微小RNA-150在脓毒症患者外周血白细胞的表达及其临床意义  

Expression of MicroRNA-150 in Peripheral Blood Leukocytes in Sepsis Patients and Its Clinical Significance

作　　者：曾小莉[1] 张韶岩[2] 张京岚[3] 李凤梅[1] 马雪莲[1] 米玉红[4] 

机构地区：[1]首都医科大学附属北京安贞医院检验科,北京100029 [2]首都医科大学附属北京安贞医院胸外科,北京100029 [3]首都医科大学附属北京安贞医院SICU,北京100029 [4]首都医科大学附属北京安贞医院EICU,北京100029

出　　处：《中国呼吸与危重监护杂志》2011年第4期360-364,共5页Chinese Journal of Respiratory and Critical Care Medicine

基　　金：首都医科大学附属北京安贞医院院长基金资助项目(编号:2010Z09)

摘　　要：目的观察脓毒症患者外周血白细胞中微小RNA-150（miR-150）的表达变化,探讨其与脓毒症免疫细胞因子水平及严重程度的关系。方法采用实时定量PCR（qRT-PCR）检测40例脓毒症患者、20例全身性炎症反应综合征（SIRS）患者和20例正常对照外周血白细胞中miR-150表达量,并评价miRNA检测的稳定性和线性。酶联免疫吸附法测定TNF-α和IL-10浓度,序贯器官衰竭估计（SOFA）评分系统评价脓毒症患者的严重程度。对miR-150与白细胞总数、TNF-α、IL-10和SOFA评分之间进行相关分析。结果外周血标本在4℃保存5 d,白细胞中miR-150表达量无明显变化;最低检出限为6.97 pg/μL RNA,在6.97~6.97×104pg/μL RNA之间有良好的线性关系（r=0.999）。脓毒症组miR-150表达量较正常对照组显著降低（P〈0.01）,白细胞总数、IL-10水平和IL-10/TNF-α比值显著升高（P〈0.05）;而脓毒症组与SIRS组间差异均无统计学意义（P〉0.05）。血清TNF-α水平在三组间差异均无统计学意义（P〉0.05）。脓毒症患者中,死亡组miR-150表达量较存活组显著降低（P〈0.05）,而IL-10水平和IL-10/TNF-α比值显著升高（P〈0.01）,TNF-α水平差异无统计学意义（P〉0.05）。脓毒症患者miR-150表达量与SOFA评分、血清TNF-α和IL-10呈显著负相关（r值分别为-0.619、-0.457和-0.431,P〈0.05）;与WBC无显著相关性（P〉0.05）。血清TNF-α、IL-10和IL-10/TNF-α比值与SOFA评分无显著相关性（P〉0.05）。结论脓毒症患者外周血miR-150表达量显著降低,其水平不但能反映机体炎症反应状态,而且在一定程度上能作为判断疾病严重程度及预后的指标。Objective To investigate the changes of microRNA-150（miR-150） in peripheral blood leukocytes in sepsis patients,and their relationship with expression of immune cytokines and sepsis severity.Methods The level of mature miR-150 was quantified by real-time reverse transcriptase-polymerase chain reaction（RT-PCR） and normalized to that of control miRNA,U6,in peripheral blood leukocytes of 40 patients with sepsis,20 patients with systemic inflammatory response syndrome（SIRS）,and 20 normal individuals.Serum concentrations of tumor necrosis factor alpha（TNF-α） and interleukin-10（IL-10） were measured by enzyme-linked immunoabsorbent assay in all subjects.The sequential organ failure assessment（SOFA） score system was used to evaluate the severity of sepsis.The relationships between miR-150 and the white blood cell count（WBC）,TNF-α,IL-10 and SOFA score of the sepsis patients were analyzed.Results MiR-150 was stable for at least 5 days when specimen stored at 4 ℃ and the determination of miR-150 had a broad linear detecting range（6.97-6.97×104 pg/μL RNA,the lowest detecting limit：6.97 pg/μL RNA,r=0.999）.MiR-150 expression in the peripheral blood leukocytes in the sepsis group was significantly lower than that in the healthy control group（P0.01）,while WBC,IL-10 and IL-10/TNF-α ratio were significantly higher（P0.05）.There was no significant difference in levels of miR-150,IL-10,IL-10/TNF-α ratio,and WBC between the sepsis group and the SIRS group（P0.05）.There was no significant difference in serum concentrations of TNF-α among three groups（P0.05）.MiR-150 expression in non-survivor sepsis patients was significantly lower than that in survivor sepsis patients（P0.05）,while serum IL-10 and IL-10/TNF-α ratio were significantly higher（P0.01）,but there was no significant difference in serum TNF-α between the non-survivor group and the survivor group（P0.05）.There was significantly negative correlation between miR-150 and SOFA score,TNF-α and IL-10（r=-0.619,-

关 键 词：脓毒症 微小RNA-150 肿瘤坏死因子Α 白细胞介素10 序贯器官衰竭估计评分 

分 类 号：R459.7[医药卫生—急诊医学]