Treutiger C J, Mullins G E, Johansson A-S M, Rouhiainen A, Rauvala H M E, Erlandsson-Harris H, Andersson U, Yang H, Tracey K J, Andersson J, Palmblad J E W
Department of Medicine, Center for Infectious Medicine, Karolinska Institute at Huddinge University Hospital, Stockholm, Sweden.
J Intern Med. 2003 Oct;254(4):375-85. doi: 10.1046/j.1365-2796.2003.01204.x.
Severe sepsis and septic shock is a consequence of a generalized inflammatory systemic response because of an invasive infection that may result in acute organ dysfunction. Mortality is high despite access to modern intensive care units. The nuclear DNA binding protein high mobility group 1 (HMGB1) protein has recently been suggested to act as a late mediator of septic shock via its function as a macrophage-derived pro-inflammatory cytokine (J Exp Med 2000; 192: 565, Science1999; 285: 248). We investigated the pro-inflammatory activities of the A-box and the B-box of HMGB1 on human umbilical venular endothelial cells (HUVEC).
The HUVEC obtained from healthy donors were used for experiments. Recombinant human full-length HMGB1, A-box and B-box were cloned by polymerase chain reaction (PCR) amplification from a human brain quick-clone cDNA. The activation of HUVEC was studied regarding (i) upregulation of adhesion molecules, (ii) the release of cytokines and chemokines, (iii) the adhesion of neutrophils to HUVEC, (iv) the activation of signalling transduction pathways and (v) the involvement of the receptor for advanced glycation end-products (RAGE).
The full-length protein and the B-box of HMGB1 dose-dependently activate HUVEC to upregulate adhesion molecules such as ICAM-1, VCAM-1 and E-selectin and to release IL-8 and G-CSF. The activation of HUVEC could be inhibited to 50% by antibodies directed towards the RAGE. HMGB1-mediated HUVEC stimulation resulted in phosphorylation of the ELK-1 signal transduction protein and a nuclear translocation of p65 plus c-Rel, suggesting that HMGB1 signalling is regulated in endothelial cells through NF-kappaB.
The HMGB1 acts as a potent pro-inflammatory cytokine on HUVEC and the activity is mainly mediated through the B-box of the protein. HMGB1 may be a key factor mediating part of the pro-inflammatory response occurring in septic shock and severe inflammation.
严重脓毒症和脓毒性休克是侵袭性感染引起的全身性炎症反应的后果，可能导致急性器官功能障碍。尽管有现代重症监护病房，死亡率仍很高。近来有人提出，核DNA结合蛋白高迁移率族蛋白1（HMGB1）通过其作为巨噬细胞衍生的促炎细胞因子的功能，充当脓毒性休克的晚期介质（《实验医学杂志》2000年；192：565，《科学》1999年；285：248）。我们研究了HMGB1的A盒和B盒对人脐静脉内皮细胞（HUVEC）的促炎活性。
使用从健康供体获得的HUVEC进行实验。通过聚合酶链反应（PCR）扩增从人脑快速克隆cDNA中克隆重组人全长HMGB1、A盒和B盒。关于以下方面研究了HUVEC的激活：（i）黏附分子的上调，（ii）细胞因子和趋化因子的释放，（iii）中性粒细胞与HUVEC的黏附，（iv）信号转导途径的激活，以及（v）晚期糖基化终产物受体（RAGE）的参与。
HMGB1的全长蛋白和B盒以剂量依赖方式激活HUVEC，上调细胞间黏附分子-1（ICAM-1）、血管细胞黏附分子-1（VCAM-1）和E-选择素等黏附分子，并释放白细胞介素-8（IL-8）和粒细胞集落刺激因子（G-CSF）。针对RAGE的抗体可将HUVEC的激活抑制50%。HMGB1介导的HUVEC刺激导致ELK-1信号转导蛋白磷酸化以及p65和c-Rel的核转位，表明HMGB1信号在内皮细胞中通过核因子κB（NF-κB）调节。
HMGB1对HUVEC起强效促炎细胞因子的作用，其活性主要通过该蛋白的B盒介导。HMGB1可能是介导脓毒性休克和严重炎症中部分促炎反应的关键因素。
