目的 分析血清中microRNA 20a与小儿肺炎炎症反应进展的相关性,为临床治疗提供依据。方法 筛选2015年3月－2017年2月治疗的90例诊断为“肺炎”的患儿设为小儿肺炎组(n=90),另选同期健康体检儿童90例设为对照组(n=90)。儿童入院后均分别进行血常规和血沉等的检查;用酶联免疫吸附测定法(ELISA)识别肺炎的特异性抗体;采用全自动特定蛋白分析仪和免疫比浊法对血清中C反应蛋白(CRP)和类风湿因子(RF)浓度进行定量检测;使用TRIzol试剂分离总RNA,按照Prime-Script RT试剂盒说明将血清中microRNA 20a合成逆转录cDNA;通过10％SDS-PAGE凝胶电泳分离蛋白质提取物,并电泳转移到聚偏二氟乙烯膜上进行血清中microRNA 20a蛋白的表达,采用Kolmogorov-Smirnov检验分析血清中microRNA 20a表达与小儿肺炎的炎症反应的相关性。结果 与对照组相比,小儿肺炎组血清中血清免疫球蛋白IgG、IgA水平升高,IgM无显著变化。特定蛋白分析显示小儿肺炎组血清CRP、RF和IL-17含量,microRNA 20a的表达均高于对照组(P<0.05)。microRNA 20a参数与IL-17水平呈显著正相关(r=0.571,P=0.002)。结论 血清中microRNA 20a表达可能与肺炎炎症的发病过程有关,microRNA 20a将有望成为诊断小儿肺炎的标志物,可作为判断小儿肺炎患者预后不良的指标。
Abstract
Objective To explore the correlation between serum microRNA 20a and the progress of inflammatory response in children with pneumonia,in order to provide reference for clinical treatment.Methods Totally 90 children diagnosed with pneumonia from March 2015 to February 2017 were selected as pneumonia group (n=90) and 90 healthy children were enrolled in control group (n=90).Routine blood test and erythrocyte sedimentation rate were conducted in children after admission,and specific antibodies against pneumonia were identified by ELISA.Serum concentration of C-reactive protein (CRP) and rheumatoid factor (RF) was quantitatively detected by automatic specific protein analyzer and immunoturbidimetry.Total RNA was separated by TRIzol reagent and reverse transcription cDNA was synthesized from microRNA 20a in serum according to Prime-Script RT kit instructions.Then,the protein extracts were separated by 10%SDS-PAGE gel electrophoresis and transferred to PVDF membrane to express microRNA 20a protein in 20a.Finally,Kolmogorov-Smirnov test was used to analyze the correlation between the expression of microRNA 20a in serum and inflammatory response in children with pneumonia. Results Compared with the control group,serum levels of IgG and IgA in children with pneumonia increased,but IgM did not change significantly.Specific protein analysis showed that serum CRP,RF and IL-17 levels and the expression level of microRNA 20a in pneumonia group were higher than those in control group (P<0.05).The correlation analysis between microRNA 20a parameters and IL-17 level showed positive correlation and a steep correlation curve (r=0.571,P＝0.002). Conclusions Serum expression level of microRNA 20a may be related to the pathogenesis of pneumonia inflammation.MicroRNA 20a is promising to be a marker for the diagnosis of pediatric pneumonia and can be used as an indicator for the poor prognosis of pediatric pneumonia patients.
关键词
microRNA 20a /
小儿肺炎 /
炎症反应
Key words
microRNA 20a /
pediatric pneumonia /
inflammatory reactions
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