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Histones released during NETs formation damage endothelial glycocalyx aggravating pathogenesis of sepsis

NETs形成过程中释放的组蛋白损伤内皮糖萼，加重脓毒症的发病机制

基本信息

批准号：
18K16543
负责人：
Shrestha Binita
金额：
$ 2.66万
依托单位：
Kagoshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Early-Career Scientists
财政年份：
2018
资助国家：
日本
起止时间：
2018-04-01 至 2019-03-31
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-18K16543/
关键词：
Glycocalyx Sepsis Histones HIstones NETs

项目摘要

Sepsis is defined as a life-threatening organ dysfunction that is the leading cause of death in critically ill patients in intensive care units. The current sepsis definition focuses on organ dysfunction, which is associated with high mortality. The endothelial glycocalyx is crucial for maintenance of the vascular barrier. Alteration in the composition of the glycocalyx after exposure to an inflammatory insult is one of the earliest features during sepsis. We proposed that histones, released during sepsis, binds with the components of glycocalyx and cause the damage to glycocalyx layer thus causing disruption of the endothelial barrier leading to pathogenesis of sepsis. In our study we treated HUVECs with histones H3 and H4 for 2h. Cells were stained with Wheat Germ Agglutinin (WGA), which binds with sialic acid in glycocalyx layer, to visualize the glycocalyx. At the histones concentration of 20μg/ml, glycocalyx layers seemed to disrupt at some parts of cell membranes. Thus we could show in our preliminary study that histones H3 and H4 at the total concentration of 20μg/ml can disrupt glycocalyx layer of HUVECs. This result should be further confirmed using glycocalyx specific antibodies. Also syndecan level in supernatant should be measured to confirm disruption of the glycocalyx layer. From our study up to now, we can say that histones cause damage to glycocalyx layer.

脓毒症被定义为一种危及生命的器官功能障碍，是重症监护病房危重患者死亡的主要原因。目前对脓毒症的定义主要集中在器官功能障碍，这与高死亡率有关。内皮糖基化对于维持血管屏障起着至关重要的作用。在受到炎性侮辱后，糖萼成分的改变是脓毒症最早的特征之一。我们认为，在脓毒症过程中释放的组蛋白与糖萼组分结合，导致糖帽层的损伤，从而导致内皮屏障的破坏，从而导致脓毒症的发病。在我们的研究中，我们用组蛋白H3和H4处理人脐静脉内皮细胞2小时。细胞用小麦胚凝集素(WGA)染色，该凝集素与糖萼层中的唾液酸结合，以显示糖萼。当组蛋白浓度为20μg/ml时，部分细胞膜上的糖帽层出现断裂。因此，我们的初步研究表明，总浓度为20μg/ml的组蛋白H3和H4可以破坏人脐静脉内皮细胞的糖帽层。这一结果应使用糖萼特异性抗体进一步证实。此外，还应测量上清液中的Syndecan水平，以确认糖帽层的破坏。从我们的研究到目前为止，我们可以说组蛋白对糖帽层造成了损害。