Newville Jessie, Maxwell Jessie R, Kitase Yuma, Robinson Shenandoah, Jantzie Lauren L
Department of Neurosciences, University of New Mexico School of Medicine, Albuquerque, NM, United States.
Departments of Pediatrics, University of New Mexico School of Medicine, Albuquerque, NM, United States.
Front Pediatr. 2020 Jun 26;8:272. doi: 10.3389/fped.2020.00272. eCollection 2020.
The increased incidence of opioid use during pregnancy warrants investigation to reveal the impact of opioid exposure on the developing fetus. Exposure during critical periods of development could have enduring consequences for affected individuals. Particularly, evidence is mounting that developmental injury can result in immune priming, whereby subsequent immune activation elicits an exaggerated immune response. This maladaptive hypersensitivity to immune challenge perpetuates dysregulated inflammatory signaling and poor health outcomes. Utilizing an established preclinical rat model of perinatal methadone exposure, we sought to investigate the consequences of developmental opioid exposure on activation of peripheral blood mononuclear cells (PBMCs). We hypothesize that PBMCs from methadone-exposed rats would exhibit abnormal chemokine and cytokine expression at baseline, with exaggerated chemokine and cytokine production following immune stimulation compared to saline-exposed controls. On postnatal day (P) 7, pup PMBCs were isolated and cultured, pooling three pups per . Following 3 and 24 h, the supernatant from cultured PMBCs was collected and assessed for inflammatory cytokine and chemokine expression at baseline or lipopolysaccharide (LPS) stimulation using multiplex electrochemiluminescence. Following 3 and 24 h, baseline production of proinflammatory chemokine and cytokine levels were significantly increased in methadone PBMCs ( < 0.0001). Stimulation with LPS for 3 h resulted in increased tumor necrosis factor (TNF-α) and C-X-C motif chemokine ligand 1 (CXCL1) expression by 3.5-fold in PBMCs from methadone-exposed PBMCs compared to PBMCs from saline-exposed controls ( < 0.0001). Peripheral blood mononuclear cell hyperreactivity was still apparent at 24 h of LPS stimulation, evidenced by significantly increased TNF-α, CXCL1, interleukin 6 (IL-6), and IL-10 production by methadone PMBCs compared to saline control PBMCs ( < 0.0001). Together, we provide evidence of increased production of proinflammatory molecules from methadone PBMCs at baseline, in addition to sustained hyperreactivity relative to saline-exposed controls. Exaggerated peripheral immune responses exacerbate inflammatory signaling, with subsequent consequences on many organ systems throughout the body, such as the developing nervous system. Enhanced understanding of these inflammatory mechanisms will allow for appropriate therapeutic development for infants who were exposed to opioids during development. Furthermore, these data highlight the utility of this PBMC assay technique for future biomarker development to guide specific treatment for patients exposed to opioids during gestation.
孕期阿片类药物使用的增加促使人们展开调查，以揭示阿片类药物暴露对发育中胎儿的影响。在发育的关键时期暴露可能会对受影响个体产生持久后果。特别是，越来越多的证据表明，发育损伤会导致免疫致敏，即随后的免疫激活会引发过度的免疫反应。这种对免疫挑战的适应不良的超敏反应会使炎症信号失调，并导致不良健康后果持续存在。我们利用一种已建立的围产期美沙酮暴露的临床前大鼠模型，试图研究发育性阿片类药物暴露对外周血单核细胞（PBMC）激活的影响。我们假设，与生理盐水暴露的对照组相比，来自美沙酮暴露大鼠的PBMC在基线时会表现出趋化因子和细胞因子表达异常，在免疫刺激后趋化因子和细胞因子产生会过度增加。在出生后第7天（P7），分离并培养幼崽的PBMC，每组合并三只幼崽。在3小时和24小时后，收集培养的PBMC的上清液，并使用多重电化学发光法评估基线或脂多糖（LPS）刺激下的炎症细胞因子和趋化因子表达。在3小时和24小时后，美沙酮PBMC中促炎趋化因子和细胞因子水平的基线产生显著增加（P<0.0001）。与生理盐水暴露的对照组的PBMC相比，用LPS刺激3小时后，美沙酮暴露的PBMC中肿瘤坏死因子（TNF-α）和C-X-C基序趋化因子配体1（CXCL1）的表达增加了3.5倍（P<0.0001）。在LPS刺激24小时时，外周血单核细胞的高反应性仍然明显，与生理盐水对照的PBMC相比，美沙酮PBMC中TNF-α、CXCL1、白细胞介素6（IL-6）和IL-10的产生显著增加（P<0.0001）。总之，我们提供了证据表明，美沙酮PBMC在基线时促炎分子的产生增加，并且相对于生理盐水暴露的对照组持续存在高反应性。过度的外周免疫反应会加剧炎症信号，随后会对全身许多器官系统产生影响，如发育中的神经系统。对这些炎症机制的进一步了解将有助于为发育期间暴露于阿片类药物的婴儿开发适当的治疗方法。此外，这些数据突出了这种PBMC检测技术在未来生物标志物开发中的实用性，以指导对孕期暴露于阿片类药物的患者进行特定治疗。
