Cord Blood is Neuroprotective in a Rat Model of Stroke

脐带血对中风大鼠模型具有神经保护作用

• 批准号: 7585234
• 负责人: ALISON E WILLING
• 金额: $ 34.48万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-15 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7585234
• 关键词: Address; Adverse effects; Affect; Aftercare; Age; Alteplase; Animal Model; Anti-Inflammatory Agents; Anti-inflammatory; Apoptosis; Apoptotic; Astrocytes; B-Lymphocytes; Behavioral; Binding; Biological Assay; Blood Cells; Brain; Brain Injuries; Cell Culture Techniques; Cell Death; Cell Fraction; Cell Separation; Cell Therapy; Cell Transplantation; Cells; Cerebrovascular Disorders; Cessation of life; Chloride Ion; Chlorides; Clinical; DNA Binding; Disease; Flow Cytometry; Gene Expression; Genes; Genetic; Goals; Hour; Human; Hypoxia; Immune; Immunohistochemistry; In Situ Nick-End Labeling; In Vitro; Infarction; Infiltration; Inflammation; Inflammatory Response; Inflammatory Response Pathway; Injection of therapeutic agent; Injury; Label; Measures; Mediating; Microarray Analysis; Microglia; Middle Cerebral Artery Occlusion; Modeling; Mononuclear; Morbidity - disease rate; Motor; NF-kappa B; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Oligodendroglia; Outcome Measure; Pathology; Performance; Peripheral; Population; Population Heterogeneity; Rattus; Recovery; Reporting; Research; Series; Signal Transduction; Site; Staining method; Stains; Stem cells; Stroke; T-Lymphocyte; Testing; Tetrazolium; Time; Transplantation; Umbilical Cord Blood; Umbilical cord structure; United States; Western Blotting; blood treatment; cell type; cytokine; disability; effective therapy; improved; inflammatory marker; innovation; monocyte; mortality; neuronal survival; neuroprotection; programs; relating to nervous system; repaired; research study; response; stroke recovery; therapy development

Cerebrovascular disease is the third leading cause of morbidity and mortality in the United States. As the population ages, it will be even more pressing to develop effective treatment options that can not only increase survival, but decrease disability. In a rat model of stroke (the middle cerebral artery occlusion or MCAO) we found that intravenously (iv) administering the mononuclear fraction from human umbilical cord blood (HUCB) enhances motor functions. In this proposal we will explore how the administration of this population of cells may induce recovery and decrease anatomical damage induced by MCAO through direct neuroprotective and anti-inflammatory mechanisms. In Aim 1we will characterize how iv HUCB cells modify the underlying pathology and inflammation of the MCAO when administered 24 hours to 7 days after the MCAO using markers of inflammation, neuronal death, neurosurvival, and apoptosis. In Aim 2, we will identify the specific subpopulation of the HUCB cells that is instrumental in inducing behavioral and anatomical recovery. This will be accomplished using fluorescent activated cell sorting (FACS) to enrich T cell, B Cell, monocyte and "stem" cell fractions for transplantation into a stroked rat. Endpoint measures in these studies will include performance on a battery of motor tests and infarct volume. Aim 3 proposes a series of in vitro studies to examine whether the HUCB cells modify the inflammatory response to the MCAO through direct interactions with neural cell popultions. Cell culture assays using neuronal, astrocytic, oligodendrocytes or microglia will be conducted to identify direct interactions and the molecular signals that mediate survival, cytokine expression and NF-kappaB binding activity in response to hypoxia/reoxygenation. In Aim 4 we will use microarray technology to identify neural repair/survival genes that are up-regulated by HUCB treatment of neuronal cultures. All microarray experiments will be verified with western blots and immunohistochemistry. The studies proposed here will increase our understanding of the neuroprotective and anti-inflammatory mechanisms underlying the recovery from stroke induced by HUCB cell transplantation and may also identify other potential targets in stroke for development of treatment options. The ultimate goal of this research program is to develop an innovative HUCB cell based therapy into a viable clinical option for transplantation in neurodegenerative disease and brain injury.

脑血管疾病是美国发病率和死亡率的第三大原因。为 随着人口老龄化，开发有效的治疗方案将更加紧迫， 提高存活率，但减少残疾。在大鼠中风模型（大脑中动脉闭塞或 MCAO），我们发现静脉内（iv）给予人脐带单个核组分 血液（HUCB）增强运动功能。在本建议中，我们将探讨如何管理这一 细胞群可以通过直接的细胞毒性诱导恢复并减少由MCAO诱导的解剖损伤。 神经保护和抗炎机制。在目的1中，我们将描述静脉注射HUCB细胞如何 给药后24小时至7天，改变MCAO的潜在病理和炎症 MCAO使用炎症、神经元死亡、神经存活和细胞凋亡的标志物。在目标2中，我们将 鉴定有助于诱导行为和免疫反应的HUCB细胞的特定亚群， 解剖恢复这将使用荧光激活细胞分选（FACS）来富集T细胞， 细胞、B细胞、单核细胞和“干”细胞组分用于移植到中风大鼠中。终点测量 这些研究将包括一系列运动测试和梗塞体积的表现。目标3提出了一个 一系列体外研究，以检查HUCB细胞是否改变对MCAO的炎症反应 通过与神经细胞群的直接相互作用。使用神经元，星形胶质细胞， 将进行少突胶质细胞或小胶质细胞，以鉴定直接相互作用和分子信号， 介导存活、细胞因子表达和NF-κ B结合活性。 在目标4中，我们将使用微阵列技术来鉴定神经修复/存活基因，这些基因是由以下因素上调的： 神经元培养物的HUCB处理。所有的微阵列实验将用蛋白质印迹进行验证， 免疫组化这里提出的研究将增加我们对神经保护作用的理解。 HUCB细胞诱导脑卒中恢复的抗炎机制 此外，还可以确定中风中的其他潜在靶点，以开发治疗方案。 这项研究计划的最终目标是开发一种创新的基于HUCB细胞的治疗方法， 在神经退行性疾病和脑损伤中移植的可行临床选择。