Recombinant Thrombomodulin on Neutrophil Extracellular Traps in Murine Intestinal Ischemia-Reperfusion
Recombinant Thrombomodulin on Neutrophil Extracellular Traps in Murine Intestinal Ischemia-Reperfusion
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DOI：
10.1097/aln.0000000000002898
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发表时间：
2019-10-01
期刊：
影响因子：
8.8
通讯作者：
Morimura, Naoto
中科院分区：
文献类型：
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作者：
Hayase, Naoki;Doi, Kent;Morimura, Naoto
背景资料：在多器官功能障碍中，影响一个器官的损伤会远程影响其他器官，并且受损器官会协同恶化结果。最近，几种介质，包括细胞外组蛋白和中性粒细胞胞外陷阱，被确定为远端器官损伤的贡献者。本研究旨在阐明这些介质是否在肠缺血-再灌注引起的远隔器官损伤中起关键作用。本研究还旨在评估重组血栓调节蛋白的保护作用，据报道，以中和细胞外组蛋白，对多器官功能障碍后肠缺血reperfusion.Methods：肠缺血诱导雄性C57 BL/6 J小鼠通过夹闭的上级肠系膜动脉。重组血栓调节蛋白（10 mg/kg）腹腔内给药，开始再灌注。小鼠进行生存分析，组织学损伤评分，定量聚合酶链反应分析肿瘤坏死因子-α和角化细胞衍生的趋化因子的表达，伊文思蓝染料血管通透性测定，和酶联免疫吸附试验分析组蛋白在空肠，肝脏，肺，和肾脏缺血30或45分钟后。结果：重组血栓调节蛋白产生45分钟缺血后的生存统计学显着改善（缺血再灌注与10 mg/kg重组血栓调节蛋白：0%对33%，n = 21每组，P = 0.001）。重组血栓调节蛋白降低了组织学损伤评分，肿瘤坏死因子-α和角质形成细胞衍生的趋化因子的表达，以及埃文斯蓝染料的外渗，这是增强了30分钟的缺血再灌注，在肝脏中，但不是在肠。缺血再灌注30 min小鼠肝、肠组织中可见组蛋白和中性粒细胞胞外陷阱的聚集。重组血栓调节蛋白减少这些积累，只有在liver.Conclusions：重组血栓调节蛋白改善了雄性小鼠肠缺血再灌注损伤的生存。这些发现表明，组蛋白和中性粒细胞胞外陷阱积累加剧肠缺血再灌注后的远端肝损伤。重组血栓调节蛋白可抑制这些积聚并减轻肝损伤。
Background: In multiple-organ dysfunction, an injury affecting one organ remotely impacts others, and the injured organs synergistically worsen outcomes. Recently, several mediators, including extracellular histones and neutrophil extracellular traps, were identified as contributors to distant organ damage. This study aimed to elucidate whether these mediators play a crucial role in remote organ damage induced by intestinal ischemia-reperfusion. This study also aimed to evaluate the protective effects of recombinant thrombomodulin, which has been reported to neutralize extracellular histones, on multiple-organ dysfunction after intestinal ischemia-reperfusion.Methods: Intestinal ischemia was induced in male C57BL/6J mice via clamping of the superior mesenteric artery. Recombinant thrombomodulin (10 mg/kg) was administered intraperitoneally with the initiation of reperfusion. The mice were subjected to a survival analysis, histologic injury scoring, quantitative polymerase chain reaction analysis of tumor necrosis factor-alpha and keratinocyte-derived chemokine expression, Evans blue dye vascular permeability assay, and enzyme-linked immunosorbent assay analysis of histones in the jejunum, liver, lung, and kidney after 30- or 45-min ischemia. Neutrophil extracellular trap formation was evaluated by immunofluorescence staining.Results: Recombinant thrombomodulin yielded statistically significant improvements in survival after 45-min ischemia (ischemia-reperfusion without vs. with 10 mg/kg recombinant thrombomodulin: 0% vs. 33%, n = 21 per group, P = 0.001). Recombinant thrombomodulin reduced the histologic injury score, expression of tumor necrosis factor-alpha and keratinocyte-derived chemokine, and extravasation of Evans blue dye, which were augmented by 30- min ischemia-reperfusion, in the liver, but not in the intestine. Accumulated histones and neutrophil extracellular traps were found in the livers and intestines of 30-min ischemia-reperfusion-injured mice. Recombinant thrombomodulin reduced these accumulations only in the liver.Conclusions: Recombinant thrombomodulin improved the survival of male mice with intestinal ischemia-reperfusion injury. These findings suggest that histone and neutrophil extracellular trap accumulation exacerbate remote liver injury after intestinal ischemia-reperfusion. Recombinant thrombomodulin may suppress these accumulations and attenuate liver injury.