Parkinson's disease is primarily caused by the loss of dopaminergic neurons in the substantia nigra compacta.Ferroptosis,a novel form of regulated cell death characterized by iron accumulation and lipid peroxidati...Parkinson's disease is primarily caused by the loss of dopaminergic neurons in the substantia nigra compacta.Ferroptosis,a novel form of regulated cell death characterized by iron accumulation and lipid peroxidation,plays a vital role in the death of dopaminergic neurons.However,the molecular mechanisms underlying ferroptosis in dopaminergic neurons have not yet been completely elucidated.NADPH oxidase 4 is related to oxidative stress,however,whether it regulates dopaminergic neuronal ferroptosis remains unknown.The aim of this study was to determine whether NADPH oxidase 4 is involved in dopaminergic neuronal ferroptosis,and if so,by what mechanism.We found that the transcriptional regulator activating transcription factor 3 increased NADPH oxidase 4 expression in dopaminergic neurons and astrocytes in an 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine-induced Parkinson's disease model.NADPH oxidase 4 inhibition improved the behavioral impairments observed in the Parkinson's disease model animals and reduced the death of dopaminergic neurons.Moreover,NADPH oxidase 4 inhibition reduced lipid peroxidation and iron accumulation in the substantia nigra of the Parkinson's disease model animals.Mechanistically,we found that NADPH oxidase 4 interacted with activated protein kinase Cαto prevent ferroptosis of dopaminergic neurons.Furthermore,by lowering the astrocytic lipocalin-2 expression,NADPH oxidase 4 inhibition reduced 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine-induced neuroinflammation.These findings demonstrate that NADPH oxidase 4 promotes ferroptosis of dopaminergic neurons and neuroinflammation,which contribute to dopaminergic neuron death,suggesting that NADPH oxidase 4 is a possible therapeutic target for Parkinson's disease.展开更多
目的:研究NADPH氧化酶4(Nox4)在骨癌痛(CIBP)大鼠背根神经节(DRG)中的表达。方法:48只成年雄性SD大鼠随机分为4组:假手术组(Sham)、骨癌痛组(CIBP)、空病毒对照组(Lenti-NC)与Nox4基因沉默组(Lenti-shNox4)。通过鞘内注射方法给予大鼠...目的:研究NADPH氧化酶4(Nox4)在骨癌痛(CIBP)大鼠背根神经节(DRG)中的表达。方法:48只成年雄性SD大鼠随机分为4组:假手术组(Sham)、骨癌痛组(CIBP)、空病毒对照组(Lenti-NC)与Nox4基因沉默组(Lenti-shNox4)。通过鞘内注射方法给予大鼠重组慢病毒处理,通过胫骨骨髓腔接种Walker256细胞的方法建立大鼠骨癌痛模型,通过胫骨HE染色及行为学检测鉴定大鼠CIBP模型,应用real time RT-PCR检测大鼠DRG中Nox4 mRNA的表达,利用Western Blot和免疫组织荧光染色检测Nox4蛋白在大鼠DRG中的表达和细胞定位,利用商品化试剂盒检测大鼠DRG中H2O2的含量,应用real time RT-PCR和Western Blot检测Nox4基因沉默对大鼠DRG中神经型一氧化氮合酶(n NOS)、超氧化物歧化酶(SOD)及N-甲基-D-天冬氨酸受体2D(NMDAR2D)和γ-氨基丁酸A型γ2受体(GABAA-γ2)表达的影响。结果:大鼠接种Walker256细胞可造成胫骨破坏及肿瘤细胞生长,PWT减少和TWL缩短;在骨癌痛大鼠DRG中,Nox4的表达显著升高;免疫组织荧光染色结果表明Nox4主要表达于DRG小胶质细胞中;敲减DRG中Nox4表达能减轻骨癌痛大鼠的机械痛敏和热痛敏,降低DRG中H2O2的水平,下调nNOS和NMDAR2D表达,上调SOD和GABAA-γ2表达。结论:敲减DRG中Nox4表达可通过减轻氧化应激反应缓解大鼠骨癌痛。展开更多
基金supported by the National Natural Science Foundation of China,Nos.82271444(to JP),82271268(to BZ),and 82001346(to YL)the National Key Research and Development Program of China,No.2022YFE0210100(to BZ)。
文摘Parkinson's disease is primarily caused by the loss of dopaminergic neurons in the substantia nigra compacta.Ferroptosis,a novel form of regulated cell death characterized by iron accumulation and lipid peroxidation,plays a vital role in the death of dopaminergic neurons.However,the molecular mechanisms underlying ferroptosis in dopaminergic neurons have not yet been completely elucidated.NADPH oxidase 4 is related to oxidative stress,however,whether it regulates dopaminergic neuronal ferroptosis remains unknown.The aim of this study was to determine whether NADPH oxidase 4 is involved in dopaminergic neuronal ferroptosis,and if so,by what mechanism.We found that the transcriptional regulator activating transcription factor 3 increased NADPH oxidase 4 expression in dopaminergic neurons and astrocytes in an 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine-induced Parkinson's disease model.NADPH oxidase 4 inhibition improved the behavioral impairments observed in the Parkinson's disease model animals and reduced the death of dopaminergic neurons.Moreover,NADPH oxidase 4 inhibition reduced lipid peroxidation and iron accumulation in the substantia nigra of the Parkinson's disease model animals.Mechanistically,we found that NADPH oxidase 4 interacted with activated protein kinase Cαto prevent ferroptosis of dopaminergic neurons.Furthermore,by lowering the astrocytic lipocalin-2 expression,NADPH oxidase 4 inhibition reduced 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine-induced neuroinflammation.These findings demonstrate that NADPH oxidase 4 promotes ferroptosis of dopaminergic neurons and neuroinflammation,which contribute to dopaminergic neuron death,suggesting that NADPH oxidase 4 is a possible therapeutic target for Parkinson's disease.
文摘目的:研究NADPH氧化酶4(Nox4)在骨癌痛(CIBP)大鼠背根神经节(DRG)中的表达。方法:48只成年雄性SD大鼠随机分为4组:假手术组(Sham)、骨癌痛组(CIBP)、空病毒对照组(Lenti-NC)与Nox4基因沉默组(Lenti-shNox4)。通过鞘内注射方法给予大鼠重组慢病毒处理,通过胫骨骨髓腔接种Walker256细胞的方法建立大鼠骨癌痛模型,通过胫骨HE染色及行为学检测鉴定大鼠CIBP模型,应用real time RT-PCR检测大鼠DRG中Nox4 mRNA的表达,利用Western Blot和免疫组织荧光染色检测Nox4蛋白在大鼠DRG中的表达和细胞定位,利用商品化试剂盒检测大鼠DRG中H2O2的含量,应用real time RT-PCR和Western Blot检测Nox4基因沉默对大鼠DRG中神经型一氧化氮合酶(n NOS)、超氧化物歧化酶(SOD)及N-甲基-D-天冬氨酸受体2D(NMDAR2D)和γ-氨基丁酸A型γ2受体(GABAA-γ2)表达的影响。结果:大鼠接种Walker256细胞可造成胫骨破坏及肿瘤细胞生长,PWT减少和TWL缩短;在骨癌痛大鼠DRG中,Nox4的表达显著升高;免疫组织荧光染色结果表明Nox4主要表达于DRG小胶质细胞中;敲减DRG中Nox4表达能减轻骨癌痛大鼠的机械痛敏和热痛敏,降低DRG中H2O2的水平,下调nNOS和NMDAR2D表达,上调SOD和GABAA-γ2表达。结论:敲减DRG中Nox4表达可通过减轻氧化应激反应缓解大鼠骨癌痛。
