NEUROPROTECTIVE MECHANISMS OF STATINS IN NEURONS

他汀类药物对神经元的神经保护机制

• 批准号: 7192132
• 负责人: WELLINGTON GIBSON WOOD
• 金额: $ 20.16万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-12-01 至 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7192132
• 关键词: 3-hydroxy-3-methylglutaryl-coenzyme A; AIDS Dementia Complex; Alzheimer' s Disease; Amyloid; Amyloid beta-Protein Precursor; Animals; Antioxidants; Apoptosis; Apoptotic; Astrocytes; BCL2 gene; Binding; Blood Platelets; Blood Vessels; Calcineurin; Cardiac Myocytes; Cell Death; Cell Nucleus; Cell Survival; Cell model; Cell physiology; Cells; Cerebral cortex; Characteristics; Cholesterol; Class; Clinical Trials; Collaborations; Complement 3; Cyclodextrins; Data; EMSA; Endothelin-1; Family; Family member; GTP-Binding Proteins; Gene Expression; Genes; HA14-1; Hippocampus (Brain); Human; In Vitro; Indiana; Inflammation; Inflammatory; Inflammatory Response; Ischemia; Ischemic Stroke; Lead; Lipids; Lovastatin; Membrane; Messenger RNA; Metabolism; Mitochondria; Multiple Sclerosis; Mus; N-Methylaspartate; NADPH Oxidase; Neuroblastoma; Neurodegenerative Disorders; Neuroglia; Neurons; Nuclear; Nucleotides; Oblimersen; Oryctolagus cuniculus; Oxidative Stress; Oxidoreductase; P2Y2 receptor; Paper; Pathway interactions; Patients; Personal Satisfaction; Pharmaceutical Preparations; Physiological reperfusion; Play; Pravastatin; Preparation; Process; Production; Promoter Regions; Protein Biosynthesis; Protein Family; Protein Isoprenylation; Protein Overexpression; Proteins; Protocols documentation; Regulation; Reperfusion Therapy; Reporting; Research Personnel; Resources; Role; Signal Transduction; Simvastatin; Site; Small Interfering RNA; Smooth Muscle Myocytes; Sterols; T-Lymphocyte; Testing; Thinking; Time; Toxic effect; Transcriptional Activation; Transcriptional Regulation; Transgenic Mice; Up-Regulation; Wood material; apoptosis inducing factor; astrogliosis; atorvastatin; brain tissue; cell growth; cell type; excitotoxicity; farnesyl pyrophosphate; farnesylation; geranylgeranyl pyrophosphate; geranylgeranylation; hypercholesterolemia; in vivo; inhibitor/antagonist; innovation; insight; interest; isoprenoid; isoprenylation; member; mevalonate; mitochondrial dysfunction; mutant; neuron apoptosis; neuroprotection; novel; prenylation; prevent; programs; research study; rho; rho GTP-Binding Proteins; trafficking; transcription factor

Our focus is on cholesterol-independent and dependent mechanisms whereby statins afford neuroprotection to cells undergoing oxidative stress. Statins reduce cholesterol levels but also they have cholesterol- independent mechanisms of action. Neuroprotective effects of statins could involve multiple pathways.We have discovered that statins altered expression of genes involved in apoptosis, cell growth, signaling and trafficking in the murine cerebral cortex. A novel finding was that simvastatin increased gene expressionand protein levels of Bcl-2 in vivo and in vitro. Bcl-2 and anti-apoptotic members of the Bcl-2 family play apivotal role in neuronal cell survival. Simvastatin also significantly increased ET-1 expression levels whose product is the precursor for the ET-1 protein. ET-1 is involved in transcriptional activation of Bcl-2. Preliminary data revealed that when Bcl-2 protein levels were experimentally reduced the neuroprotective effects of simvastatin to an Ap challenge were eliminated. We hypothesizethat: Simvastatin has both cholesterol- independent and-dependent mechanisms of action that are neuroprotective. Neuroprotective mechanisms are due to regulation of apoptosis by Bcl-2 family members, including the transcriptional regulation of Bcl-2 by ET-1/calcineurin/NFAT-dependent pathways and inhibition of Rac1 geranylgeranylation. These hypotheses will be testing primarily in mouse primary cortical and hippocampal neurons. Aim 1. Determine Bcl-2 and Bax gene expression and protein levels in neurons pretreated with simvastatin and challenged by A(Jor NMDA. Determine if simvastatin-induced increasein Bcl-2 inhibits translocation of apoptosis -inducing factor induced by Ap or NMDA from the mitochondria to the nucleus in neurons. Examine whether suppression of Bcl-2 gene expression or inactivation of the protein diminishes neuroprotective effects of simvastatin in neuroblastoma cells treated with Ap or NMDA. Aim 2. Determine if ET-1 increases Bcl-2 gene expression and protein levels in neurons challenged with Ap or NMDA. Evaluate if ET-1/calcineurin induces NFAT to the nucleus, binding to the NFAT sites on the Bcl-2 promoter region and increases Bcl-2 gene expression in neurons. Determine if suppression of ET-1 expression reduces simvastatin-induced stimulation of Bcl-2 gene expression and protein levels and neuroprotection when challenged by Ap or NMDA in human neuroblastoma cells. Aim 3. Determine if lowering cholesterol levels directly by cyclodextrin increases Bcl-2 and ET-1 gene expression andprotein levels in neurons and if neuroprotective. Evaluate if Bcl-2 and ET-1 gene expression and protein levels and neuroprotection are increased when farnesylation and geranylgeranylation are inhibited by non-statin inhibitors in neurons. Determine if simvastatin-induced stimulation of Bcl-2 and ET-1 gene expression and protein levels and neuroprotection can be inhibited by isoprenoids in neurons.

我们的重点是胆固醇的非依赖性和依赖性机制，他汀类药物提供神经保护 氧化应激的细胞。他汀类药物降低胆固醇水平，但他们也有胆固醇- 独立的行动机制。他汀类药物的神经保护作用可能涉及多个途径。 他们发现他汀类药物改变了参与细胞凋亡、细胞生长、信号传导和 老鼠大脑皮层的运输一个新的发现是辛伐他汀增加了基因表达， Bcl-2蛋白水平在体内和体外。Bcl-2和Bcl-2家族的抗凋亡成员在细胞凋亡中起着至关重要的作用。 在神经细胞存活中的作用辛伐他汀还显著增加ET-1表达水平，其产物 是ET-1蛋白的前体ET-1参与Bcl-2的转录激活。初步数据 揭示了当Bcl-2蛋白水平在实验上降低时， 辛伐他汀对Ap激发的抑制作用被消除。我们假设：辛伐他汀既有胆固醇- 神经保护作用的独立和依赖机制。神经保护 机制是由于Bcl-2家族成员对凋亡的调节，包括 通过ET-1/钙调神经磷酸酶/NFAT依赖性途径调节Bcl-2的转录， Rac 1香叶基香叶基化。这些假设将主要在小鼠初级皮质和 海马神经元目标1.测定神经元Bcl-2和Bax基因表达及蛋白水平 用辛伐他汀预处理并用A或NMDA攻击。确定辛伐他汀诱导的 Bcl-2抑制Ap或NMDA诱导的凋亡诱导因子从线粒体转位到线粒体 神经元的细胞核。检查Bcl-2基因表达的抑制或蛋白质的失活 降低辛伐他汀在用Ap或NMDA处理的神经母细胞瘤细胞中的神经保护作用。目标2. 确定ET-1是否增加Ap或Ap刺激的神经元中Bcl-2基因表达和蛋白水平， NMDA评估ET-1/钙调神经磷酸酶是否诱导NFAT进入细胞核，与Bcl-2上的NFAT位点结合 启动子区和增加Bcl-2基因表达的神经元。确定ET-1是否受到抑制 表达降低辛伐他汀诱导的Bcl-2基因表达和蛋白水平的刺激， 在人神经母细胞瘤细胞中，当被Ap或NMDA攻击时的神经保护作用。目标3.确定是否 通过环糊精直接降低胆固醇水平增加Bcl-2和ET-1基因表达和蛋白 在神经元中的水平以及是否具有神经保护作用。评估Bcl-2和ET-1基因表达和蛋白水平， 当法尼基化和香叶基香叶基化被非他汀抑制时， 神经元中的抑制剂。确定辛伐他汀是否诱导Bcl-2和ET-1基因表达的刺激， 蛋白质水平和神经保护可被神经元中的类异戊二烯抑制。