflammatory responses through the release of extracellular nucleotides,particularly adenosine triphosphate,which act as critical mediators in the progression of neuroinflammation and neurodegenerative disorders.A better understanding of purinergic signaling pathways in these diseases may suggest new potential therapeutic strategies for targeting neuroinflammation to mitigate the long-term consequences of viral infections in the central nervous system.
文摘目的:探究多聚胞嘧啶结合蛋白2[poly(C)-binding protein 2,PCBP2]如何通过调节铁死亡参与大别班达病毒(Dabie Banda virus,DBV)感染后的致病过程及其作用机制。方法:以人单核细胞系THP-1为模型,采用qRT-PCR和Western blot技术检测DBV感染的THP-1细胞中PCBP2的mRNA及蛋白表达水平。通过透射电镜观察病毒感染下的线粒体结构变化,在THP-1细胞中构建了慢病毒介导的PCBP2过表达和敲低稳转细胞系。FerroOrange荧光探针检测Fe^(2+)水平,2,7-二氯荧光素二乙酸酯(2,7-dichlorofluorescein diacetate,DCFH-DA)探针测定活性氧(reactive oxygen species,ROS)水平,Western blot检测铁死亡相关溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)和谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)蛋白表达,以评估PCBP2调控对铁死亡的影响。使用铁死亡诱导剂(RSL3、erastin)和抑制剂(Fer-1、Lip-1)处理细胞,qRT-PCR和免疫荧光检测病毒复制水平变化,探索PCBP2是否可以通过调控铁死亡影响DBV复制。结果:在DBV感染的细胞模型中,PCBP2的mRNA和蛋白表达水平显著下调,DBV感染诱导典型铁死亡特征(线粒体嵴减少、肿胀)。通过qRT-PCR和Western blot验证,PCBP2敲低和过表达的THP-1细胞系构建成功,PCBP2敲低下调了铁死亡相关基因SLC7A11和GPX4的表达,导致ROS和Fe^(2+)水平升高;相反,PCBP2过表达使得SLC7A11和GPX4的表达水平升高,ROS和Fe^(2+)的水平降低。半数组织培养感染剂量与蛋白水平的检测进一步证实:铁死亡诱导剂可部分抵消PCBP2过表达促病毒复制的效应,铁死亡抑制剂可部分逆转PCBP2敲低抑制病毒复制的效应。结论:研究发现PCBP2可以通过维持SLC7A11/GPX4系统功能抑制铁死亡,从而限制DBV复制。这不仅阐明了PCBP2在DBV感染中的调控作用,为发热伴血小板减少综合征(severe fever with thrombocytope-nia syndrome,SFTS)的发病机制提供了新见解,同时靶向PCBP2-铁死亡通路可能成为SFTS治疗的潜在策略,为抗病毒药物的研发提供新思路。
