Mechanical ventilation enhances HMGB1 expression in an LPS-induced lung injury model.
Mechanical ventilation enhances HMGB1 expression in an LPS-induced lung injury model.
复制标题
机械通气增强 LPS 诱导的肺损伤模型中 HMGB1 的表达。
DOI：
10.1371/journal.pone.0074633
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
She S
中科院分区：
文献类型：
--
作者：
Ding N;Wang F;Xiao H;Xu L;She S
机械通气（MV）可增强脂多糖（LPS）攻击肺的炎症反应。高迁移率族蛋白B1（HMGB1）是呼吸机相关性肺损伤的促炎介质，但其机制尚不清楚。本研究探讨了HMGB1在MV和LPS联合治疗的肺部炎症反应中的作用。将48只雄性Sprague-Dawley大鼠随机分为4组：假手术对照组、LPS治疗组、机械通气组、机械通气+LPS治疗组。机械通气动物接受10 ml/kg潮气量，速率为40次呼吸/min，持续4 h。在HMGB1阻断研究中，将16只大鼠随机分配到HMGB1抗体组或对照抗体组，并如上所述使动物经受MV + LPS。A549细胞与不同的信号抑制剂预孵育，然后进行4小时的循环拉伸。检测肺湿/干重（W/D）、支气管肺泡灌洗液（BALF）中总蛋白和IgG浓度、中性粒细胞计数及肺组织学改变。ELISA法检测BALF中白细胞介素1 β（IL-1 β）、IL-6、肿瘤坏死因子-α（TNF-α）、巨噬细胞炎性蛋白2（MIP-2）和高迁移率族蛋白1（HMGB1）的水平。实时定量PCR和Western blot检测HMGB1 mRNA和蛋白表达。Western blot检测I κ B-α、NF-κ B、JNK、ERK和p38的活化。MV可显著增加LPS诱导的肺损伤和HMGB1表达，且与BALF中IL-1 β、IL-6和MIP-2水平升高相关。在体内，气管内给予HMGB1抗体显著减轻肺部炎症损伤。体外实验表明，牵张可通过p38和NF-κ B等信号通路诱导HMGB1的表达。结果表明，中等潮气量MV通过上调HMGB1增强LPS诱导的肺损伤。HMGB1介导肺损伤的机制可能是通过p38和NF-κ B信号通路。
Mechanical ventilation (MV) can augment inflammatory response in lipopolysaccharide (LPS) challenged lungs. High mobility group box 1 protein (HMGB1) is a pro-inflammatory mediator in ventilator-induced lung injury, but its mechanisms are not well defined. This study investigated the role of HMGB1 in lung inflammation in response to the combination of MV and LPS treatment. Forty-eight male Sprague-Dawley rats were randomized to one of four groups: sham control; LPS treatment; mechanical ventilation; mechanical ventilation with LPS treatment. Mechanically ventilated animals received 10 ml/kg tidal volumes at a rate of 40 breaths/min for 4 h. In the HMGB1-blockade study, sixteen rats were randomly assigned to HMGB1 antibody group or control antibody group and animals were subjected to MV+LPS as described above. A549 cells were pre-incubated with different signal inhibitors before subjected to 4 h of cyclic stretch. Lung wet/dry weight (W/D) ratio, total protein and IgG concentration, number of neutrophils in bronchoalveolar lavage fluid (BALF), and lung histological changes were examined. The levels of interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α), macrophage inflammatory protein-2 (MIP-2) and HMGB1 in BALF were measured using ELISA. Real-time quantitative PCR and Western blot were used to analyze mRNA and protein expression of HMGB1. Western blot were employed to analyze the activation of IκB-α, NF-κB, JNK, ERK, and p38. MV significantly augmented LPS-induced lung injury and HMGB1 expression, which was correlated with the increase in IL-1β, IL-6 and MIP-2 levels in BALF. In vivo, intratracheally administration of HMGB1 antibody significantly attenuated pulmonary inflammatory injury. In vitro experiments showed cyclic stretch induced HMGB1 expression through signaling pathways including p38 and NF-κB. The findings indicated that moderate tidal volume MV augmented LPS induced lung injury by up-regulating HMGB1. The mechanism of HMGB1-mediated lung injury is likely to be signaling through p38 and NF-κB pathways.