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.

翻译后摘要：虽然很多是了解有关机制的感觉转导从皮肤，相对 对肌肉的这一过程知之甚少。由于反复的 缺血/再灌注（I/R）是影响美国数百万人的主要健康问题， 知识的缺乏使我们无法确定肌肉疼痛或心血管疾病的具体治疗方法。 在缺血的情况下对运动的反应。外周I/R发生在血液疾病中，如镰状细胞 疾病和心血管疾病如外周血管疾病。雄性和雌性展示 在外周血减少的情况下，肌痛和运动加压反射（EPR）的不同特征 灌注。本研究的主要目的是确定肌肉的性别特异性分子机制 可能是从急性缺血性肌痛转变为慢性缺血性肌痛的基础的传入敏化。试点数据 重复性I/R损伤的小鼠模型表明，男性外周致敏受增加的 胶质细胞源性神经营养因子（GDNF）家族受体β 1调控DRG基因表达 (GFR在肌肉伤害感受器中。在女性中，传入敏化、疼痛相关行为和改变的EPR 反复I/R损伤后，IL-1 r1依赖性基因表达增加， 表情我们推测连续性I/R损伤的延长效应是由GDNF介导的 在雄性动物中，IL 1 β诱导了相关的外周致敏作用，在雌性动物中，IL 1 β诱导了致敏作用。目标1将决定是否 肌纤维产生GDNF和DRG上调GFR β 1，调节观察到的肌肉传入变化 男性双重I/R后的反应特性、疼痛相关行为和改变的EPR。我们会用肌肉纤维 特异性GDNF消融或我们新的体内siRNA介导的单个外周神经中基因敲低， 结合我们的离体肌肉传入记录制备或疼痛样行为和EPR的测定。 目标2将利用类似的方法，除了我们将确定巨噬细胞是否产生IL 1 β或DRG IL 1 r1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，目标3将使用离体记录和疼痛相关的行为分析来确定表达是否 富含AU的元件RNA结合蛋白1（AUF 1）或ras家族成员RAN的差异调节了 女性与男性之间连续I/R损伤的性别特异性影响。每一个目标都将是 通过分析DRG和肌肉基因表达来补充。这里概述的研究还将提供一个 肌肉神经生物学研究的新方向，将远远超出目前的增量扩展 报道结果将使我们能够确定与肌肉传入相关的独特的性别依赖机制 I/R损伤后急性至慢性缺血性肌痛发展基础的致敏作用。这可能导致 制定更合适的治疗方法，治疗与缺血相关的肌肉骨骼疼痛或EPR改变/ 靶向适当受体、初级传入亚群或外周靶组织的再灌注。