谷氨酰胺抑制单个核细胞细胞因子过度表达的研究

目的:探讨谷氨酰胺(Gln)抑制人外周血单个核细胞(PBMC)细胞因子过度表达的分子机制。方法:用Ficoll密度梯度离心法提取健康志愿者新鲜外周血单个核细胞,在培养液中加入Gln、P38MAPK通路阻滞剂(SB203580)和HSP70阻断剂(Quercetin),用内毒素刺激4 h后,留取细胞和上清液,ELISA法测定内毒素刺激下单个核细胞肿瘤坏死因子(TNF-α)和IL-10的表达,Western blot方法检测细胞内磷酸化P38的表达情况。结果:Gln能明显增加HSP70的表达,同时TNF-α和IL-10表达受抑;使用Quercetin和SB203580后,磷酸化P38减少的同时TNF-α和IL-10也减少了。结论:Gln下调LPS刺激下PBMC细胞因子的过度表达依赖于P38MAPK信号蛋白的抑制。

Mechanism of glutaminl downregulates the cytokine expression in lipopolysaccharide-stimulated human peripheral blood mononuclear cells

Objective: To investigate the mechanism that glutamine downregulates the cytokine expression in lipopolysaccharide-stimulated human peripheral blood mononuclear cells.Methods:PMBC were extracted from healthy volunteer by density gradient centrifugation,and stimulated by lipopolysaccharide.The release of TNF-α and IL-10 was analyzed via enzyme-linked immunosorbent assay and p-P38 via western blot. Results:Gln led to an increase in HSP70 expression,and decreased TNF-α、IL-10 release at 4 h after LPS stimulation.The expressions level of TNF-α、IL-10 and P-P38 were decreased after using SB203580.Conclusion:These results indicate that the effect of Gln could attenuate cytokine release in PBMC and might may be related to the inhibition of P38 MAPK.