目的：观察白虎人参汤联合复方薤白胶囊治疗前后重症肺部感染患者外周血特异性血清微小RNA（miRNA，miR）表达谱的差异性。方法选择2012年1月至2013年12月江苏大学附属昆山市第一人民医院呼吸科住院的重症肺部感染痰热腑实证患者136例，在基础治疗的同时加用白虎人参汤〔石膏30 g（打碎，绵裹，先下），西洋参4 g，浙贝母10 g，柴胡10 g，黄芩10 g，炒山栀10 g，连翘12 g，金银花10 g，鸭跖草15 g，炙甘草4 g〕联合复方薤白胶囊（主要由薤白12 g，瓜蒌12 g，半夏10 g，黄连3 g组成）治疗。用Solexa平台测序分析比较治疗前后患者外周血miRNA表达谱变化，实时荧光定量-聚合酶链反应（qRT-PCR）验证外周血特异性miRNA。绘制治疗前后重症肺部感染患者的受试者工作特征曲线（ROC曲线），利用ROC曲线分析血清miRNA对联合治疗疗效的敏感度和特异度，评估其临床价值。结果 Solexa平台测序检测提示，加用白虎人参汤联合复方薤白胶囊治疗前后共42种miRNA出现2倍以上的差异表达；qRT-PCR表明miR-146a、miR-125b和miR-155表达显著下降超过10倍，提示其可能是特异性miRNA。其中：miR-146a的ROC曲线下面积（AUC）＝0.81，95%CI＝0.73～0.89；miR-125b的AUC＝0.77，95%可信区间（95%CI）＝0.71～0.83；miR-155的AUC＝0.89，95%CI＝0.83～0.95。通过选定的临床截断（cut off）值，对3种血清miRNA的敏感度和特异度进行计算，miR-146a为63%和87%，miR-125b为53%和73%，miR-155为56%和83%。结论加用白虎人参汤联合复方薤白胶囊治疗重症肺炎可使患者外周血miRNA表达谱发生显著变化，其中miR-146a、miR-125b和miR-155可能是其特异性改变miRNA。%ObjectiveTo study the differences in expression profiles of peripheral blood specific serum micro RNA(miRNA) in the comparison between those before and after Baihu Renshen decoction combined with Fufang Xiebai capsules for treatment of patients with severe pneumonia.Methods 136 patients with severe pulmonary infection accompanied by heat-phlegmand sthenic-fu syndrome admitted in the Department of Respiratory Medicine of Kunshan First People's Hospital Affiliated to Jiangsu University from January 2012 to December 2013 were enrolled in the study. They were treated with basic treatment and Baihu Renshen decoction〔gypsum 30 g(broken, wrapped in cotton cloth, firstly put in water to be boiled), American ginseng 4 g, fritillaria 10 g, bupleurum chinense 10 g, skullcap 10 g, fried gardenia(pittosporm root) 10 g, forsythia 12 g, honeysuckle 10 g, spiderwort grass(dayflower) 15 g, prepared licorice root 4 g〕 combined with Fufang Xiebai capsules〔mainly composed of longstamen onion bulb(macrostem onion)12 g, melon wilt(fructus trichosanthis) 12 g, pinellia 10 g, coptis chinensis 3 g〕. The changes of serum miRNA expression profiles before and after treatment were analyzed by Solexa sequencing. The peripheral blood serum specific miRNAs were confirmed by real-time fluorescent quantitation-polymerase chain reaction(qRT-PCR). Before and after treatment, the receiver operating characteristic curve(ROC curve) of patients with severe pulmonary infection was drawn, and the ROC curve was used to analyze the sensitivity and specificity of serum miRNA to the efficacy of the combination therapy and evaluate its clinical value.Results Solexa sequencing suggested that 42 kinds of miRNA appear to have over 2-fold in difference in the comparison between profiles before and after Baihu Renshen decoction combined with Fufang Xiebai capsules treatment. Real-time qRT-PCR confirmed miR-146a, miR-125b and miR-155 expressions were significantly decreased over 10 fold, suggesting that they should be specific miRNAs. The area under the ROC curve(AUC) of miR-146a was 0.81,95% confidence interval(95%CI)=0.73 - 0.89,miR-125b AUC=0.77,95%CI=0.71 - 0.83 and miR-155 AUC=0.89, 95%CI=0.83 - 0.95. Then, through selected clinical truncated(cut off)value,the sensitivity and specificity of three kinds of serum miRNA were calculated: 63% and 87% of miR-146a, 53% and 73% of miR-125b and 56% and 83% of miR-155.ConclusionAddition of Baihu Renshen decoction combined with Fufang Xiebai capsules on the basic treatment can induce significant changes of peripheral blood serum miRNA expression profiles in patients with severe pneumonia, and miR-146a, miR-125b and miR-155 among miRNAs could be their specific changes.
展开▼
