Early diagnosis using canonical discriminant analysis of innate immune receptor gene expression profiles in a murine infectious or sterile systemic inflammation model
Early diagnosis using canonical discriminant analysis of innate immune receptor gene expression profiles in a murine infectious or sterile systemic inflammation model
复制标题
在小鼠感染性或无菌全身性炎症模型中使用先天免疫受体基因表达谱的典型判别分析进行早期诊断
DOI：
10.1097/ta.0000000000001789
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发表时间：
2018
影响因子：
3.4
通讯作者：
Tasaki Osamu
中科院分区：
文献类型：
--
作者：
Tajima Goro;Tokunaga Ayako;Umehara Takahiro;Ikematsu Kazuya;Miyamoto Junya;Sato Shuntaro;Tasaki Osamu
背景：全身性炎症患者的感染很难用单一的生物标志物进行诊断。我们旨在阐明感染性或无菌性炎症中先天免疫受体基因表达谱变化的时间过程，并利用基因表达谱的典型判别分析（CDA）建立一种早期诊断方法。方法采用盲肠结扎穿刺（CLP）和20%全层烧伤（burn）模型，比较感染性炎症和无菌性炎症。C57BL/6小鼠分别接受假治疗（n= 9× 3组）、CLP （n= 12× 3组）和烧伤（n= 12× 3组）。分别于损伤后6、12、24小时处死小鼠，从全血中提取总RNA。采用实时定量聚合酶链反应检测天然免疫受体toll样受体2 （TLR2）、TLR4、TLR9、NLRP3（核苷酸结合寡聚化结构域样受体家族pyrin结构域3）和维甲酸诱导基因i的基因表达。将所有基因表达作为模式进行评估，每个值进行标准化，并在每个时间点进行CDA。结果损伤后6 h， CLP和Burn的TLR2和TLR4基因表达均较sham小鼠显著升高（p< 0.05）。与sham和Burn小鼠相比，CLP中TLR9基因表达在损伤后12小时和24小时显著降低（p< 0.05），但在损伤后6小时无显著降低。损伤后6 h和24 h， CLP和Burn小鼠NLRP3基因表达明显高于sham小鼠（p< 0.05）。在CDA中，各组在损伤后仅6小时显示出不同的基因表达模式。各组分类清晰，各时间点分类错误率均为0%。结论先天性免疫受体基因表达谱的扫描判别分析可作为一种从损伤早期开始诊断复杂全身性炎症病理生理的新方法。
BACKGROUNDInfection in patients with systemic inflammation is difficult to diagnose with a single biomarker. We aimed to clarify the time course of change in the gene expression profile of innate immune receptors in infectious or sterile inflammation and to establish an early diagnostic method using canonical discriminant analysis (CDA) of the gene expression profile.METHODSTo compare infectious and sterile inflammation, we used cecal ligation and puncture (CLP) and 20% full-thickness burn injury (Burn) models. C57BL/6 mice underwent sham treatment (n= 9× three groups), CLP (n= 12× three groups), or Burn (n= 12× three groups) injury. Mice were killed at 6, 12, and 24 hours after injury, and total RNA was extracted from whole blood. We used quantitative real-time polymerase chain reaction to investigate gene expression of innate immune receptors Toll-like receptor 2 (TLR2), TLR4, TLR9, NLRP3 (nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3), and retinoic acid–inducible gene I. To evaluate all gene expression together as patterns, each value was standardized, and CDA was performed at each time point.RESULTSGene expression of TLR2 and TLR4 was already significantly increased in both CLP and Burn compared with sham mice at 6 hours after injury (p< 0.05). Gene expression of TLR9 was significantly decreased in CLP compared with sham and Burn mice at 12 hours and 24 hours after injury (p< 0.05) but not at 6 hours. Gene expression of NLRP3 was significantly increased in CLP and Burn compared with sham mice at 6 hours and 24 hours after injury (p< 0.05). In the CDA, each group showed distinctive gene expression patterns at only 6 hours after injury. Each group was clearly classified, and the classification error rates were 0% at all of the time points.CONCLUSIONSCanonical discriminant analysis of the gene expression profile of innate immune receptors could be a novel approach for diagnosing the pathophysiology of complicated systemic inflammation from the early stage of injury.