Markers of neutrophil activation and extracellular traps formation are predictive of appendicitis in mice and humans: a pilot study.
Markers of neutrophil activation and extracellular traps formation are predictive of appendicitis in mice and humans: a pilot study.
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中性粒细胞活化和细胞外陷阱的标志物预测了小鼠和人类的阑尾炎：一项试点研究。
DOI：
10.1038/s41598-020-74370-9
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发表时间：
2020-10-26
影响因子：
4.6
通讯作者：
Klinke M
中科院分区：
文献类型：
--
作者：
Boettcher M;Esser M;Trah J;Klohs S;Mokhaberi N;Wenskus J;Trochimiuk M;Appl B;Reinshagen K;Raluy LP;Klinke M
阑尾炎是小儿外科最常见的急症之一，但目前用于诊断的生物标志物不具有特异性且预测价值低。由于嗜中性粒细胞和细胞外陷阱（NET）是抵抗细菌感染的免疫防御的重要组成部分，并且阑尾炎被认为是阑尾的炎症反应，因此我们假设嗜中性粒细胞活化和NET形成在阑尾炎的发展和维持中起重要作用。因此，本初步研究旨在建立小鼠阑尾炎模型，并评估用于诊断小鼠和人类阑尾炎的标记物。该研究使用了20只（12只阑尾炎小鼠和8只对照小鼠）6周龄小鼠，这些小鼠使用改良的盲肠结扎穿刺程序进行晚期阑尾炎诱导。在研究期间，评估了游离DNA、中性粒细胞弹性蛋白酶（NE）、髓过氧化物酶（MPO）和瓜氨酸化组蛋白H3（H3cit）。此外，对5例经组织学证实的阑尾炎患儿和5例匹配的卡他性阑尾炎对照组的样本进行了相同生物标志物的检查。此外，NE，MPO和H3cit通过免疫荧光在小鼠和人类中进行组织学评估。阑尾炎组中的所有小鼠均发展为晚期阑尾炎伴局灶性腹膜炎。在患有阑尾炎的小鼠和人类中，与对照相比，血液和组织中的中性粒细胞活化和炎症形成的标志物（尤其是cfDNA、NE和H3cit）显著升高。最终，生物标志物与组织表达以及疾病严重程度非常相关。似乎中性粒细胞活化和可能的NET有助于阑尾炎的发展，中性粒细胞活化和ET形成的生物标志物反映疾病的严重程度，因此可以用作阑尾炎的生物标志物。然而，还需要大量的前瞻性临床研究来证实我们的发现。
Appendicitis is one of the most frequent emergencies in pediatric surgery, yet current biomarkers for diagnosis are unspecific and have low predictive values. As neutrophils and extracellular traps (ETs) are an essential component of the immune defense against bacterial infections, and appendicitis is considered an inflammation reaction of the appendix, we hypothesized that neutrophil activation and NET formation play an essential role in appendicitis development and maintenance. Therefore, this pilot study aimed to establish a murine model of appendicitis and to evaluate ETs markers to diagnose appendicitis in mice and humans. The study used 20 (12 appendicitis- and 8 controls) 6-week old mice which underwent advanced appendicitis induction using a modified caecal ligation puncture procedure. During the study, cell-free DNA, neutrophil elastase (NE), myeloperoxidase (MPO), and citrullinated Histone H3 (H3cit) were assessed. Additionally, samples of 5 children with histologically confirmed appendicitis and 5 matched controls with catarrhal appendicitis, were examined for the same biomarkers. Moreover, NE, MPO, and H3cit were assessed histologically via immunofluorescence in mice and humans. All mice in the appendicitis group developed an advanced form of appendicitis with focal peritonitis. In mice and humans with appendicitis, markers of neutrophil activation and ETs formation (especially cfDNA, NE and H3cit) were significantly elevated in blood and tissue compared to controls. Ultimately, biomarkers correlated extremely well with tissue expression and thus disease severity. It appears that neutrophil activation and possibly NETs contribute to appendicitis development and biomarkers of neutrophil activation and ET formation reflect disease severity and thus could be used as biomarkers for appendicitis. However, large prospective clinical studies are needed to confirm our findings.