Mechanisms of muscle afferent sensitization after ischemia

缺血后肌肉传入敏化机制

• 批准号: 10271290
• 负责人: Michael P Jankowski
• 金额: $ 48.88万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10271290
• 关键词: Ablation; Acute; Affect; Afferent Neurons; Automobile Driving; Behavior; Behavioral; Cardiovascular Diseases; Cardiovascular system; Cell Line; Cell Nucleus; Cells; Chronic; Complement; Data; Development; Disease; Electrophysiology (science); Elements; Exercise; Family; Family member; Female; Formulation; Gene Expression; Genes; Goals; Growth Factor; Health; Hematological Disease; Immune; Individual; Injury; Interleukin-1; Interleukin-1 Receptors; Ischemia; Knock-out; Knowledge; Lead; Mediating; Molecular; Mus; Muscle; Muscle Fibers; Musculoskeletal Pain; Myalgia; Nerve; Neurobiology; Nociceptors; Outcome; Pain; Pain Measurement; Pain Threshold; Pathway interactions; Perfusion; Peripheral; Peripheral Nerves; Peripheral Vascular Diseases; Persistent pain; Play; Population; Preparation; Prevalence; Process; Property; RNA Sequences; RNA-Binding Proteins; Reflex action; Reperfusion Injury; Reperfusion Therapy; Reporting; Research; Role; Sex Differences; Sickle Cell Anemia; Signal Transduction; Skeletal Muscle; Skin; Small Interfering RNA; Source; Spinal Cord; Spinal Ganglia; Testing; Tissues; Transgenic Organisms; United States; Up-Regulation; base; behavioral response; cytokine; experience; experimental study; glial cell-line derived neurotrophic factor; in vivo; innovation; insight; knock-down; macrophage; male; member; mouse model; neurotrophic factor; novel; novel therapeutics; prevent; ras Oncogene; receptor; response; sensory mechanism; sex; transcription factor

Abstract: While great deal is understood about mechanisms of sensory transduction from the skin, relatively little is known about this process from the muscles. As peripheral injuries resulting from repetitive ischemic/reperfusion (I/R) are a major health issue that affects millions of people in the United States, this gap in knowledge precludes us from identifying specific therapies for muscle pain or altered cardiovascular responses to exercise in the context of ischemia. Peripheral I/R occurs in blood disorders such as sickle cell disease, and in cardiovascular disorders such as peripheral vascular disease. Males and females display different features of myalgia and exercise pressor reflexes (EPRs) under conditions of reduced peripheral perfusion. The major goal of this study is to determine the sex specific molecular mechanisms of muscle afferent sensitization that may underlie the transition from acute to chronic ischemic myalgia. Pilot data in murine models of repetitive I/R injury suggest that peripheral sensitization in males is regulated by increased DRG gene expression that is modulated by glial cell line-derived neurotrophic factor (GDNF) family receptor 1 (GFR1) in muscle nociceptors. In females, afferent sensitization, pain-related behaviors and altered EPRs after repeated I/R injury may be regulated by increased interleukin 1 receptor type 1 (IL1r1) dependent gene expression. We hypothesize that the prolonged effects of successive I/R injuries are mediated by GDNF related peripheral sensitization in males and IL1 induced sensitization in females. Aim 1 will determine if myofiber produced GDNF and DRG upregulation of GFR1, regulate the observed changes in muscle afferent response properties, pain-related behaviors and altered EPRs after dual I/R in males. We will use muscle fiber specific GDNF ablation or our novel in vivo siRNA-mediated knockdown of genes in single peripheral nerves in conjunction with our ex vivo muscle afferent recording preparations or assays of pain-like behaviors and EPRs. Aim 2 will utilize a similar approach except we will determine if macrophage produced IL1 or DRG upregulation of IL1r1, regulate the novel changes in female muscle afferents or behaviors after successive I/R. Finally, Aim 3 will use ex vivo recording and pain-related behavioral analyses to determine if expression differences in AU-rich element RNA-binding protein 1 (AUF1) or the ras family member, RAN, modulates the sex specific effects of successive I/R injuries between females versus males. Each of these aims will be complemented by analysis of DRG and muscle gene expression. The studies outlined here will also provide a novel direction in muscle neurobiology research that will go well beyond the incremental expansion of current reports. Results will enable us to identify unique sex dependent mechanisms associated with muscle afferent sensitization that underlie acute to chronic ischemic myalgia development after I/R injury. This may lead to the formulation of more appropriate treatments for musculoskeletal pain or altered EPRs associated with ischemia/ reperfusion that target the proper receptor(s), primary afferent subpopulation(s) or peripheral target tissues.

摘要：虽然人们对皮肤感觉传导机制有很多了解，但相对而言 人们对肌肉的这一过程知之甚少。由于重复性的周围损伤 缺血/再灌注 (I/R) 是影响美国数百万人的主要健康问题，这种差距 知识的匮乏使我们无法确定针对肌肉疼痛或改变心血管的具体疗法 对缺血情况下运动的反应。外周 I/R 发生在镰状细胞病等血液疾病中 疾病，以及心血管疾病，例如周围血管疾病。男性和女性展示 外周血减少条件下肌痛和运动加压反射 (EPR) 的不同特征 灌注。本研究的主要目标是确定肌肉的性别特异性分子机制 传入敏化可能是急性缺血性肌痛向慢性缺血性肌痛转变的基础。试点数据 重复性 I/R 损伤的小鼠模型表明，雄性的外周敏化受到增加的调节 DRG 基因表达受神经胶质细胞源性神经营养因子 (GDNF) 家族受体 α1 调节 (GFRα1) 存在于肌肉伤害感受器中。在女性中，传入敏化、疼痛相关行为和 EPR 改变 反复 I/R 损伤后可能受到白细胞介素 1 1 型受体 (IL1r1) 依赖性基因增加的调节 表达。我们假设连续 I/R 损伤的长期影响是由 GDNF 介导的 男性中相关的外周敏化和女性中 IL1 诱导的敏化。目标 1 将确定是否 肌纤维产生 GDNF 和 DRG 上调 GFRα1，调节观察到的肌肉传入变化 男性双重 I/R 后的反应特性、疼痛相关行为和 EPR 改变。我们将使用肌肉纤维 特异性 GDNF 消融或我们新颖的体内 siRNA 介导的单个外周神经基因敲低 与我们的离体肌肉传入记录准备或疼痛样行为和 EPR 分析相结合。 目标 2 将采用类似的方法，但我们将确定巨噬细胞是否产生 IL1 或 DRG IL1r1 的上调，调节连续 I/R 后女性肌肉传入或行为的新变化。 最后，Aim 3 将使用离体记录和疼痛相关行为分析来确定表达是否 富含 AU 元件的 RNA 结合蛋白 1 (AUF1) 或 ras 家族成员 RAN 的差异调节 女性与男性之间连续 I/R 损伤的性别特异性影响。这些目标中的每一个都将 辅以 DRG 和肌肉基因表达分析。这里概述的研究还将提供 肌肉神经生物学研究的新方向将远远超出当前的增量扩展 报告。结果将使我们能够识别与肌肉传入相关的独特的性别依赖性机制 I/R 损伤后急性至慢性缺血性肌痛发展的致敏作用。这可能会导致 制定更合适的治疗肌肉骨骼疼痛或改变与缺血相关的 EPR 的方法/ 针对适当受体、初级传入亚群或外周靶组织的再灌注。