摘要
目的:探究细胞因子在脂多糖所致脓毒症T淋巴细胞免疫抑制中的作用.方法:以革兰阴性细菌成分脂多糖诱导脓毒症T淋巴细胞免疫抑制,采用流式细胞分析检测对照组和模型组树突状细胞的细胞因子白介素-1α(IL-1α)、IL-6、肿瘤坏死因子-α(TNF-α)、IL-10、转化生长因子-β(TGF-β)、干扰素-α(IFN-α)、IFN-β的表达情况.然后分别阻断树突状细胞的细胞因子进行混合淋巴细胞共培养实验,采用BrdU细胞增殖检测试剂盒检测阻断后T淋巴细胞的免疫增殖抑制的逆转情况.结果:与对照组相比,模型组树突状细胞的细胞因子IL-1α、IL-6、TNF-α、TGF-β、IFN-β阳性百分比显著升高,IL-10、IFN-α阳性百分比无统计学差异.阻断细胞因子后,与模型组相比,αIFN-β+模型组T淋巴细胞增殖反应显著升高.结论:脂多糖所致脓毒症过程中树突状细胞的细胞因子IL-1α、IL-6和TNF-α表达上调可能参与促炎反应过程.细胞因子IFN-β表达上调可能参与淋巴细胞凋亡的免疫抑制过程.阻断IFN-β可改善脂多糖诱导的脓毒症T淋巴细胞增殖抑制现象.
Abstract
Objective:To explore the effect of inflammatory cytokines on the immunosuppression of T lymphocytes in lipopolysaccharide bacterial sepsis.Methods:Gram-negative bacterial component lipopolysaccharide is used to induce dendritic cells to replicate the immuno-suppressive model of bacterial sepsis.Flow cytometry is used to detect the expression of inflammatory cytokines Interleukin-1α(IL-1α),IL-6,tumor necrosis factor-α(TNF-α),IL-10,transforming growth factor-β(TGF-β),interferon-α(IFN-α),IFN-β of dendritic cells in the control group and model group.Then,the BrdU cell proliferation detection kit is used to detect the reversal of immune proliferation inhibition of T lymphocytes in mixed lymphocyte coculture experiments.Results:Compared with the control group,the positive percentages of the inflammatory cytokines IL-1α,IL-6,TNF-α,TGF-β and IFN-β of the dendritic cells in the model group are all significantly increased.But there is no statistical difference of IL-10,IFN-α.Compared with the model group,the T cell proliferation response of the αIFN-β+model group are significantly increased.Conclusion:The up-regulation of inflammatory cytokines IL-1α,IL-6 and TNF-α in dendritic cells may be involved in mediating the pro-inflammatory response process in bacterial sepsis.IFN-β upregulation may be involved in the lymphocyte apoptosis of immune suppression process in sepsis.Importantly IFN-β blocking can significantly reverse the inhibition of T lymphocyte proliferation in lipopolysaccharide-induced immuno-paralysis.
基金项目
河南省基础与前沿技术研究项目(152300410018)
南阳市科技攻关计划项目(2021KJGG118)
NETL
NSTL
万方数据