α-烯醇化酶通过蛋白酶激活受体2的纤溶酶激活引起肺微血管内皮细胞的促炎激活并使中性粒细胞致敏。
α-Enolase Causes Proinflammatory Activation of Pulmonary Microvascular Endothelial Cells and Primes Neutrophils Through Plasmin Activation of Protease-Activated Receptor 2.
作者信息
Bock Ashley, Tucker Nicole, Kelher Marguerite R, Khan Samina Y, Gonzalez Eduardo, Wohlauer Max, Hansen Kirk, Dzieciatkowska Monika, Sauaia Angels, Banerjee Anirban, Moore Ernest E, Silliman Christopher C
机构信息
*Research Laboratory Bonfils Blood Center and †School of Medicine, University of Colorado Denver; Departments of ‡Surgery, §Pediatrics, and ∥Cell Biology, School of Medicine, University of Colorado Denver, Aurora; and ¶School of Public Health, University of Colorado; and **Department of Surgery Denver Health Medical Center, Denver, Colorado.
出版信息
Shock. 2015 Aug;44(2):137-42. doi: 10.1097/SHK.0000000000000394.
UNLABELLED
Proinflammatory activation of vascular endothelium leading to increased surface expression of adhesion molecules and neutrophil (PMN) sequestration and subsequent activation is paramount in the development of acute lung injury and organ injury in injured patients. We hypothesize that α-enolase, which accumulates in injured patients, primes PMNs and causes proinflammatory activation of endothelial cells leading to PMN-mediated cytotoxicity.
METHODS
Proteomic analyses of field plasma samples from injured versus healthy patients were used for protein identification. Human pulmonary microvascular endothelial cells (HMVECs) were incubated with α-enolase or thrombin, and intercellular adhesion molecule-1 surface expression was measured by flow cytometry. A two-event in vitro model of PMN cytotoxicity HMVECs activated with α-enolase, thrombin, or buffer was used as targets for lysophosphatidylcholine-primed or buffer-treated PMNs. The PMN priming activity of α-enolase was completed, and lysates from both PMNs and HMVECs were immunoblotted for protease-activated receptor 1 (PAR-1) and PAR-2 and coprecipitation of α-enolase with PAR-2 and plasminogen/plasmin.
RESULTS
α-Enolase increased 10.8-fold in injured patients (P < 0.05). Thrombin and α-enolase significantly increased intercellular adhesion molecule-1 surface expression on HMVECs, which was inhibited by antiproteases, induced PMN adherence, and served as the first event in the two-event model of PMN cytotoxicity. α-Enolase coprecipitated with PAR-2 and plasminogen/plasmin on HMVECs and PMNs and induced PMN priming, which was inhibited by tranexamic acid, and enzymatic activity was not required.
CONCLUSIONS
α-Enolase increases after injury and may activate pulmonary endothelial cells and prime PMNs through plasmin activity and PAR-2 activation. Such proinflammatory endothelial activation may predispose to PMN-mediated organ injury.
未标记
血管内皮的促炎激活导致黏附分子表面表达增加、中性粒细胞（PMN）滞留及随后的激活，这在受伤患者急性肺损伤和器官损伤的发展中至关重要。我们推测，在受伤患者体内积聚的α-烯醇化酶会使PMN致敏，并导致内皮细胞的促炎激活，从而导致PMN介导的细胞毒性。
方法
对受伤患者与健康患者的现场血浆样本进行蛋白质组学分析以鉴定蛋白质。将人肺微血管内皮细胞（HMVECs）与α-烯醇化酶或凝血酶孵育，通过流式细胞术测量细胞间黏附分子-1的表面表达。使用α-烯醇化酶、凝血酶或缓冲液激活的HMVECs作为溶血磷脂酰胆碱致敏或缓冲液处理的PMN的靶标，建立PMN细胞毒性的双事件体外模型。完成了α-烯醇化酶的PMN致敏活性研究，并对PMN和HMVECs的裂解物进行免疫印迹分析，检测蛋白酶激活受体1（PAR-1）和PAR-2以及α-烯醇化酶与PAR-2和纤溶酶原/纤溶酶的共沉淀。
结果
受伤患者体内α-烯醇化酶增加了10.8倍（P < 0.05）。凝血酶和α-烯醇化酶显著增加了HMVECs上细胞间黏附分子-1的表面表达，这被抗蛋白酶抑制，诱导了PMN黏附，并在PMN细胞毒性的双事件模型中作为第一个事件。α-烯醇化酶在HMVECs和PMN上与PAR-2和纤溶酶原/纤溶酶共沉淀，并诱导PMN致敏，这被氨甲环酸抑制，且不需要酶活性。
结论
受伤后α-烯醇化酶增加，可能通过纤溶酶活性和PAR-2激活来激活肺内皮细胞并使PMN致敏。这种促炎内皮激活可能易导致PMN介导的器官损伤。
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