PV-杀白细胞素和PDTC对THP-1巨噬细胞IL-1β表达的影响

Influence of PVL and PDTC on IL-1β concentration in THP-1

目的探讨NF-κB信号通路在金黄色葡萄球菌PV-杀白细胞毒素相关性肺损伤中的作用。方法实验前用100 nmol·L-1佛波酯孵育THP-1细胞48 h,充分诱导使其分化为巨噬细胞,分为三组,正常对照组、PDTC阻断组和rPVL刺激组,PDTC阻断组实验前1 h加入PDTC孵育,1 h后分别加入PBS和rPVL刺激其诱导分化好的巨噬细胞,采用ELISA检测细胞上清IL-1β的蛋白含量,RT-PCR检测IL-1βmRNA水平的表达,Western-blot检测NF-кB的表达。结果 PV-杀白细胞毒素能促进THP-1巨噬细胞分泌IL-1β,同时rPVL能活化NF-κB蛋白,NF-κB信号通路的特异性阻断剂能够抑制NF-κB蛋白的活化。结论 NF-κB信号通路蛋白激活及细胞因子大量释放可能是PVL相关肺损伤重要的发病机制。

Objective To explore the effects of nuclear transcription factor-κB（NF-κB） signaling pathway proteins and cytokines in Panton-Valentine leukocidin（PVL）-induced acute lung injury.Methods THP-1 cells were cultured in the presence of 100 nmol.L-1 phorbol-12-myristate 3-acetate（PMA） of 48h to induce monocytemacrophage differentiation.After that,THP-1 macrophages were incubated with PVL and PDTC,and then ELISA was performed to test expression of IL-1β in supernatants fluid;RT-PCR was performed to detect expression of IL-1β;NF-κB was detected with Western blotting.Results PVL was able to induce expression of IL-1β in THP-1 macrophages.PVL could also significantly promote the activation of NF-κB.The inhibitor of NF-κB,PDTC,can decrease the secretion of pro-inflammatory in THP-1 macrophages by rPVL.Conclusions NF-κB activation and cytokines release play an important role in PVLrelated lung injury.