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1.
Neuron ; 110(18): 2929-2948.e8, 2022 09 21.
Artículo en Inglés | MEDLINE | ID: mdl-35882228

RESUMEN

Tau aggregation in neurofibrillary tangles (NFTs) is closely associated with neurodegeneration and cognitive decline in Alzheimer's disease (AD). However, the molecular signatures that distinguish between aggregation-prone and aggregation-resistant cell states are unknown. We developed methods for the high-throughput isolation and transcriptome profiling of single somas with NFTs from the human AD brain, quantified the susceptibility of 20 neocortical subtypes for NFT formation and death, and identified both shared and cell-type-specific signatures. NFT-bearing neurons shared a marked upregulation of synaptic transmission-related genes, including a core set of 63 genes enriched for synaptic vesicle cycling. Oxidative phosphorylation and mitochondrial dysfunction were highly cell-type dependent. Apoptosis was only modestly enriched, and the susceptibilities of NFT-bearing and NFT-free neurons for death were highly similar. Our analysis suggests that NFTs represent cell-type-specific responses to stress and synaptic dysfunction. We provide a resource for biomarker discovery and the investigation of tau-dependent and tau-independent mechanisms of neurodegeneration.


Asunto(s)
Enfermedad de Alzheimer , Ovillos Neurofibrilares , Enfermedad de Alzheimer/genética , Enfermedad de Alzheimer/metabolismo , Biomarcadores/metabolismo , Encéfalo/metabolismo , Humanos , Ovillos Neurofibrilares/metabolismo , Proteínas tau/genética , Proteínas tau/metabolismo
2.
Mol Neurodegener ; 10: 24, 2015 Jun 24.
Artículo en Inglés | MEDLINE | ID: mdl-26104027

RESUMEN

BACKGROUND: Cerebral infarction due to thrombosis leads to the most common type of stroke and a likely cause of age-related cognitive decline and dementia. Endothelial nitric oxide synthase (eNOS) generates NO, which plays a crucial role in maintaining vascular function and exerting an antithrombotic action. Reduced eNOS expression and eNOS polymorphisms have been associated with stroke and Alzheimer's disease (AD), the most common type of dementia associated with neurovascular dysfunction. However, direct proof of such association is lacking. Since there are no reports of complete eNOS deficiency in humans, we used heterozygous eNOS(+/-) mice to mimic partial deficiency of eNOS, and determine its impact on cerebrovascular pathology and perfusion of cerebral vessels. RESULTS: Combining cerebral angiography with immunohistochemistry, we found thrombotic cerebral infarctions in eNOS(+/-) mice as early as 3-6 months of age but not in eNOS(+/+) mice at any age. Remarkably, vascular occlusions in eNOS(+/-) mice were found almost exclusively in three areas: temporoparietal and retrosplenial granular cortexes, and hippocampus this distribution precisely matching the hypoperfused areas identified in preclinical AD patients. Moreover, progressive cerebral amyloid angiopaphy (CAA), blood brain barrier (BBB) breakdown, and cognitive impairment were also detected in aged eNOS(+/-) mice. CONCLUSIONS: These data provide for the first time the evidence that partial eNOS deficiency results in spontaneous thrombotic cerebral infarctions that increase with age, leading to progressive CAA and cognitive impairments. We thus conclude that eNOS(+/-) mouse may represent an ideal model of ischemic stroke to address early and progressive damage in spontaneously-evolving chronic cerebral ischemia and thus, study vascular mechanisms contributing to vascular dementia and AD.


Asunto(s)
Barrera Hematoencefálica/fisiología , Angiopatía Amiloide Cerebral/genética , Infarto Cerebral/genética , Trastornos del Conocimiento/genética , Óxido Nítrico Sintasa de Tipo III/fisiología , Envejecimiento , Animales , Angiopatía Amiloide Cerebral/patología , Infarto Cerebral/enzimología , Infarto Cerebral/patología , Infarto Cerebral/psicología , Circulación Cerebrovascular , Trastornos del Conocimiento/enzimología , Trastornos del Conocimiento/patología , Progresión de la Enfermedad , Endotelio Vascular/fisiopatología , Regulación de la Expresión Génica , Heterocigoto , Hipocampo/irrigación sanguínea , Hipocampo/enzimología , Hipocampo/patología , Aprendizaje por Laberinto , Trastornos de la Memoria/enzimología , Trastornos de la Memoria/genética , Trastornos de la Memoria/patología , Ratones , Ratones Mutantes Neurológicos , Proteínas del Tejido Nervioso/biosíntesis , Proteínas del Tejido Nervioso/genética , Óxido Nítrico Sintasa de Tipo III/deficiencia , Óxido Nítrico Sintasa de Tipo III/genética , Especificidad de Órganos , Lóbulo Parietal/irrigación sanguínea , Lóbulo Parietal/enzimología , Lóbulo Parietal/patología , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Lóbulo Temporal/irrigación sanguínea , Lóbulo Temporal/enzimología , Lóbulo Temporal/patología
3.
PLoS One ; 8(5): e64389, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23717608

RESUMEN

Gene regulation remains one of the major challenges for gene therapy in clinical trials. In the present study, we first generated a binary tetracycline-on (Tet-On) system based on two lentivirus vectors, one expressing both human glial cell line-derived neurotrophic factor (hGDNF) and humanized recombinant green fluorescent protein (hrGFP) genes under second-generation tetracycline response element (TRE), and the other expressing the advanced reverse tetracycline-controlled transactivator--rtTA2S-M2 under a human minimal cytomegalovirus immediate early (CMV-IE) promoter. This system allows simultaneous expression of hGDNF and hrGFP genes in the presence of doxycycline (Dox). Human bone marrow-derived mesenchymal stem cells (hMSCs) were transduced with the binary Tet-On lentivirus vectors and characterized in vitro in the presence (On) or absence (Off) of Dox. The expression of hGDNF and hrGFP transgenes in transduced hMSCs was tightly regulated as determined by flow cytometry (FCM), GDNF enzyme-linked immunosorbent assay (ELISA) and quantitative real time-polymerase chain reaction (qRT-PCR). There was a dose-dependent regulation for hrGFP transgene expression. The levels of hGDNF protein in culture medium were correlated with the mean fluorescence intensity (MFI) units of hrGFP. The levels of transgene background expression were very low in the absence of Dox. The treatment of the conditioned medium from cultures of transduced hMSCs in the presence of Dox protected SH-SY5Y cells against 6-hydroxydopamine (6-OHDA) toxicity as determined by cell viability using 3, [4,5-dimethylthiazol-2-yl]-diphenyltetrazolium bromide (MTT) assay. The treatment of the conditioned medium was also found to improve the survival of dopaminergic (DA) neurons of ventral mesencephalic (VM) tissue in serum-free culture conditions as assessed by cell body area, the number of neurites and dendrite branching points, and proportion of tyrosine hydroxylase (TH)-immunoreactive (IR) cells. Our inducible lentivirus-mediated hGDNF gene delivery system may provide useful tools for basic research on gene therapy for chronic neurological disorders such as Parkinson's disease (PD).


Asunto(s)
Células de la Médula Ósea/metabolismo , Dopamina/metabolismo , Factor Neurotrófico Derivado de la Línea Celular Glial/genética , Lentivirus/genética , Células Madre Mesenquimatosas/metabolismo , Fármacos Neuroprotectores , Diferenciación Celular , Medios de Cultivo Condicionados , Doxiciclina/administración & dosificación , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Vectores Genéticos , Humanos , Técnicas In Vitro , Células Madre Mesenquimatosas/citología , Reacción en Cadena en Tiempo Real de la Polimerasa , Transgenes
4.
PLoS One ; 7(7): e41226, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-22815976

RESUMEN

Human albumin has recently been demonstrated to protect brain neurons from injury in rat ischemic brain. However, there is no information available about whether human albumin can prevent loss of tyrosine hydroxylase (TH) expression of dopaminergic (DA) neurons induced by 6-hydroxydopamine (6-OHDA) toxicity that is most commonly used to create a rat model of Parkinson's disease (PD). In the present study, two microliters of 1.25% human albumin were stereotaxically injected into the right striatum of rats one day before or 7 days after the 6-OHDA lesion in the same side. D-Amphetamine-induced rotational asymmetry was measured 7 days, 3 and 10 weeks after 6-OHDA lesion. We observed that intrastriatal administration of human albumin significantly reduced the degree of rotational asymmetry. The number of TH-immunoreactive neurons present in the substantia nigra was greater in 6-OHDA lesioned rats following human albumin-treatment than non-human albumin treatment. TH-immunoreactivity in the 6-OHDA-lesioned striatum was also significantly increased in the human albumin-treated rats. To examine the mechanisms underlying the effects of human albumin, we challenged PC12 cells with 6-OHDA as an in vitro model of PD. Incubation with human albumin prevented 6-OHDA-induced reduction of cell viability in PC12 cell cultures, as measured by MTT assay. Furthermore, human albumin reduced 6-OHDA-induced formation of reactive oxygen species (ROS) and apoptosis in cultured PC12 cells, as assessed by flow cytometry. Western blot analysis showed that human albumin inhibited 6-OHDA-induced activation of JNK, c-Jun, ERK, and p38 mitogen-activated protein kinases (MAPK) signaling in PC12 cultures challenged with 6-OHDA. Human albumin may protect against 6-OHDA toxicity by influencing MAPK pathway followed by anti-ROS formation and anti-apoptosis.


Asunto(s)
Albúminas/química , Oxidopamina/farmacología , Enfermedad de Parkinson/metabolismo , Tirosina 3-Monooxigenasa/metabolismo , Animales , Caspasa 3/metabolismo , Línea Celular Tumoral , Dextroanfetamina/farmacología , Modelos Animales de Enfermedad , Neuronas Dopaminérgicas/metabolismo , Femenino , Regulación Enzimológica de la Expresión Génica , Humanos , Técnicas In Vitro , Neuronas/metabolismo , Ratas , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno , Factores de Tiempo
5.
Brain Res ; 1327: 91-102, 2010 Apr 23.
Artículo en Inglés | MEDLINE | ID: mdl-20171958

RESUMEN

Convincing evidence has shown that brain ischemia causes the proliferation of neural stem cells/neural progenitor cells (NSCs/NPCs) in both the subventricular zone (SVZ) and the subgranular zone (SGZ) of adult brain. The role of brain ischemia-induced NSC/NPC proliferation, however, has remained unclear. Here we have determined whether brain ischemia-induced amplification of the NSCs/NPCs in adult brain is required for brain self-protection. The approach of intracerebroventricular (ICV) infusion of cytosine arabinoside (Ara-C), an inhibitor for cell proliferation, for the first 7days after brain ischemia was used to block ischemia-induced NSC/NPC proliferation. We observed that ICV infusion of Ara-C caused a complete blockade of NSC/NPC proliferation in the SVZ and a dramatic reduction of NSC/NPC proliferation in the SGZ. Additionally, as a result of the inhibition of ischemia-induced NSC/NPC pool amplification, the number of neurons in the hippocampal CA1 and CA3 was significantly reduced, the infarction size was significantly enlarged, and neurological deficits were significantly worsened after focal brain ischemia. We also found that an NSC/NPC-conditioned medium showed neuroprotective effects in vitro and that adult NSC/NPC-released brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF) are required for NSC/NPC-conditioned medium-induced neuroprotection. These data suggest that NSC/NPC-generated trophic factors are neuroprotective and that brain ischemia-triggered NSC/NPC proliferation is crucial for brain protection. This study provides insights into the contribution of endogenous NSCs/NPCs to brain self-protection in adult brain after ischemia injury.


Asunto(s)
Células Madre Adultas/fisiología , Infarto Encefálico/prevención & control , Isquemia Encefálica/patología , Ventrículos Laterales/patología , Neuronas/fisiología , Células Madre Adultas/efectos de los fármacos , Análisis de Varianza , Animales , Conducta Animal , Infarto Encefálico/etiología , Isquemia Encefálica/complicaciones , Bromodesoxiuridina/metabolismo , Células Cultivadas , Corteza Cerebral/citología , Medios de Cultivo Condicionados/farmacología , Citarabina/farmacología , Modelos Animales de Enfermedad , Proteínas Fluorescentes Verdes/genética , Inmunosupresores/farmacología , Etiquetado Corte-Fin in Situ/métodos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Proteínas del Tejido Nervioso/metabolismo , Examen Neurológico/métodos , Neuronas/efectos de los fármacos , Factores de Transcripción SOXB1/metabolismo
6.
J Cereb Blood Flow Metab ; 29(4): 759-70, 2009 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-19209180

RESUMEN

Chronic stroke is a highly important but under-investigated scientific problem in neurologic research. We have reported earlier that stem cell factor (SCF) in combination with granulocyte-colony stimulating factor (G-CSF) treatment during chronic stroke improves functional outcomes. Here we have determined the contribution of bone marrow-derived cells in angiogenesis and neurogenesis, which are enhanced by SCF+G-CSF treatment during chronic stroke. Using bone marrow tracking, flow cytometry, 2-photon live brain imaging, and immunohistochemistry, we observed that the levels of circulating bone marrow stem cells (BMSCs) (CD34+/c-kit+) were significantly increased by SCF+G-CSF treatment. In addition, live brain imaging revealed that numerous bone marrow-derived cells migrate into the brain parenchyma in the treated mice. We also found that bone marrow-derived cells, bone marrow-derived endothelial cells, vascular density, and bone marrow-derived neurons were significantly augmented by SCF+G-CSF. It is interesting that, in addition to the increase in bone marrow-derived endothelial cells, the number of bone marrow-derived pericytes was reduced after SCF+G-CSF treatment during chronic stroke. These data suggest that SCF+G-CSF treatment can enhance repair of brain damage during chronic stroke by mobilizing BMSCs, and promoting the contribution of bone marrow-derived cells to angiogenesis and neurogenesis.


Asunto(s)
Encéfalo/fisiología , Factor Estimulante de Colonias de Granulocitos/fisiología , Neurogénesis/fisiología , Regeneración/fisiología , Factor de Células Madre/fisiología , Accidente Cerebrovascular/patología , Animales , Células de la Médula Ósea , Encéfalo/patología , Circulación Cerebrovascular , Enfermedad Crónica , Ratones , Neovascularización Fisiológica , Células Madre/fisiología
7.
Zhonghua Er Ke Za Zhi ; 42(9): 659-62, 2004 Sep.
Artículo en Chino | MEDLINE | ID: mdl-15482665

RESUMEN

BACKGROUND: Corticosteroid treatment of acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) has been one of the most controversial clinical issues in critical care. Although the administration of high-dose corticosteroids does not benefit patients with early septic shock and ARDS, recent clinical trials have indicated that treatment with relatively low-dose corticosteroids (2 to 3 mg/kg/day of methylprednisolone or equivalent) may improve outcome when used for late ARDS or persistent septic shock. The underlying mechanism was not fully clarified. Whether the administration of corticosteroids can arrest neutrophil-driven organ injury once started remains to be elucidated. OBJECTIVE: To observe the effects of hydrocortisone (HC, 6 mg/kg) on oxygen free radicals (OFR) released by PMN and pulmonary pathological changes in rat ALI model induced by lipopolysaccharide (LPS), to investigate the possible mechanism through which corticosteroids exert protective effect on ALI. METHODS: A rat model of ALI was induced by peritoneal injection of 2 x 10(12) Escherichia coli/kg. Fifty-six rats were randomly divided into three groups: normal control group, LPS group and HC group (6 mg/kg). Samples were collected 2 h, 4 h and 6 h after giving LPS to LPS and HC group (6 h after giving normal saline in normal control group) to measure the level of OFR released by PMN using chemiluminescence method based on lumino, and to compae of pulmonary pathological changes among the three groups. RESULTS: Pathological examination with light microscope in LPS group showed thickened pulmonary interstitia, inflammatory cell infiltration, edema and hemorrhage, which were in accordance with the features of ALI. There were significant differences in the release of OFR by PMN among the three groups (P < 0.01). The level of OFR released by PMN in LPS group was significantly higher than that of the control group, and continued to increase during the observation period (2 - 6 h after LPS). The release of OFR by PMN in HC group was significantly suppressed as compared with LPS group, which was peaked at 4 h after LPS injection (to 98.2%); there were also significant differences in the grades of ALI pathologic changes among the three groups (P < 0.01). The grades of ALI pathologic changes in LPS group were significantly increased when compared with the normal control group (P < 0.05) while significantly decreased in HC group as compared with LPS group (P < 0.05). CONCLUSION: It was demonstrated in the LPS induced ALI model that OFR might play an important role in onset of ALI. Intervening with HC (6 mg/kg) treatment could ameliorate the lung injury and exert significant and sustained suppression on the release of OFR by PMN, showing that HC has a protective effect on LPS induced ALI and its theraputic effect occurs possibly through suppression on the release of OFR by PMN.


Asunto(s)
Lesión Pulmonar Aguda/inmunología , Radicales Libres/metabolismo , Glucocorticoides/farmacología , Hidrocortisona/farmacología , Lipopolisacáridos/efectos adversos , Neutrófilos/efectos de los fármacos , Lesión Pulmonar Aguda/etiología , Animales , Modelos Animales de Enfermedad , Pulmón/inmunología , Pulmón/patología , Ratones , Neutrófilos/metabolismo
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