RESUMO
When human umbilical cord blood cells (HUCBCs) are administered intravenously after a middle cerebral artery occlusion, they reliably produce behavioral and anatomical recovery, and protect neural tissue from progressive change. However, our results indicate that the cells do not exert their effects by engraftment in the peri-infarct region, even though they migrate to the site of injury. The objective of the present study was to determine if the cells induce recovery by decreasing inflammation. We used a combination of in vivo and in vitro studies to show that HUCBCs decrease inflammation in the brain after stroke and thereby enhance neuroprotection. After stroke and transplantation, there was a decrease in CD45/CD11b- and CD45/B220-positive (+) cells. This decrease was accompanied by a decrease in mRNA and protein expression of pro-inflammatory cytokines and a decrease in nuclear factor kappaB (NF-kappaB) DNA binding activity in the brain of stroke animals treated with HUCBCs. In addition to modulating the inflammatory response, we demonstrate that the cord blood cells increase neuronal survival through non-immune mechanisms. Once thought of as "cell replacement therapy," we now propose that cord blood treatment in stroke reduces inflammation and provides neuroprotection. Both of these components are necessary for effective therapy.
Assuntos
Transplante de Células , Sangue Fetal/citologia , Inflamação/metabolismo , Acidente Vascular Cerebral/terapia , Animais , Encéfalo/citologia , Encéfalo/metabolismo , Antígeno CD11b/imunologia , Separação Celular , Sobrevivência Celular , Citocinas/imunologia , Modelos Animais de Doenças , Citometria de Fluxo , Humanos , Infarto da Artéria Cerebral Média , Antígenos Comuns de Leucócito/imunologia , Masculino , NF-kappa B/metabolismo , Neurônios/citologia , Neurônios/metabolismo , Ratos , Ratos Sprague-Dawley , Acidente Vascular Cerebral/imunologia , Acidente Vascular Cerebral/patologiaRESUMO
Animal models of cerebral infarction are crucial to understanding the mechanisms of neuronal survival following ischemic brain injury and to the development of therapeutic interventions for victims of all types of stroke. Rodents have been used extensively in such research. One rodent model of stroke utilizes either permanent or temporary occlusion of the middle cerebral artery (MCAO) to produce ischemia. Since the development of an endovascular method for this was published in 1989, MCAO has been applied commonly to the rat, and often paired with 2, 3, 5-triphenyltetrazolium chloride (TTC) staining for stroke volume measurement. Meanwhile, advances in the ability to genetically alter mice have allowed exciting lines of research into ischemia. Because of technical demands and issues with survival, relatively few laboratories have investigated the MCAO method in the mouse. Our present work utilizes a mouse middle cerebral occlusion (MCAO) model of embolic stroke to study neuronal degeneration following temporary focal cerebral ischemia. C57Bl/6J mice were used to examine the exact effects of MCAO using Fluoro-Jade, a marker of neurodegeneration that allows observation of specific brain regions and cells destined to die. A time course of escalating neuronal degeneration from 10 min to 7 days following MCAO was established. Technical aspects of this popular method for transient focal ischemia as it applies to the mouse are discussed.
Assuntos
Encéfalo/patologia , Infarto da Artéria Cerebral Média/patologia , Ataque Isquêmico Transitório/patologia , Degeneração Neural/patologia , Neurônios/patologia , Animais , Biomarcadores/análise , Encéfalo/citologia , Lesões Encefálicas/patologia , Morte Celular , Córtex Cerebral/citologia , Córtex Cerebral/patologia , Corpo Estriado/citologia , Corpo Estriado/patologia , Modelos Animais de Doenças , Fluoresceínas , Corantes Fluorescentes/análise , Hipocampo/citologia , Hipocampo/patologia , Camundongos , Camundongos Endogâmicos C57BL , Compostos Orgânicos , Coloração e Rotulagem/métodos , Sais de Tetrazólio/análise , Fatores de TempoRESUMO
Activation of several immediate early genes (IEGs) is crucial for long-term memory formation in vivo. In vitro methods of inducing these genes have not been investigated extensively. Here we present data demonstrating that application of the neurotrophin, nerve growth factor (NGF), to both rat primary neuronal cultures and differentiated mouse neuroblastoma 2A (N2A) cultures reliably induces expression of several IEGs, including Zif268, Nur77 and Arc, each of which have been linked to memory consolidation. These findings provide an in vitro model in which to test other agents that might modulate the induction of memory-associated genes.