人类糖尿病酮症酸中毒血浆中升高的嗜天青颗粒白细胞酶可降解脑血管内皮连接蛋白。

Elevated Leukocyte Azurophilic Enzymes in Human Diabetic Ketoacidosis Plasma Degrade Cerebrovascular Endothelial Junctional Proteins.

作者信息

Woo Martin M H, Patterson Eric K, Clarson Cheril, Cepinskas Gediminas, Bani-Yaghoub Mahmud, Stanimirovic Danica B, Fraser Douglas D

机构信息

1Children's Health Research Institute, London, ON, Canada.2Department of Physiology and Pharmacology, Western University, London, ON, Canada.3Centre for Critical Illness Research, Lawson Health Research Institute, London, ON, Canada.4Department of Translational Bioscience, National Research Council Canada, Ottawa, ON, Canada.5Department of Medicine, Western University, London, ON, Canada.6Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON, Canada.7Department of Paediatrics, Western University, London, ON, Canada.8Department of Clinical Neurological Sciences, Western University, London, ON, Canada.

出版信息

Crit Care Med. 2016 Sep;44(9):e846-53. doi: 10.1097/CCM.0000000000001720.

DOI:10.1097/CCM.0000000000001720

PMID:27071071

Abstract

OBJECTIVE

Diabetic ketoacidosis in children is associated with vasogenic cerebral edema, possibly due to the release of destructive polymorphonuclear neutrophil azurophilic enzymes. Our objectives were to measure plasma azurophilic enzyme levels in children with diabetic ketoacidosis, to correlate plasma azurophilic enzyme levels with diabetic ketoacidosis severity, and to determine whether azurophilic enzymes disrupt the blood-brain barrier in vitro.

DESIGN

Prospective clinical and laboratory study.

SETTING

The Children's Hospital, London Health Sciences Centre.

SUBJECTS

Pediatric type 1 diabetes patients; acute diabetic ketoacidosis or age-/sex-matched insulin-controlled.

MEASUREMENTS AND MAIN RESULTS

Acute diabetic ketoacidosis in children was associated with elevated polymorphonuclear neutrophils. Plasma azurophilic enzymes were elevated in diabetic ketoacidosis patients, including human leukocyte elastase (p < 0.001), proteinase-3 (p < 0.01), and myeloperoxidase (p < 0.001). A leukocyte origin of human leukocyte elastase and proteinase-3 in diabetic ketoacidosis was confirmed with buffy coat quantitative real-time polymerase chain reaction (p < 0.01). Of the three azurophilic enzymes elevated, only proteinase-3 levels correlated with diabetic ketoacidosis severity (p = 0.002). Recombinant proteinase-3 applied to human brain microvascular endothelial cells degraded both the tight junction protein occludin (p < 0.05) and the adherens junction protein VE-cadherin (p < 0.05). Permeability of human brain microvascular endothelial cell monolayers was increased by recombinant proteinase-3 application (p = 0.010).

CONCLUSIONS

Our results indicate that diabetic ketoacidosis is associated with systemic polymorphonuclear neutrophil activation and degranulation. Of all the polymorphonuclear neutrophil azurophilic enzymes examined, only proteinase-3 correlated with diabetic ketoacidosis severity and potently degraded the blood-brain barrier in vitro. Proteinase-3 might mediate vasogenic edema during diabetic ketoacidosis, and selective proteinase-3 antagonists may offer future vascular- and neuroprotection.

摘要

目的

儿童糖尿病酮症酸中毒与血管源性脑水肿有关，可能是由于破坏性多形核中性粒细胞嗜天青酶的释放。我们的目标是测量糖尿病酮症酸中毒患儿的血浆嗜天青酶水平，将血浆嗜天青酶水平与糖尿病酮症酸中毒的严重程度相关联，并确定嗜天青酶在体外是否会破坏血脑屏障。

设计

前瞻性临床和实验室研究。

地点

伦敦健康科学中心儿童医院。

研究对象

儿科1型糖尿病患者；急性糖尿病酮症酸中毒患者或年龄/性别匹配的胰岛素控制患者。

测量和主要结果

儿童急性糖尿病酮症酸中毒与多形核中性粒细胞升高有关。糖尿病酮症酸中毒患者的血浆嗜天青酶升高，包括人白细胞弹性蛋白酶（p<0.001）、蛋白酶-3（p<0.01）和髓过氧化物酶（p<0.001）。通过血沉棕黄层定量实时聚合酶链反应证实糖尿病酮症酸中毒中人白细胞弹性蛋白酶和蛋白酶-3的白细胞来源（p<0.01）。在升高的三种嗜天青酶中，只有蛋白酶-3水平与糖尿病酮症酸中毒严重程度相关（p=0.002）。应用于人脑微血管内皮细胞的重组蛋白酶-3可降解紧密连接蛋白闭合蛋白（p<0.05）和黏附连接蛋白血管内皮钙黏蛋白（p<0.05）。应用重组蛋白酶-3后人脑微血管内皮细胞单层的通透性增加（p=0.010）。