Role of Cyclopentenone prostaglandins in promoting recovery after TBI

环戊烯酮前列腺素在促进 TBI 后恢复中的作用

• 批准号: 7870767
• 负责人: STEVEN H GRAHAM
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7870767
• 关键词: 9-deoxy-delta-9-prostaglandin D2; Acute; Agonist; Amyloid beta-Protein Precursor; Antidiabetic Drugs; Behavior; Behavioral; Binding; Brain; Capsid Proteins; Cell Death; Chimeric Proteins; Clinical; Clinical Trials; Cysteine; Data; Daughter; Dose; Enzyme-Linked Immunosorbent Assay; Enzymes; FDA approved; Fatty Acids; Functional disorder; Gelatinase B; Glutathione; HIV; Healed; Hydrolase; Hypoxia; Immunoblotting; Immunohistochemistry; In Situ Nick-End Labeling; Infection; Inflammation; Injury; Ions; Lead; Ligands; Measures; Mediating; Metabolism; Modeling; Monitor; Mus; Mutation; Neurons; Nuclear Receptors; Outcome; Parents; Pathway interactions; Patients; Peroxidases; Peroxisome Proliferator-Activated Receptors; Play; Population; Procedures; Production; Prostaglandin-Endoperoxide Synthase; Prostaglandins; Proteins; Prothrombin; Rattus; Reaction; Recombinants; Recovery; Reporting; Resolution; Rodent Model; Role; Short-Term Memory; TBI Patients; Testing; Time; Toxic effect; Translating; Trauma; Traumatic Brain Injury; UCHL1 gene; Ubiquitin; Variant; Veterans; Wound Healing; brain tissue; controlled cortical impact; cyclopentenone; effective therapy; enzyme activity; enzyme pathway; healing; improved; in vivo; injured; insight; migration; morris water maze; multicatalytic endopeptidase complex; neuron loss; neutrophil; novel; novel strategies; novel therapeutic intervention; prevent; receptor; research study; restoration; rosiglitazone; ubiquitin C-terminal hydrolase

DESCRIPTION (provided by applicant): Traumatic brain injury (TBI) is a major problem in the veteran population and there are no effective treatments proven to improve long-term recovery. In new preliminary data, we have found that the concentration of the cyclopentenone prostaglandin (CyPG) D12-prostaglandin J2 (D12-PGJ2) is increased in rat brain after TBI. CyPGs such as D12-PGJ2 are potent ligands of the PPARg nuclear receptor, and have been proposed to play an important role in resolution of inflammation and wound healing after injury and infection. CyPGs may also injure neurons by non-PPAR3-mediated effects including disruption of the ubiquitin proteasome pathway (UPP). In additional preliminary data, we have found that CyPGs bind to and inhibit the action of the key neuronal UPP enzyme, ubiquitin carboxy-terminal hydrolase-L1 (UCH-L1) in neurons resulting in accumulation of ubiquitinated proteins and neuronal cell death. In these proposed studies, we will further characterize the production of and metabolism of CyPGs after TBI. We will also determine the role of PPARg activation and restoration of UCH-L1 activity in determining recovery after TBI and long term behavioral outcome. The following specific aims are proposed: 1. Characterize the time course of production of cyclopentenone prostaglandins and their metabolites after controlled cortical injury (CCI) in rats. 2. Test whether activation of the PPAR3 receptors reduces secondary injury and improves behavioral recovery after TBI. 3. Test whether restoring UCH-L1 activity by systemic treatment with a TAT-UCH-L1 fusion protein can prevent accumulation of Ub-proteins and improve behavioral outcome after TBI. TBI will be induced using the controlled cortical impact injury model in rats and mice. Concentrations of CyPGs will be measured in brain after TBI by quadrupole LC MS/MS using selective reaction monitoring of daughter ion fragments of the CyPG parent masses. Neutrophil migration will be quantified by measuring myleoperoxidase activity in brain tissue. The proteins, iNOS, TNFa, and MMP-9, which are known to be regulated by PPARg, will be measured by ELISA and used as downstream monitors of PPARg activation. Disruption of the UPP in brain after TBI will be determined by monitoring UCH-L1 activity and detecting accumulation of ubquitinated proteins by immunoblotting and immunohistochemistry. Behavioral outcome will be assessed using a working memory variation of the Morris Water Maze procedure assessed on post trauma days 14 through 20. These preliminary data represent the first report that CyPGs are present in brain after TBI. Activation of PPARg and restoration of lost UCH-L1 activity are new therapeutic approaches to TBI that could lead to novel therapies that could improve long term recovery and restoration of function after TBI. PUBLIC HEALTH RELEVANCE: Traumatic brain injury (TBI) is a major problem in the veteran population and no treatments have yet been proven to improve long-term recovery. The purpose of this proposal is to test novel therapeutic approaches in rodent models of TBI that could lead to new approaches to improved recovery after TBI in patients. In new preliminary data, we have found that fatty acids, known as cyclopentenone prostaglandins, are increased in rat brain after TBI. These fatty acids can activate PPARg, a nuclear receptor that is the target of anti-diabetic drugs, and improve healing. However, we have also found that these fatty acids inhibit an important enzyme in neurons known as ubiquitin carboxy hydrolase-L1 (UCH-L1). In the proposed studies, we will test whether the FDA-approved anti-diabetic drug rosiglitazone, that targets the PPARg receptor, can improve behavioral outcome after TBI. We will also test whether treatment with a modified UCH-L1 protein can restore the lost UCH-L1 enzyme activity and improve behavioral outcome after TBI.

描述（由申请人提供）： 创伤性脑损伤（TBI）是退伍军人群体中的一个主要问题，并且没有有效的治疗方法被证明可以改善长期恢复。在新的初步数据中，我们已经发现，环戊烯酮前列腺素（CyPG）D12-前列腺素J2（D12-PGJ 2）的浓度在TBI后大鼠脑中增加。CyPG如D12-PGJ 2是PPARg核受体的有效配体，并且已经提出在损伤和感染后的炎症消退和伤口愈合中发挥重要作用。CyPG也可能通过非PPAR 3介导的作用损伤神经元，包括破坏泛素蛋白酶体途径（UPP）。在额外的初步数据中，我们发现CyPG结合并抑制神经元中关键的神经元UPP酶，泛素羧基末端水解酶-L1（UCH-L1）的作用，导致泛素化蛋白的积累和神经元细胞死亡。 在这些拟议的研究中，我们将进一步表征TBI后CyPG的产生和代谢。我们还将确定PPARg激活和UCH-L1活性恢复在确定TBI后恢复和长期行为结果中的作用。提出了以下具体目标：1。描述大鼠可控性皮质损伤（CCI）后环戊烯酮野牡丹素及其代谢产物产生的时间过程。2.测试PPAR 3受体的激活是否减少TBI后的继发性损伤并改善行为恢复。3.测试通过用TAT-UCH-L1融合蛋白进行全身治疗来恢复UCH-L1活性是否可以防止Ub蛋白的积累并改善TBI后的行为结果。 将使用大鼠和小鼠中的受控皮质撞击损伤模型诱导TBI。TBI后，通过四极杆LC MS/MS，使用CyPG母体分子的子离子片段的选择性反应监测，测量脑中的CyPG浓度。将通过测量脑组织中的髓过氧化物酶活性来定量神经元迁移。已知由PPARg调节的蛋白质iNOS、TNF α和MMP-9将通过ELISA测量并用作PPARg活化的下游监测物。通过监测UCH-L1活性并通过免疫印迹和免疫组织化学检测泛素化蛋白的积累来确定TBI后脑中UPP的破坏。将使用在创伤后第14天至第20天评估的Morris水迷宫程序的工作记忆变化来评估行为结果。 这些初步数据代表了TBI后脑中存在CyPG的首次报道。PPARg的激活和失去的UCH-L1活性的恢复是TBI的新治疗方法，其可以导致可以改善TBI后的长期恢复和功能恢复的新疗法。 公共卫生相关性： 创伤性脑损伤（TBI）是退伍军人群体的一个主要问题，目前还没有任何治疗方法被证明可以改善长期恢复。该提案的目的是在TBI的啮齿动物模型中测试新的治疗方法，这些方法可能导致改善患者TBI后恢复的新方法。在新的初步数据中，我们发现，脂肪酸，称为环戊烯酮类化合物，在TBI后大鼠大脑中增加。这些脂肪酸可以激活PPARg，一种抗糖尿病药物靶向的核受体，并改善愈合。然而，我们还发现这些脂肪酸抑制神经元中的一种重要酶，称为泛素羧基水解酶-L1（UCH-L1）。在拟议的研究中，我们将测试FDA批准的靶向PPARg受体的抗糖尿病药物罗格列酮是否可以改善TBI后的行为结果。我们还将测试用修饰的UCH-L1蛋白治疗是否可以恢复失去的UCH-L1酶活性并改善TBI后的行为结果。