IL-6、IL-1β和IL-10在外周血的水平可作为早期鉴别革兰阳性与革兰阴性脓毒症的指标

IL-6,IL-1β,and IL-10 levels in peripheral blood as indicators for early identification of gram-positive and gram-negative sepsis

目的通过对脓毒症早期患者外周血单个核细胞(PBMC)转录组测序以及血清液相芯片技术,寻找快速判断革兰阴性(G^(-))和革兰阳性(G^(+))菌的指标.方法选取182例急诊重症监护病房(EICU)救治的脓毒症患者中符合条件的35例进行回顾性分析,按血培养检测结果分为G^(+)组12例和G^(-)组23例,比较两组患者年龄、性别、序贯器官衰竭评分(SOFA)等一般指标,以及血常规、白细胞介素6(IL-6)、C反应蛋白(CRP)、降钙素原(PCT)等化验指标.单次密度梯度离心分离法分离PBMC,提取总RNA进行转录组测序,寻找差异基因.血清液相芯片技术检测两组患者血清粒细胞集落刺激因子(G-CSF)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)、γ干扰素(IFN-γ)、IL-1β、IL-2、IL-4、IL-5、IL-6、IL-7、IL-8、IL-10、IL-12、IL-13、IL-17、单核细胞趋化蛋白1(MCP-1)、肿瘤坏死因子α(TNF-α).将细菌种类作为因变量,以筛选出的细胞因子作为检验变量,绘制受试者工作特征(ROC)曲线,分析所选生物标志物与细菌感染种类的相关性.结果G^(+)组和G^(-)组一般资料及CRP、PCT无明显差异,G^(+)组患者血清IL-6水平较G^(-)组低.转录组测序发现两组患者PBMC约30个免疫相关基因表达有明显差异.与G^(+)组患者相比,G^(-)组患者早期血清IL-6、IL-1β明显升高,IL-10水平下降.ROC曲线分析提示血清IL-6、IL-1β、IL-10水平可以鉴别G^(-)菌、G^(+)菌类型,三者联合诊断对鉴别G^(-)菌、G^(+)菌类型有较高应用价值.结论IL-6、IL-1β和IL-10可以作为脓毒症早期快速判断G^(+)菌或G^(-)菌感染的指标.

Objective To find out indicators for rapid identification of Gram-positive(G^(+))and Gram-negative(G^(-))bacteria through transcriptome sequencing of peripheral blood mononuclear cells(PBMCs)and serum liquid-phase chip technology in early sepsis.Methods 35 eligible cases out of 182 sepsis patients in the emergency intensive care unit(EICU)were selected for retrospective analysis.They were divided into G^(+)group(12 cases)and G^(-)group(23 cases)based on their blood culture results.General characteristics like patients'age,gender,sequential organ failure assessment(SOFA)scores,etc.and other laboratory indexes such as blood routine,IL-6,CRP,procalcitonin(PCT)of these two groups were analyzed.PBMCs were isolated through single density gradient centrifugation.Total RNA was extracted for transcriptome sequencing to find out differential genes.Serum liquid-phase chip technology was performed to detect serum granulocyte colony stimulating factor(G-CSF),granulocyte-macrophage colony stimulating factor(GM-CSF),IFN-γ,IL-1β,IL-2,IL-4,IL-5,IL-6,IL-7,IL-8,IL-10,IL-12,IL-13,IL-17,monocytes chemotactic protein 1(MCP-1),tumor necrosis factor alpha(TNF-α)in two groups.Receiver operating characteristic(ROC)curve was drawn using bacteria types as a dependent variable and selected cytokines as test variables,to analyze the correlation between selected biomarkers and bacteria type.Results No significant difference in general characteristics,CRP,and PCT were found between the G^(+)and G^(-)group.The serum level of IL-6 in G^(+)group was lower than that in the G^(-)group.Transcriptome sequencing results revealed 30 immune-related genes that were differentially expressed in the PBMCs of two groups.Compared to the G^(+)group,the serum levels of IL-6 and IL-1β in G^(-)group significantly increased,while serum IL-10 was reduced.ROC curve analysis indicated that serum IL-6,IL-1β,and IL-10 levels could identify the G^(-)and G^(+)bacteria types.The combined diagnosis using these three indicators is highly applicable in distinguishing G^(-)and G^(+)bacteria.Conclusion IL-6,IL-1β and IL-10 levels can be used as indicators for early identification of sepsis induced by G^(+)or G^(-)bacteria.