目的:本研究探讨经Gln处理后对内毒素(LPS)刺激健康人外周血单个核细胞(PBMC)细胞因子表达的影响。方法:取健康志愿者新鲜外周血,用密度梯度离心法提取出单个核细胞,以LPS刺激单个核细胞后,分别使用不同浓度(1、3、6、8、10和15mmol/L)Gin处理2h后,用ELISA法检测细胞上清液中的细胞因子(TNF-α、IL-1B、IL-4、IL-6、IL-8、IL-10)水平,并利用SuperArray基因芯片技术检测细胞因子mRNA。结果:LPS刺激PBMC后,细胞因子大量表达,用Gin处理能显著减少细胞因子的转录和转录后蛋白的表达(P〈0.05),且这一调控作用与Gln剂量呈正相关。结论:Gln能下调LPS刺激后PBMC细胞因子的表达。
Objective : The effect of glutamine on the cytokine expression in peripheral blood mononuclear cells stimulated by lipopolysaccharide was investigated. Methods: We employed the density gradient centrifugation to extract PBMC from fresh peripheral blood of healthy volunteers. The cytokines(TNF-ot, IL-1 α, IL-4, IL-6, IL-8, IL-10)were observed after the PBMC was treated by LPS. Suspension of PBMC was treated with the concentrations of 1, 3, 6, 8, 10, and 15 mmol/L Gin for 4, 12, and 24 h respectively. The expression of the cytokines was detected in the supernatant of PBMC with Enzyme-Linked Im- munosorbnent Assay, and the mRNA of these cytokines was detected in cells with Gene Array Protocol. Results: After LPS stimulation, an excessive expression of the cytokines was observed. These cyto- kines were reduced significantly by glutamine treatment at the transcriptional level and the posttranscrip- tional level (P 〈 O. 05 ). And the attenuation was directly correlated with the dose of Gin. Conclusion: Glutamine can down-regulate the cytokine expression of PBMC stimulated by LPS.