Neuroserpin, a secreted protein that belongs to the serpin superfamily of serine protease inhibitors, is highly expressed in the central nervous system and plays multiple roles in brain development and pathology. As a...Neuroserpin, a secreted protein that belongs to the serpin superfamily of serine protease inhibitors, is highly expressed in the central nervous system and plays multiple roles in brain development and pathology. As a natural inhibitor of recombinant tissue plasminogen activator, neuroserpin inhibits the increased activity of tissue plasminogen activator in ischemic conditions and extends the therapeutic windows of tissue plasminogen activator for brain ischemia. However, the neuroprotective mechanism of neuroserpin against ischemic stroke remains unclear. In this study, we used a mouse model of middle cerebral artery occlusion and oxygen-glucose deprivation/reperfusion-injured cortical neurons as in vivo and in vitro ischemia-reperfusion models, respectively. The models were used to investigate the neuroprotective effects of neuroserpin. Our findings revealed that endoplasmic reticulum stress was promptly triggered following ischemia, initially manifesting as the acute activation of endoplasmic reticulum stress transmembrane sensors and the suppression of protein synthesis, which was followed by a later apoptotic response. Notably, ischemic stroke markedly downregulated the expression of neuroserpin in cortical neurons. Exogenous neuroserpin reversed the activation of multiple endoplasmic reticulum stress signaling molecules, the reduction in protein synthesis, and the upregulation of apoptotic transcription factors. This led to a reduction in neuronal death induced by oxygen/glucose deprivation and reperfusion, as well as decreased cerebral infarction and neurological dysfunction in mice with middle cerebral artery occlusion. However, the neuroprotective effects of neuroserpin were markedly inhibited by endoplasmic reticulum stress activators thapsigargin and tunicamycin. Our findings demonstrate that neuroserpin exerts neuroprotective effects on ischemic stroke by suppressing endoplasmic reticulum stress.展开更多
A number of studies have confirmed the existence of tissue-type plasminogen activator-independent roles of neuroserpin, a member of the serine protease inhibitor superfamily. In this review article, we aim to clarify ...A number of studies have confirmed the existence of tissue-type plasminogen activator-independent roles of neuroserpin, a member of the serine protease inhibitor superfamily. In this review article, we aim to clarify this role. These unique roles of neuroserpin are involved in its neuroprotective effect during ischemic brain injury, its regulation of tumorigenesis, and the mediation of emotion and cognition through the inhibition of urokinase-type plasminogen activator and fibrinolysin, modification of Th cells, reducing plaque formation, promoting process growth and intracellular adhesion, and alterina the expression of cadherin and nuclear factor kaooa B.展开更多
Tissue plasminogen activator is usually used for the treatment of acute ischemic stroke,but the role of endogenous tissue plasminogen activator in traumatic brain injury has been rarely reported.A rat model of traumat...Tissue plasminogen activator is usually used for the treatment of acute ischemic stroke,but the role of endogenous tissue plasminogen activator in traumatic brain injury has been rarely reported.A rat model of traumatic brain injury was established by weight-drop method.The tissue plasminogen activator inhibitor neuroserpin(5μL,0.25 mg/mL)was injected into the lateral ventricle.Neurological function was assessed by neurological severity score.Neuronal and axonal injuries were assessed by hematoxylin-eosin staining and Bielschowsky silver staining.Protein level of endogenous tissue plasminogen activator was analyzed by western blot assay.Apoptotic marker cleaved caspase-3,neuronal marker neurofilament light chain,astrocyte marker glial fibrillary acidic protein and microglial marker Iba-1 were analyzed by immunohistochemical staining.Apoptotic cell types were detected by immunofluorescence double labeling.Apoptotic cells in the damaged cortex were detected by terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP-biotin nick-end labeling staining.Degenerating neurons in the damaged cortex were detected by Fluoro-Jade B staining.Expression of tissue plasminogen activator was increased at 6 hours,and peaked at 3 days after traumatic brain injury.Neuronal apoptosis and axonal injury were detected after traumatic brain injury.Moreover,neuroserpin enhanced neuronal apoptosis,neuronal injury and axonal injury,and activated microglia and astrocytes.Neuroserpin further deteriorated neurobehavioral function in rats with traumatic brain injury.Our findings confirm that inhibition of endogenous tissue plasminogen activator aggravates neuronal apoptosis and axonal injury after traumatic brain injury,and activates microglia and astrocytes.This study was approved by the Biomedical Ethics Committee of Animal Experiments of Shaanxi Province of China in June 2015.展开更多
Dementia is a clinical syndrome that affects approximately 47 million people worldwide and is characterized by progressive and irreversible decline of cognitive,behavioral and sesorimotor functions.Alzheimer’s diseas...Dementia is a clinical syndrome that affects approximately 47 million people worldwide and is characterized by progressive and irreversible decline of cognitive,behavioral and sesorimotor functions.Alzheimer’s disease(AD)accounts for approximately 60–80%of all cases of dementia,and neuropathologically is characterized by extracellular deposits of insoluble amyloid-β(Aβ)and intracellular aggregates of hyperphosphorylated tau.Significantly,although for a long time it was believed that the extracellular accumulation of Aβwas the culprit of the symptoms observed in these patients,more recent studies have shown that cognitive decline in people suffering this disease is associated with soluble Aβ-induced synaptic dysfunction instead of the formation of insoluble Aβ-containing extracellular plaques.These observations are translationally relevant because soluble Aβ-induced synaptic dysfunction is an early event in AD that precedes neuronal death,and thus is amenable to therapeutic interventions to prevent cognitive decline before the progression to irreversible brain damage.The plasminogen activating(PA)system is an enzymatic cascade that triggers the degradation of fibrin by catalyzing the conversion of plasminogen into plasmin via two serine proteinases:tissue-type plasminogen activator(tPA)and urokinase-type plasminogen activator(uPA).Experimental evidence reported over the last three decades has shown that tPA and uPA play a role in the pathogenesis of AD.However,these studies have focused on the ability of these plasminogen activators to trigger plasmin-induced cleavage of insoluble Aβ-containing extracellular plaques.In contrast,recent evidence indicates that activity-dependent release of uPA from the presynaptic terminal of cerebral cortical neurons protects the synapse from the deleterious effects of soluble Aβvia a mechanism that does not require plasmin generation or the cleavage of Aβfibrils.Below we discuss the role of the PA system in the pathogenesis of AD and the translational relevance of data published to this date.展开更多
Protein quality control involves many processes that jointly act to regulate the expression, localization, turnover, and degradation of proteins, and has been highlighted in recent studies as critical to the different...Protein quality control involves many processes that jointly act to regulate the expression, localization, turnover, and degradation of proteins, and has been highlighted in recent studies as critical to the differentiation of stem cells during regeneration. The roles of constitutively secreted extracellular chaperones in neuronal injury and disease are poorly understood. Extracellular chaperones are multifunctional proteins expressed by many cell types, including those of the nervous system, known to facilitate protein quality control processes. These molecules exert pleiotropic effects and have been implicated as playing important protective roles in a variety of stress conditions, including tissue damage, infections, and local tissue inflammation. This article aims to provide a critical review of what is currently known about the functions of extracellular chaperones in neuronal repair and regeneration and highlight future directions for this important research area. We review what is known of four constitutively secreted extracellular chaperones directly implicated in processes of neuronal damage and repair, including transthyretin, clusterin, α2-macroglobulin, and neuroserpin, and propose that investigation into the effects of these and other extracellular chaperones on neuronal repair and regeneration has the potential to yield valuable new therapies.展开更多
目的探讨神经源性丝氨酸蛋白酶抑制剂(neuroserpin,NSP)对急性缺血性脑卒中时使用重组人组织型纤溶酶原激活物(tissue plasminogen activator,tPA)治疗的影响和机制。方法构造大鼠大脑中动脉阻塞(middle cerebral artery occlusion,MCAO...目的探讨神经源性丝氨酸蛋白酶抑制剂(neuroserpin,NSP)对急性缺血性脑卒中时使用重组人组织型纤溶酶原激活物(tissue plasminogen activator,tPA)治疗的影响和机制。方法构造大鼠大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)模型。将大鼠随机分为假手术组、MCAO组、1 h tPA组(再灌注1 h MCAO+tPA)、1h tPA+NSP组(再灌注1 h MCAO+tPA+NSP)、4 h tPA组(再灌注4 h MCAO+tPA)及4 h tPA+NSP组(再灌注4 h MCAO+tPA+NSP)。采用改良神经严重性评分(modified neurologic severity score,mNSS)、Rota-Rod电机测试评估大鼠神经功能改变,采用干湿法和TTC染色评估大鼠脑水肿和脑梗死面积,采用伊文思蓝染色法、苏木精-伊红(Hematoxylin Eosin,HE)染色法检测大鼠血脑屏障完整性,采用酶联免疫吸附试验法检测脑组织丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)的表达,采用蛋白质印迹法和甲苯胺蓝染色检测脑组织中半胱门冬氨酸蛋白酶(cysteinyl aspartate specific proteinase,Caspase)-3、B细胞淋巴瘤因子-2(B-cell lymphoma factor 2,Bcl-2)的表达以及脑组织细胞凋亡情况。结果mNSS结果显示,1h tPA+NSP组及4 h tPA+NSP组(联合干预组)分别低于1 h tPA组和4 h tPA组(P<0.05)。Rota-Rod电机测试结果显示,各联合干预组分别长于1 h tPA组和4 htPA组(P<0.05)。各联合干预组脑水肿体积和脑梗死面积均优于相同再灌注时间单纯tPA干预组。各联合干预组MDA含量低于相同再灌注时间单纯tPA干预组(P<0.05),各联合干预组SOD及GSH-Px活性高于相同再灌注时间单纯tPA干预组(P<0.05)。各联合干预组Caspase-3表达低于相同再灌注时间单纯tPA干预组(P<0.05)。各联合干预组Bcl-2表达高于相同再灌注时间单纯tPA干预组(P<0.05)。结论NSP可以改善tPA干预后引起的神经功能缺损,其机制可能是NSP减弱了脑缺血时血脑屏障的损伤和氧化应激反应。展开更多
神经丝酶抑制蛋白(neuroserpin,NSP)是组织型纤维蛋白溶酶原激活剂(tissue-type plasminogen activator,tPA)在神经系统中的特异性抑制剂,主要参与并调节学习、记忆及行为等活动,在神经发育及重塑中发挥着重要作用。N S P在缺血性脑损...神经丝酶抑制蛋白(neuroserpin,NSP)是组织型纤维蛋白溶酶原激活剂(tissue-type plasminogen activator,tPA)在神经系统中的特异性抑制剂,主要参与并调节学习、记忆及行为等活动,在神经发育及重塑中发挥着重要作用。N S P在缺血性脑损伤及癫痫发生时具有神经保护作用。N S P突变体可以导致一种常染色体显性遗传性痴呆,即N S P包含体家族性脑病(familial encephalopathy with neuroserpin inclusion bodies,FENIB)。本文就该病发生的分子结构基础,突变型和表现型之间的对应关系,以及治疗进展三方面进行综述。展开更多
基金supported in part by the National Key Research&Development Program of China,No. 2022YFA1104900 (to LS)the National Natural Science Foundation of China,Nos. 82371175, 82071535 (both to LS), 82101614 (to YP)+5 种基金the International Science and Technology Cooperation Projects of Guangdong Province,No. 2023A0505050121 (to LS)Guangdong Basic and Applied Basic Research Foundation,Nos. 2022B1515130007 (to LS), 2023A1515030012 (to SZ), 2022A1515010666 (to WL)the Science and Technology Program of Guangzhou,Nos. 202102070001 (to LS), 202201010041 (to YP)Shenzhen Basic Research Grant,Nos. JCYJ20200109140414636, JCYJ20230807145103007 (both to WL)awarded a Royal Society Newton Advanced Fellowship,No. AOMS-NAF0051003in collaboration with Zoltán Molnár,Department of Physiology,Anatomy and Genetics,University of Oxford (2017–2021)。
文摘Neuroserpin, a secreted protein that belongs to the serpin superfamily of serine protease inhibitors, is highly expressed in the central nervous system and plays multiple roles in brain development and pathology. As a natural inhibitor of recombinant tissue plasminogen activator, neuroserpin inhibits the increased activity of tissue plasminogen activator in ischemic conditions and extends the therapeutic windows of tissue plasminogen activator for brain ischemia. However, the neuroprotective mechanism of neuroserpin against ischemic stroke remains unclear. In this study, we used a mouse model of middle cerebral artery occlusion and oxygen-glucose deprivation/reperfusion-injured cortical neurons as in vivo and in vitro ischemia-reperfusion models, respectively. The models were used to investigate the neuroprotective effects of neuroserpin. Our findings revealed that endoplasmic reticulum stress was promptly triggered following ischemia, initially manifesting as the acute activation of endoplasmic reticulum stress transmembrane sensors and the suppression of protein synthesis, which was followed by a later apoptotic response. Notably, ischemic stroke markedly downregulated the expression of neuroserpin in cortical neurons. Exogenous neuroserpin reversed the activation of multiple endoplasmic reticulum stress signaling molecules, the reduction in protein synthesis, and the upregulation of apoptotic transcription factors. This led to a reduction in neuronal death induced by oxygen/glucose deprivation and reperfusion, as well as decreased cerebral infarction and neurological dysfunction in mice with middle cerebral artery occlusion. However, the neuroprotective effects of neuroserpin were markedly inhibited by endoplasmic reticulum stress activators thapsigargin and tunicamycin. Our findings demonstrate that neuroserpin exerts neuroprotective effects on ischemic stroke by suppressing endoplasmic reticulum stress.
基金supported by the National Natural Science Foundation of China, No. 30700908, 30772343 and 30973215
文摘A number of studies have confirmed the existence of tissue-type plasminogen activator-independent roles of neuroserpin, a member of the serine protease inhibitor superfamily. In this review article, we aim to clarify this role. These unique roles of neuroserpin are involved in its neuroprotective effect during ischemic brain injury, its regulation of tumorigenesis, and the mediation of emotion and cognition through the inhibition of urokinase-type plasminogen activator and fibrinolysin, modification of Th cells, reducing plaque formation, promoting process growth and intracellular adhesion, and alterina the expression of cadherin and nuclear factor kaooa B.
文摘Tissue plasminogen activator is usually used for the treatment of acute ischemic stroke,but the role of endogenous tissue plasminogen activator in traumatic brain injury has been rarely reported.A rat model of traumatic brain injury was established by weight-drop method.The tissue plasminogen activator inhibitor neuroserpin(5μL,0.25 mg/mL)was injected into the lateral ventricle.Neurological function was assessed by neurological severity score.Neuronal and axonal injuries were assessed by hematoxylin-eosin staining and Bielschowsky silver staining.Protein level of endogenous tissue plasminogen activator was analyzed by western blot assay.Apoptotic marker cleaved caspase-3,neuronal marker neurofilament light chain,astrocyte marker glial fibrillary acidic protein and microglial marker Iba-1 were analyzed by immunohistochemical staining.Apoptotic cell types were detected by immunofluorescence double labeling.Apoptotic cells in the damaged cortex were detected by terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP-biotin nick-end labeling staining.Degenerating neurons in the damaged cortex were detected by Fluoro-Jade B staining.Expression of tissue plasminogen activator was increased at 6 hours,and peaked at 3 days after traumatic brain injury.Neuronal apoptosis and axonal injury were detected after traumatic brain injury.Moreover,neuroserpin enhanced neuronal apoptosis,neuronal injury and axonal injury,and activated microglia and astrocytes.Neuroserpin further deteriorated neurobehavioral function in rats with traumatic brain injury.Our findings confirm that inhibition of endogenous tissue plasminogen activator aggravates neuronal apoptosis and axonal injury after traumatic brain injury,and activates microglia and astrocytes.This study was approved by the Biomedical Ethics Committee of Animal Experiments of Shaanxi Province of China in June 2015.
基金This work was supported in part by National Institutes of Health Grant NS-NS091201(to MY)and VA MERIT Award IO1BX003441(to MY).
文摘Dementia is a clinical syndrome that affects approximately 47 million people worldwide and is characterized by progressive and irreversible decline of cognitive,behavioral and sesorimotor functions.Alzheimer’s disease(AD)accounts for approximately 60–80%of all cases of dementia,and neuropathologically is characterized by extracellular deposits of insoluble amyloid-β(Aβ)and intracellular aggregates of hyperphosphorylated tau.Significantly,although for a long time it was believed that the extracellular accumulation of Aβwas the culprit of the symptoms observed in these patients,more recent studies have shown that cognitive decline in people suffering this disease is associated with soluble Aβ-induced synaptic dysfunction instead of the formation of insoluble Aβ-containing extracellular plaques.These observations are translationally relevant because soluble Aβ-induced synaptic dysfunction is an early event in AD that precedes neuronal death,and thus is amenable to therapeutic interventions to prevent cognitive decline before the progression to irreversible brain damage.The plasminogen activating(PA)system is an enzymatic cascade that triggers the degradation of fibrin by catalyzing the conversion of plasminogen into plasmin via two serine proteinases:tissue-type plasminogen activator(tPA)and urokinase-type plasminogen activator(uPA).Experimental evidence reported over the last three decades has shown that tPA and uPA play a role in the pathogenesis of AD.However,these studies have focused on the ability of these plasminogen activators to trigger plasmin-induced cleavage of insoluble Aβ-containing extracellular plaques.In contrast,recent evidence indicates that activity-dependent release of uPA from the presynaptic terminal of cerebral cortical neurons protects the synapse from the deleterious effects of soluble Aβvia a mechanism that does not require plasmin generation or the cleavage of Aβfibrils.Below we discuss the role of the PA system in the pathogenesis of AD and the translational relevance of data published to this date.
文摘Protein quality control involves many processes that jointly act to regulate the expression, localization, turnover, and degradation of proteins, and has been highlighted in recent studies as critical to the differentiation of stem cells during regeneration. The roles of constitutively secreted extracellular chaperones in neuronal injury and disease are poorly understood. Extracellular chaperones are multifunctional proteins expressed by many cell types, including those of the nervous system, known to facilitate protein quality control processes. These molecules exert pleiotropic effects and have been implicated as playing important protective roles in a variety of stress conditions, including tissue damage, infections, and local tissue inflammation. This article aims to provide a critical review of what is currently known about the functions of extracellular chaperones in neuronal repair and regeneration and highlight future directions for this important research area. We review what is known of four constitutively secreted extracellular chaperones directly implicated in processes of neuronal damage and repair, including transthyretin, clusterin, α2-macroglobulin, and neuroserpin, and propose that investigation into the effects of these and other extracellular chaperones on neuronal repair and regeneration has the potential to yield valuable new therapies.
文摘目的探讨神经源性丝氨酸蛋白酶抑制剂(neuroserpin,NSP)对急性缺血性脑卒中时使用重组人组织型纤溶酶原激活物(tissue plasminogen activator,tPA)治疗的影响和机制。方法构造大鼠大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)模型。将大鼠随机分为假手术组、MCAO组、1 h tPA组(再灌注1 h MCAO+tPA)、1h tPA+NSP组(再灌注1 h MCAO+tPA+NSP)、4 h tPA组(再灌注4 h MCAO+tPA)及4 h tPA+NSP组(再灌注4 h MCAO+tPA+NSP)。采用改良神经严重性评分(modified neurologic severity score,mNSS)、Rota-Rod电机测试评估大鼠神经功能改变,采用干湿法和TTC染色评估大鼠脑水肿和脑梗死面积,采用伊文思蓝染色法、苏木精-伊红(Hematoxylin Eosin,HE)染色法检测大鼠血脑屏障完整性,采用酶联免疫吸附试验法检测脑组织丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)的表达,采用蛋白质印迹法和甲苯胺蓝染色检测脑组织中半胱门冬氨酸蛋白酶(cysteinyl aspartate specific proteinase,Caspase)-3、B细胞淋巴瘤因子-2(B-cell lymphoma factor 2,Bcl-2)的表达以及脑组织细胞凋亡情况。结果mNSS结果显示,1h tPA+NSP组及4 h tPA+NSP组(联合干预组)分别低于1 h tPA组和4 h tPA组(P<0.05)。Rota-Rod电机测试结果显示,各联合干预组分别长于1 h tPA组和4 htPA组(P<0.05)。各联合干预组脑水肿体积和脑梗死面积均优于相同再灌注时间单纯tPA干预组。各联合干预组MDA含量低于相同再灌注时间单纯tPA干预组(P<0.05),各联合干预组SOD及GSH-Px活性高于相同再灌注时间单纯tPA干预组(P<0.05)。各联合干预组Caspase-3表达低于相同再灌注时间单纯tPA干预组(P<0.05)。各联合干预组Bcl-2表达高于相同再灌注时间单纯tPA干预组(P<0.05)。结论NSP可以改善tPA干预后引起的神经功能缺损,其机制可能是NSP减弱了脑缺血时血脑屏障的损伤和氧化应激反应。
文摘神经丝酶抑制蛋白(neuroserpin,NSP)是组织型纤维蛋白溶酶原激活剂(tissue-type plasminogen activator,tPA)在神经系统中的特异性抑制剂,主要参与并调节学习、记忆及行为等活动,在神经发育及重塑中发挥着重要作用。N S P在缺血性脑损伤及癫痫发生时具有神经保护作用。N S P突变体可以导致一种常染色体显性遗传性痴呆,即N S P包含体家族性脑病(familial encephalopathy with neuroserpin inclusion bodies,FENIB)。本文就该病发生的分子结构基础,突变型和表现型之间的对应关系,以及治疗进展三方面进行综述。
