Department of Evolutionary Immunobiology, Institute of Zoology, Jagiellonian University, Cracow, Poland.
Arch Immunol Ther Exp (Warsz). 2012 Oct;60(5):373-82. doi: 10.1007/s00005-012-0186-x. Epub 2012 Aug 23.
Zymosan-induced peritonitis represents a well-described model of acute inflammation. The binding of zymosan with its specific Toll-like receptors (TLR2 and TLR6) on leukocytes initiates activation and phosphorylation of nuclear factor (NF)-κB, which leads to accumulation of NF-κB p65 subunits in the nucleus and subsequently up-regulation of the proinflammatory cytokine genes expression. Intraperitoneal co-administration of zymosan and morphine significantly inhibits peritonitis in several strains of mice by decreasing the influx of exudatory cells; however, mechanisms of this action still remain unclear. We aimed to verify the effects of morphine on NF-κB and TLRs expression at messenger RNA and protein levels during the early stages of zymosan-induced peritonitis. Peritonitis was induced by a single injection of zymosan A or zymosan supplemented with morphine in Swiss mice. At selected time points, after stimulation, peritoneal leukocytes were harvested. The TLRs and NF-κB expression was assessed by real-time PCR and flow cytometry. In comparison with the mice injected with zymosan only, morphine co-injection significantly decreased the expression of phospho-NF-κB and TLR2 in all investigated immunocompetent cells as well as up-regulated the levels of nitric oxide (NO) in peritoneal fluid. Moreover, supplementation of zymosan with morphine altered the TLR, NF-κB and some proinflammatory cytokines (keratinocyte-derived chemokine, tumor necrosis factor-α) gene expression during ongoing inflammation. We may postulate that after morphine stimulation peritoneal leukocytes recognize less effectively zymosan antigens because of impaired TLRs expression. The lower TLR expression attenuates TLR-mediated signal transduction, which prevents NF-κB activation. Additionally, during zymosan-induced peritonitis, morphine may modulate the NF-κB expression, at least partially, by an up-regulated release of NO, as suggested by others.
酵母聚糖诱导的腹膜炎是一种典型的急性炎症模型。酵母聚糖与白细胞上的特定 Toll 样受体（TLR2 和 TLR6）结合，引发核因子（NF）-κB 的激活和磷酸化，导致 NF-κB p65 亚基在核内积累，并随后上调促炎细胞因子基因的表达。酵母聚糖和吗啡腹腔内共同给药可显著抑制几种品系小鼠的腹膜炎，减少渗出细胞的流入；然而，其作用机制仍不清楚。我们旨在验证吗啡在酵母聚糖诱导的腹膜炎早期对 NF-κB 和 TLRs 表达的影响，检测 mRNA 和蛋白水平。通过单次注射酵母聚糖 A 或添加吗啡的酵母聚糖诱导腹膜炎，在选定的时间点，刺激后收集腹膜白细胞。通过实时 PCR 和流式细胞术评估 TLRs 和 NF-κB 的表达。与单独注射酵母聚糖的小鼠相比，吗啡共注射显著降低了所有研究免疫活性细胞中磷酸化 NF-κB 和 TLR2 的表达，并上调了腹腔液中一氧化氮（NO）的水平。此外，吗啡对酵母聚糖的补充改变了 TLR、NF-κB 和一些促炎细胞因子（角质形成细胞衍生趋化因子、肿瘤坏死因子-α）在持续炎症过程中的基因表达。我们可以假设，在吗啡刺激后，由于 TLR 表达受损，腹膜白细胞对酵母聚糖抗原的识别效果降低。较低的 TLR 表达减弱了 TLR 介导的信号转导，从而阻止了 NF-κB 的激活。此外，在酵母聚糖诱导的腹膜炎中，吗啡可能通过上调一氧化氮的释放来调节 NF-κB 的表达，正如其他人所建议的那样。
