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.

越来越多的意识已成为理解与康复有关的问题 自手术开始以来，估计有330,000名服务人员被诊断出患有创伤性脑损伤（TBI） 持久的自由和行动伊拉克自由，患有身体，认知，情感功能障碍 和反复疼痛。具体而言，阿片类药物是全国流行病的日益增长，最近的证据表明 维持中等/重度TBI的退伍军人更有可能接受阿片类药物的长期治疗 基于药物并从事高风险的阿片类药物使用模式。这些发现突出了对 研究可能会影响受伤大脑中阿片类药物系统中发生哪些适应性变化的研究 伤害性，认知和奖励/依赖性结果。吗啡，原型阿片类药物抗伤害感受， 与MU阿片受体（MOR）结合以刺激G蛋白信号传导和下游效应，例如 细胞外信号调节激酶（ERK）。除了这种经典信号通路，吗啡还可以作用 通过像接收器4（TLR4）这样的免疫收费，以激活神经胶质细胞并刺激释放 促炎介质，例如白介素1β（IL-1）。而两种信号通路都有助于 吗啡诱导的表型，证据表明免疫激活破坏了吗啡的效率促进 镇痛，使病理疼痛恶化，并加剧依赖性和戒断。而TBI被认为 迄今为止，尚无公开的论文探讨了对免疫系统功能的长期影响 TBI和阿片类药物对这些信号结果的综合影响，增加了意义和创新 拟议的项目。 当前的尖峰提案将检验以下假设，即TBI后重复接触吗啡 将更新MOR表达/功能，并有选择性潜在的神经胶质介导的炎症 随后在与奖励，认知和 疼痛。该提案中的目标将通过组织实现 伏隔核，腹侧对盖区，前额叶皮层，灰色灰色的测量 背侧海马和杏仁核。 AIM 1将评估MOR状态的纵向变化 放射性结合密度/亲和力以及基因表达（实时PCR）和免疫荧光共同 在TBI/重复吗啡后，在收集7天，1、3和6个月的组织中对神经元和神经胶质的定位。 AIM 2将表征吗啡刺激的经典和炎症途径的差异激活 在TBI和亚少谱吗啡后，长期时间点（3和6个月）的暴露。活动 通过经典途径将通过G蛋白（GTPS）活性和磷酸化ERK的程度评估 免疫荧光。炎症途径信号传导将通过测量神经胶质的程度来评估 免疫荧光，TLR4小胶质染色和流式细胞术和IL-1蛋白的激活 ELISA的数量。假设发现吗啡的潜在炎症反应 遵循TBI将与会议增加与与之相关的不良后果风险 激活这一途径，包括加法脆弱性，认知和痛风障碍，使这些 研究与VA患者护理和康复任务高度相关。这些数据还将提供关键 证据是一项综合研究的基础，以探讨 这些信号结果和药物策略探索了有偏的激动剂。