谷氨酰胺抑制单个核细胞细胞因子过度表达的研究

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

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。

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.