Vilekar Prachi, Rao Geeta, Awasthi Shanjana, Awasthi Vibhudutta
Department of Pharmaceutical Sciences, College of Pharmacy, University of Oklahoma Health Sciences Center, 1110 North Stonewall Avenue, Oklahoma City, OK 73117 USA.
J Inflamm (Lond). 2015 Sep 23;12:55. doi: 10.1186/s12950-015-0096-x. eCollection 2015.
Unresolved and prolonged inflammation is a pathological basis of many disorders such as cancer and multiple organ failure in shock. Interleukin-1 receptor (IL-1R) superfamily consists of IL-1R1 and pathogen pattern recognition receptor toll-like receptor-4 (TLR4) which, upon ligand binding, initiate pro-inflammatory signaling. The study objective was to investigate the effect of a diphenyldifluoroketone EF24 on the expression of IL-1R1 and TLR4 in lipopolysaccharide (LPS)-stimulated dendritic cells (DCs).
Immortalized murine bone marrow-derived JAWS II dendritic cells (DC) were challenged with LPS (100 ng/ml) for 4 h. The LPS-stimulated DCs were treated with 10 μM of EF24 for 1 h. The expression levels of IL-1R1 and TLR4 were monitored by RT-PCR, immunoblotting, and confocal microscopy. The effect of EF24 on the viability and cell cycle of DCs was examined by lactate dehydrogenase assay and flow cytometry, respectively.
EF24 treatment suppressed the LPS-induced TLR4 and IL-1R1 expression in DCs. However, the expression levels of IL-1RA and IL-1R2 were not influenced by either LPS or EF24 treatments. These effects of EF24 were associated with a decrease in LPS-induced expression of phospho-NF-kB p65, indicative of its role in the transcriptional control of IL-1R superfamily members. We did not find any significant effect of EF24 on the proliferation or cell cycle of DCs.
The results suggest that EF24 influences IL-1R superfamily signaling pathway in ways that could have salutary effects in inflammation. The pluripotent anti-inflammatory actions of EF24 warrant further investigation of EF24 in inflammatory conditions of systemic nature.
炎症未消退且持续时间延长是许多疾病（如癌症和休克中的多器官功能衰竭）的病理基础。白细胞介素-1受体（IL-1R）超家族由IL-1R1和病原体模式识别受体Toll样受体4（TLR4）组成，它们在与配体结合后启动促炎信号传导。本研究的目的是探讨二苯基二氟甲酮EF24对脂多糖（LPS）刺激的树突状细胞（DCs）中IL-1R1和TLR4表达的影响。
用LPS（100 ng/ml）刺激永生化小鼠骨髓来源的JAWS II树突状细胞（DC）4小时。用10 μM的EF24处理LPS刺激的DCs 1小时。通过逆转录聚合酶链反应（RT-PCR）、免疫印迹和共聚焦显微镜监测IL-1R1和TLR4的表达水平。分别通过乳酸脱氢酶测定和流式细胞术检测EF24对DCs活力和细胞周期的影响。
EF24处理抑制了DCs中LPS诱导的TLR4和IL-1R1表达。然而，IL-1RA和IL-1R2的表达水平不受LPS或EF24处理的影响。EF24的这些作用与LPS诱导的磷酸化核因子-κB p65表达降低有关，表明其在IL-1R超家族成员转录控制中的作用。我们未发现EF24对DCs的增殖或细胞周期有任何显著影响。
结果表明，EF24以可能对炎症有益的方式影响IL-1R超家族信号通路。EF24的多能抗炎作用值得在全身性炎症条件下对其进行进一步研究。
