Interleukin-4 Induces the Release of Opioid Peptides from M1 Macrophages in Pathological Pain
Interleukin-4 Induces the Release of Opioid Peptides from M1 Macrophages in Pathological Pain
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DOI：
10.1523/jneurosci.3040-20.2021
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发表时间：
2021-03-31
影响因子：
5.3
通讯作者：
Machelska, Halina
中科院分区：
文献类型：
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作者：
Labuz, Dominika;Celik, Melih O.;Machelska, Halina
白细胞介素-4（IL-4）是一种抗炎细胞因子，可缓解疼痛。IL-4介导的作用被认为主要是由于抑制促炎介质（例如，IL-1 α 1、肿瘤坏死因子、前列腺素E2）。在这里，我们发现，IL-4注射在损伤的神经减轻疼痛，从小鼠浸润的巨噬细胞释放阿片肽。阿片类物质由IL-4以细胞内Ca 21依赖性方式分泌，并激活局部外周阿片受体。这些作用代表了IL-4作用的一种新模式，因为其释放特性迄今尚未报道。重要的是，我们的研究结果表明，IL-4？白细胞介素-4（IL-4）是一种抗炎细胞因子，在炎症和神经系统疾病中具有保护作用，并能减轻疼痛。传统上，IL-4通过阻断促炎细胞因子的产生来减轻疼痛。在这里，我们发现，IL-4诱导急性抗伤害感受的IL-4受体（IL-4 Ra）依赖性释放阿片肽从M1巨噬细胞在受损的神经。作为病理性疼痛的模型，我们使用雄性小鼠坐骨神经的慢性压迫性损伤（CCI）。单次应用IL-4在损伤的神经（CCI后14天）衰减机械敏感性评价冯弗雷丝，这是逆转共注射抗体IL-4 Ra，抗体阿片肽，如甲硫氨酸脑啡肽（ENK），b-内啡肽和强啡肽A1？17，以及d-阿片样物质、m-阿片样物质和j-阿片样物质受体的选择性拮抗剂。分别通过流式细胞术和qRT-PCR评估，受损神经主要被促炎性M1巨噬细胞浸润，IL-4未改变其数量或表型。通过免疫磁性分离（IMS）从受损神经中分离的巨噬细胞用IL-4剂量依赖性地刺激分泌所有三种通过免疫测定测量的阿片肽。IL-4 Ra抗体、细胞内Ca ~（21）螯合剂、蛋白激酶A（PKA）抑制剂、磷酸肌醇3-激酶（PI 3 K）抑制剂和ryanodine受体抑制剂可抑制IL-4诱导的ENK释放。总之，我们确定了一个新的阿片类药物的机制，涉及PKA介导的，PI 3 K介导的，ryanodine受体介导的，和细胞内Ca 21介导的释放阿片肽的M1巨噬细胞的IL-4诱导的抗伤害性，激活外周阿片受体在受伤的组织。
Interleukin-4 (IL-4) is an anti-inflammatory cytokine, which can ameliorate pain. The IL-4-mediated effects are considered to mostly result from the inhibition of the production of proinflammatory mediators (e.g., IL-1Ali , tumor necrosis factor, prostaglandin E2). Here, we found that IL-4 injected at the injured nerves attenuates pain by releasing opioid peptides from the infiltrating macrophages in mice. The opioids were secreted by IL-4 in the intracellular Ca21-dependent manner and activated local peripheral opioid receptors. These actions represent a novel mode of IL-4 action, since its releasing properties have not been so far reported. Importantly, our findings suggest that the IL-4?opioid system should be targeted in the peripheral damaged tissue, since this can be devoid of central and systemic side effects.Interleukin-4 (IL-4) is an anti-inflammatory cytokine, which can be protective in inflammatory and neurologic disorders, and can alleviate pain. Classically, IL-4 diminishes pain by blocking the production of proinflammatory cytokines. Here, we uncovered that IL-4 induces acute antinociception by IL-4 receptor a (IL-4Ra)-dependent release of opioid peptides from M1 macrophages at injured nerves. As a model of pathologic pain, we used a chronic constriction injury (CCI) of the sciatic nerve in male mice. A single application of IL-4 at the injured nerves (14 d following CCI) attenuated mechanical hypersensitivity evaluated by von Frey filaments, which was reversed by co-injected antibody to IL-4Ra, antibodies to opioid peptides such as Met-enkephalin (ENK), b-endorphin and dynorphin A 1?17, and selective antagonists of d-opioid, m-opioid, and j-opioid receptors. Injured nerves were predominately infiltrated by proinflammatory M1 macrophages and IL-4 did not change their numbers or the phenotype, assessed by flow cytometry and qRT-PCR, respectively. Macrophages isolated from damaged nerves by immunomagnetic separation (IMS) and stimulated with IL-4 dose dependently secreted all three opioid peptides measured by immunoassays. The IL-4-induced release of ENK was diminished by IL-4Ra antibody, intracellular Ca21 chelator, and inhibitors of protein kinase A (PKA), phosphoinositide 3-kinase (PI3K), and ryanodine receptors. Together, we identified a new opioid mechanism underlying the IL-4-induced antinociception that involves PKA-mediated, PI3K-mediated, ryanodine receptor-mediated, and intracellular Ca21-mediated release from M1 macrophages of opioid peptides, which activate peripheral opioid receptors in injured tissue.