脓毒症血清miRNA表达谱的初步研究

A preliminary study of serum miRNA expression profile in patients with Sepsis

目的：探讨miRNA在脓毒症患者血清中的表达及临床应用价值。方法收集34例脓毒症患者血清（Sepsis组），以及性别年龄与之相匹配的20名健康体检者（健康对照组）和34例具有全身炎症反应患者的血清（SIRS组），利用芯片技术对其中3例Sepsis患者和3例健康体检者进行miRNA表达谱筛选。从筛选出有表达差异的miRNA中，选取与微生物感染相关的7个miRNA，扩大样品量， qRT？PCR对3组样品进行验证，分析评估差异miRNA的临床应用价值。结果芯片实验结果显示Sepsis组与健康对照组共有36个差异表达的miRNA，其中表达上调的miRNA有16个，表达下调的miRNA有20个。经qRT？PCR扩大样品量验证发现，hsa？miRNA？92a？3p在健康对照组、SIRS组和Sepsis组的表达水平依次呈下调趋势（P＜0.05）。hsa？miRNA？92a？3p在评价Sepsis组和健康对照组的曲线下面积（AUC）为0.965，当最佳临界值为0.637时，敏感度为100%，特异度为82.4%。在评价SIRS组和健康对照组的曲线下面积（AUC）为0.844，当最佳临界值为0.698时，敏感度为90.0%，特异度为79.4%。在鉴别诊断Sepsis和SIRS时hsa？miRNA？92a？3p曲线下面积为0.769，当最佳临界值为0.532时，敏感度为64.7%，特异度为88.2%。结论 Sepsis患者血清中特异表达的hsa？miRNA？92a？3p对Sepsis诊断及鉴别诊断具有潜在应用价值。

Objective To investigate the serum profiles and clinical value of miRNA expression in septic patients. Methods We collected serum samples from 34 patients with sepsis （sepsis group）,20 gender？ and age？matched healthy subjects （control group） and 34 patients with inflammatory response syndrome （SIRS group）. miRNA microarray was used to examine the miRNA expression profile in three septic patienttts and three healthy subjects. From all miRNA with differential expression ,we selected seven miRNAs related to microbial infections,expanded the products,and verified them by using qRT？PCR in the three groups of serum samples. Clinical value of these differentially expressed miRNA was analyzed. Results Microarray showed a total of 36 differentially expressed miRNA between the sepsis group and healthy control group,including 16 with upregulated and 20 with downregulated expression. By qRT？PCR expansion and verification,we noted a decreasing trend of hsa？miRNA？92a？3p down？regulation across the healthy control group,SIRS group and sepsis group （P〈0.05）. The area under curve （AUC） of hsa？miRNA？92a？3p in distinguishing sepsis and healthy control was 0.965. At an optimal cutoff of 0.637,the curve yielded a sensitivity of 100% and a specificity of 82.4%. The AUC of hsa？miRNA？92a？3p in distinguishing SIRS and healthy controls was 0.844. At an optimal cutoff of 0.698,the curve yielded a sensitivity of 90.0%and a specificity of 79.4%. The AUC of hsa？miRNA？92a？3p in distinguishing sepsis and SIRS was 0.769. At an optimal cutoff of 0.532,the curve yielded a sensitivity of 64.7%and a specificity of＆amp;nbsp;88.2%. Conclusion The differentially expressed hsa？miRNA？92a？3p found in serum from septic patients may be potentially useful for diagnosis and differential diagnosis of sepsis.