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目的探讨硫喷妥钠对CpG DNA诱导人外周血单个核细胞(hPBMC)核因子-κB(NF-κB)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(0.2~1.0 mg/ml),ELISA测定hPBMC培养上清中TNF-α和IL-6浓度,确定硫喷妥钠对CpG DNA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-κB表达。结果不同浓度的硫喷妥钠(0.2~1.0mg/ml)显著抑制CpG DNA诱导hPBMC NF-κB P65表达和TNF-α、IL-6释放(P<0.05或P<0.01)。提前1~4 h给予硫喷妥钠或同时给予硫喷妥钠和CpG DNA(0 h),其拮抗CpG DNA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CpG DNA给予后1 h再加入,也能观察到显著拮抗作用(P<0.05)。结论硫喷妥钠可通过下调CpG DNA诱导hPBMC NF-κB P65表达,从而降低促炎细胞因子TNF-α和IL-6的释放。
Objective To investigate the effects of thiopental on the expression of nuclear factor-κB (NF-κB) and the levels of tumor necrosis factor α (TNF-α), interleukin-6 (IL-6) in human peripheral blood mononuclear cells (hPBMCs) The impact of release. Methods Peripheral blood of healthy volunteers was collected and the hPBMC was separated by density gradient centrifugation using Ficoll-Hypaque. The concentrations of TNF-α and IL-6 in the culture supernatant of hPBMC were determined by ELISA after different concentrations of thiopental (0.2-1.0 mg / ml) were added respectively. The effects of thiopental sodium on secretion of TNF-α by CpG DNA-stimulated cells and IL-6 and its dose-effect relationship and time-effect relationship; Western blotting analysis of hPBMC nuclear NF-κB expression. Results Different concentrations of thiopental (0.2-1.0 mg / ml) significantly inhibited CpG DNA-induced NF-κB P65 expression and TNF-α and IL-6 release in hPBMCs (P <0.05 or P <0.01). Thiopental or thiopental and CpG DNA (0 h) given 1 to 4 h earlier were significantly (P <0.01) antagonistic to CpG DNA-induced cytokine release (P <0.01) Significant antagonism (P <0.05) was also observed when CpG DNA was added 1 h after administration. Conclusion Thiopental inhibits the release of pro-inflammatory cytokines TNF-α and IL-6 by downregulating the expression of NF-κB P65 in hPBMC induced by CpG DNA.