摘要
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.
出处
《临床麻醉学杂志》
CAS
CSCD
2008年第2期103-105,共3页
Journal of Clinical Anesthesiology