Wu Yuhui, Li Chengrong, He Yanxia, Li Qiu, Wang Guobing, Wen Pengqiang, Yang Weiguo, Yang Yanlan
Pediatric Intensive Care Unit, Shenzhen Children's Hospital of Chongqing Medical University, Shenzhen 518026, China.
Institute of Pediatrics, Shenzhen Children's Hospital of Chongqing Medical University, Shenzhen 518026, China. Email:
Zhonghua Er Ke Za Zhi. 2014 Jan;52(1):28-33.
Sepsis is the major cause of death in pediatric intensive care unit (PICU). The clinical manifestations of early sepsis is very similar to systemic inflammatory response syndrome (SIRS) caused by non-infectious reason. This study aimed to investigate the expression of miRNA and inflammatory cytokines in plasma in pediatric sepsis patients and its clinical significance.
Forty children with sepsis seen in Shenzhen children's hospital PICU from April 2012 to March 2013 were enrolled in this study, the median age was 0.75 (0.52, 1.90) years; 27 were males and 13 females, of whom 16 had severe sepsis. We selected 20 postsurgical patients with SIRS and 15 healthy children as a control group. The expression levels of plasma miR-21, miR-125b, miR-132, miR-146a, miR-155 and miR-223 were detected by real-time quantitative PCR (qRT-PCR). The predictive value of miRNA, PCT and CRP for sepsis were evaluated by Receiver operating characteristic curve (ROC). TNF-α and IL-10 levels in plasma detected by Cytometric Beads Array (CBA). Quantitative data of normal distribution was compared with ANOVA among the three groups and LSD-t test between two groups. To non-normal distribution of data, multiple comparisons among three groups were conducted by Kruskal-Wallis H test and differences between two groups were assessed by Mann-Whitney U test for post hoc analysis.
There were no significant differences between the age and gender of each group. Expression of miR-21, miR-125b, miR-132 and miR-155 in plasma had no significant difference in each group (all P > 0.05). MiR-146a and miR-223 levels in sepsis were upregulated compared with SIRS group and control group [(5.7 ± 3.5)×10(-5) vs. (2.4 ± 1.6)×10(-5) and (2.6 ± 1.2)×10(-5), (12.5 ± 7.7)×10(-4) vs. (8.3 ± 3.4)×10(-4) and (5.3 ± 2.2)×10(-4), all P < 0.01], expression levels of miR-223 in SIRS increased as compared to control group (P < 0.01). MiR-146a levels in severe sepsis were higher than those of the general sepsis [ (7.1 ± 3.3)×10(-5) vs. (4.6 ± 2.6)×10(-5), P < 0.01]. CRP and PCT levels are all higher in sepsis and SIRS groups than control group (all P < 0.01). The area under ROC curve (AUC) of miR-146a, miR-223, PCT and CRP to predict sepsis were 0.815 (95%CI: 0.708-0.922), 0.678(95%CI: 0.537-0.818), 0.706 (95%CI: 0.571-0.842) and 0.588 (95%CI: 0.427-0.748). Expression levels of IL-10 and IL-10/TNF-α in sepsis were upregulated compared with and SIRS group and the control group (all P < 0.01). There was a positive correlation between miR-146a, miR-223 and IL-10 and IL-10/TNF-α (r = 0.545, 0.305, 0.562, 0.373, all P < 0.01).
The expression levels of miR-146a and miR-223 in plasma in pediatric patients with sepsis was significantly upregulated, and had a positive correlation with IL-10 and IL-10/TNF-α, which may be used as early diagnostic markers and can reflect the severity of condition to a certain degree.
脓毒症是儿科重症监护病房（PICU）死亡的主要原因。早期脓毒症的临床表现与非感染性原因引起的全身炎症反应综合征（SIRS）非常相似。本研究旨在探讨小儿脓毒症患者血浆中miRNA和炎症细胞因子的表达及其临床意义。
选取2012年4月至2013年3月在深圳市儿童医院PICU就诊的40例脓毒症患儿，中位年龄为0.75（0.52，1.90）岁；男27例，女13例，其中16例为严重脓毒症。选取20例术后SIRS患者和15例健康儿童作为对照组。采用实时定量PCR（qRT-PCR）检测血浆miR-21、miR-125b、miR-132、miR-146a、miR-155和miR-223的表达水平。通过受试者工作特征曲线（ROC）评估miRNA、PCT和CRP对脓毒症的预测价值。采用细胞计数微珠阵列（CBA）检测血浆中TNF-α和IL-10水平。三组间正态分布的定量资料采用方差分析，两组间采用LSD-t检验。对于非正态分布的数据，三组间的多重比较采用Kruskal-Wallis H检验，两组间的差异采用Mann-Whitney U检验进行事后分析。
各组年龄和性别差异无统计学意义。各组血浆中miR-21、miR-125b、miR-132和miR-155的表达差异无统计学意义（均P>0.05）。与SIRS组和对照组相比，脓毒症组miR-146a和miR-223水平上调[（5.7±3.5）×10⁻⁵ vs.（2.4±1.6）×10⁻⁵和（2.6±1.2）×10⁻⁵，（12.5±7.7）×10⁻⁴ vs.（8.3±3.4）×10⁻⁴和（5.3±2.2）×10⁻⁴，均P<0.01]，SIRS组miR-223表达水平高于对照组（P<0.01）。严重脓毒症组miR-146a水平高于一般脓毒症组[（7.1±3.3）×10⁻⁵ vs.（4.6±2.6）×10⁻⁵，P<0.01]。脓毒症组和SIRS组CRP和PCT水平均高于对照组（均P<0.01）。miR-146a、miR-223、PCT和CRP预测脓毒症的ROC曲线下面积（AUC）分别为0.815（95%CI：0.708 - 0.922）、0.678（95%CI：0.537 - 0.818）、0.706（95%CI：0.571 - 0.842）和0.588（95%CI：0.427 - 0.748）。与SIRS组和对照组相比，脓毒症组IL-10和IL-10/TNF-α表达水平上调（均P<0.01）。miR-146a、miR-223与IL-10和IL-10/TNF-α呈正相关（r = 0.545、0.305、0.562、0.373，均P<0.01）。
小儿脓毒症患者血浆中miR-146a和miR-223表达水平显著上调，且与IL-10和IL-10/TNF-α呈正相关，可作为早期诊断标志物，在一定程度上反映病情严重程度。
