硫喷妥钠对CpG DNA诱导人外周血单个核细胞NF-κB表达和TNF-α、IL-6释放的影响

Effects of thiopental on NF-κB expression and TNF-α,IL-6 release from human peripheral blood mononuclear cells induced by CpG DNA

目的探讨硫喷妥钠对CpGDNA诱导人外周血单个核细胞（hPBMc）核因子-kB（NF-kB）表达及肿瘤坏死因子α（TNF-α）、白细胞介素-6（IL-6）释放的影响。方法采集健康志愿者外周血，使用淋巴细胞分离液（FiColl—Hypaque）密度梯度离心分离hPBMC。分别加入不同浓度的硫喷妥钠后（0．2～1．0mg／ml），ELISA测定hPBMC培养上清中TNF-α和IL-6浓度，确定硫喷妥钠对CpGDNA刺激细胞分泌TNF-α及IL-6的抑制作用及其剂量-效应关系和时间-效应关系；免疫蛋白印迹法（Western blotting）分析hPBMC胞核NF—KB表达。结果不同浓度的硫喷妥钠（0．2～1.0mg／ml）显著抑制CF）GDNA诱导hPBMC NF—kB P65表达和TNF-α、IL-6释放（P〈0．05或P〈0．01）。提前1～4h给予硫喷妥钠或同时给予硫喷妥钠和CpGDNA（0h），其拮抗CpGDNA诱导细胞因子释放的作用非常显著（P〈0．01），且硫喷妥钠在刺激物CpGDNA给予后1h再加入，也能观察到显著拈抗作用（P＜0．05）。结论硫喷妥钠可通过下调CpGDNA诱导hPBMCNF—kB P65表达，从而降低促炎细胞因子TNF-α和IL-6的释放。

Objective To explore the effects of thiopental on nuclear factor-kB （NF-kB） expression,and release tumor necrosis faetor-α （TNF-α） and interleukin-6 （IL-6） from human peripheral blood mononuelear cells （hPBMCs） induced by CpG DNA. Methods hPBMCs from healthy people were separated by Fieoll-Hypaque density gradient eentrifugation. TNF-α and IL-6 were detected by ELISA after adding different concentrations of thiopental （0.2-1.0 mg/ml） for determining the dose- and time-dependent manner of its inhibitory effect. NF-kB P65 protein in hPBMC was analyzed by Western blotting. Results Thiopental with different concentrations （0.2- 1.0 g/L） inhibited markedly TNF-α and IL-6 release and NF-kB P65 protein expression in hPBMCs stimulated by CpG DNA（P〈0.05 or P〈0.01）. Addition of thiopental 1-4 h prior to CpG DNA （P〈 0.01） or coincident with CpG DNA stimulation （0 h） attenuated significantly the release of TNF-α and IL-6. The release of TNF-α and IL-6 was also inhibited by thiopental 1 h after addition of stimulator CpG DNA. Conclusion Thiopental can attenuate TNF-α and IL-6 release in hPBMCs through down regulating the expression of nuclear transcriptional factor NF-kB P65.