Effects of Long-Term Morphine Treatment on Opioid Receptor Signaling and Inflammation in the Chronic Post-TBI Period

长期吗啡治疗对慢性 TBI 后阿片受体信号传导和炎症的影响

• 批准号: 10454092
• 负责人: Kelly Bosse
• 金额: --
• 依托单位: JOHN D DINGELL VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10454092
• 关键词: Absence of pain sensation; Acute; Address; Affect; Affinity; Agonist; Amygdaloid structure; Analgesics; Area; Astrocytes; Award; Awareness; Behavior; Behavioral; Binding; Biological Assay; Brain; Brain Injuries; Cell Nucleus; Chronic; Clinical; Cognition; Cognitive; Coupled; Data; Dependence; Diagnosis; Dorsal; Dose; Emotional; Enzyme-Linked Immunosorbent Assay; Epidemic; Experimental Designs; Exposure to; Extracellular Signal Regulated Kinases; Female; Flow Cytometry; Foundations; Freedom; Functional disorder; Future; GTP-Binding Proteins; Gene Expression; Guanosine Triphosphate; Hippocampus (Brain); Human Resources; Hyperalgesia; Immunity; Immunofluorescence Immunologic; Impaired cognition; Impairment; Individual; Inflammation; Inflammatory; Inflammatory Response; Injury; Interleukin-1 beta; Label; Ligand Binding; Link; Long-Term Care; Long-Term Effects; Measurement; Measures; Mediating; Mediator of activation protein; Microglia; Mission; Molecular Analysis; Morphine; Mus; Narcotics; Nature; Neuroglia; Neurologic; Neurons; Nociception; Nucleus Accumbens; Opioid; Opioid Receptor; Outcome; Pain; Pain management; Paper; Pathologic; Pathology; Pathway interactions; Patient Care; Pattern; Pharmaceutical Preparations; Pharmacology; Phenotype; Population; Precipitation; Prefrontal Cortex; Process; Property; Proteins; Published Comment; Publishing; Receptor Signaling; Recurrent pain; Regimen; Rehabilitation therapy; Rewards; Risk; Services; Signal Pathway; Signal Transduction; Signaling Protein; Stains; Substance Use Disorder; System; TLR4 gene; Testing; Time; Tissues; Traumatic Brain Injury; Ventral Tegmental Area; Veterans; Withdrawal; addiction; adverse outcome; base; chronic pain management; cytokine; density; effective therapy; glial activation; high risk; immune activation; immune system function; improved; innovation; interest; male; midbrain central gray substance; morphine administration; mu opioid receptors; novel; operation; opioid misuse; opioid use; pain relief; radioligand; receptor; receptor density; receptor expression; recruit; response; service member; side effect; tool

Increasing awareness has been placed in understanding the issues related to rehabilitation of the estimated 330,000 service personnel diagnosed with traumatic brain injury (TBI) since the start of Operation Enduring Freedom and Operation Iraqi Freedom, who suffer from physical, cognitive, emotional dysfunction and recurrent pain. Specifically, opioid misuse is a growing epidemic nationwide, and recent evidence indicates that Veterans who sustain moderate/severe TBI are more likely to receive long-term treatment with opioid- based medications and engage in higher-risk opioid use patterns. These findings highlight the critical need for studies to establish what adaptive changes occur in the opioid system in the injured brain that may influence nociceptive, cognitive and reward/dependence outcomes. Morphine, the prototypical opioid antinociceptive, binds to the mu opioid receptor (MOR) to stimulate G-protein signaling and downstream effectors, such as extracellular signal-regulated kinase (ERK). In addition to this classical signaling pathway, morphine can act through the immunity toll-like like receptor 4 (TLR4) to activate glial cells and stimulate the release of proinflammatory mediators, like interleukin 1 beta (IL-1). While both signaling pathways contribute to morphine-induced phenotypes, evidence indicates immune activation disrupts morphine’s efficacy to promote analgesia, worsens pathological pain and exacerbates dependence and withdrawal. While TBI is recognized to induce long-term effects on immune system function alone, to date, no published papers have explored the combined influence of TBI and opioids on these signaling outcomes, adding to the significance and innovation of the proposed project. The current SPiRE proposal will test the hypothesis that repeated exposure to morphine following TBI will upregulate MOR expression/function and selectively potentiate glial-mediated inflammatory processes upon subsequent re-exposure to morphine in regions associated with reward, cognition and pain. The objectives in this proposal will be accomplished through two Specific Aims with tissue measurements in the nucleus accumbens, ventral tegmental area, prefrontal cortex, periaqueductal gray, dorsal hippocampus and amygdala. Aim 1 will assess longitudinal changes in MOR status as measured by radioligand binding density/affinity as well as gene expression (real-time PCR) and immunofluorescent co- localization on neurons and glia in tissue collected 7 days, 1, 3 and 6 months following TBI/repeated morphine. Aim 2 will characterize differential activation of classical and inflammatory pathways stimulated by morphine re- exposure at long-term timepoints (3 and 6 months) following combined TBI and subchronic morphine. Activity through classical pathways will be evaluated by G-protein (GTPS) activity and extent of phosphorylated ERK immunofluorescence. Inflammatory pathway signaling will be assessed by measuring the extent of glial activation with immunofluorescence, TLR4 microglial staining and flow cytometry, and IL-1 protein quantification with ELISA. The hypothesized finding of a potentiated inflammatory response to morphine following TBI would be consistent with conferring an increased risk of adverse outcomes associated with activation of this pathway, including addiction vulnerability, impaired cognition and hyperalgesia, making these studies highly relevant to the mission for VA patient care and rehabilitation. These data will also provide critical evidence to serve as a foundation for a comprehensive future study to explore the behavioral implications of these signaling outcomes and pharmacologic strategies exploring biased agonism.

人们越来越多地认识到与修复有关的问题 自行动开始以来，估计有330,000名服务人员被诊断为创伤性脑损伤(TBI) 持久的自由和伊拉克自由行动，他们遭受身体、认知和情感功能障碍的折磨 以及反复发作的疼痛。具体地说，阿片类药物滥用在全国范围内日益流行，最近的证据表明 患有中/重度脑外伤的退伍军人更有可能接受阿片类药物的长期治疗- 以药物为基础，从事风险较高的阿片类药物使用模式。这些发现突显了 研究确定在受损的大脑中阿片系统发生了什么适应性变化，可能会影响 伤害性、认知性和奖赏/依赖结果。吗啡，典型的阿片类抗伤害性感受剂， 结合Mu阿片受体(MOR)以刺激G蛋白信号和下游效应器，如 细胞外信号调节激酶(ERK)。除了这个经典的信号通路外，吗啡还可以起作用 通过免疫Toll样受体4(TLR4)激活神经胶质细胞并刺激其释放 促炎介质，如白介素1β(IL-1)。虽然这两条信号通路都有助于 吗啡诱导的表型，证据表明免疫激活扰乱了吗啡的促进作用 止痛，加重病理性疼痛，加剧依赖和戒断。虽然TBI被认为是 单独对免疫系统功能产生长期影响，到目前为止，还没有发表的论文探讨 脑损伤和阿片类药物对这些信号转导结果的联合影响，增加了意义和创新 所提议的项目。 目前的SPIRE提议将检验以下假设：脑外伤后反复接触吗啡 是否会上调MOR的表达/功能并选择性地增强胶质细胞介导的炎症 在随后再次接触吗啡时，与奖赏、认知和 疼痛。本提案中的目标将通过组织方面的两个具体目标来实现 伏隔核、腹侧被盖区、前额叶皮质、中脑导水管周围灰质、 背侧海马体和杏仁核。目标1将评估MOR状态的纵向变化，通过 放射配基结合密度/亲和力以及基因表达(实时聚合酶链式反应)和免疫荧光共标记 脑损伤后7d、1、3、6个月组织中神经元和神经胶质细胞的定位。 目的2描述吗啡再刺激的经典和炎症通路的不同激活。 在脑损伤合并亚慢性吗啡后的长期时间点(3个月和6个月)暴露。活动 通过经典通路将G蛋白(GTP、、S)的活性和磷酸化程度进行评估 免疫荧光。炎症通路信号将通过测量神经胶质细胞的程度来评估 免疫荧光激活、TLR4小胶质细胞染色和流式细胞术，以及IL-1蛋白 用ELISA法进行定量。对吗啡炎症反应增强的假想发现 脑外伤后，与以下相关的不良后果风险增加是一致的 这一通路的激活，包括成瘾易感性、认知受损和痛觉过敏，使这些 与退伍军人管理局患者护理和康复任务高度相关的研究。这些数据还将提供关键的 作为未来全面研究的基础的证据，以探索 这些信号结果和药物策略探索有偏见的激动症。