Wang Yahui, Wu Yanqi, Liu Bo, Yang Huilin, Qian Hong, Cheng Yumei, Li Xiang, Yang Guixia, Zheng Xinghao, Shen Feng
Department of Intensive Care Unit, Guizhou Medical University Affiliated Hospital Guiyang 550001, Guizhou, China.
Department of Intensive Care Unit, The People's Hospital of Weining County Weining County 553100, Guizhou, China.
Am J Transl Res. 2022 Jun 15;14(6):3854-3863. eCollection 2022.
Alveolar hypercoagulation and fibrinolytic inhibition are shown to be associated with refractory hypoxemia in acute respiratory distress syndrome (ARDS), and the NF-κB pathway is involved in this process. The purpose of this study is to explore the role of NEMO-binding domain peptide (NBDP) in alleviating alveolar hypercoagulation and fibrinolytic inhibition induced by lipopolysaccharide (LPS) in ARDS mice and its related mechanisms.
ARDS was induced by inhalation of LPS (mg/L) in adult male BALB/c mice. Mice were treated with intratracheal inhalation of NBDP or saline aerosol at increased concentrations 30 minutes before LPS administration. Six hours after LPS treatment, bronchoalveolar lavage fluids (BALF) were collected and then all mice were euthanized. In addition, coagulation and fibrinolysis associated factors in lung tissues and BALF were detected, and the activation of NF-κB signaling pathway was observed.
NBDP pretreatment dose-dependently inhibited the expression of tissue factor (TF) and plasminogen activator inhibitor (PAI) 1 in lung tissues, reduced the secretions of TF, PAI-1, thrombin-antithrombin (TAT) complex, and promoted activated protein C (APC) secretion in BALF induced by LPS. LPS-induced high expression of pulmonary procollagen peptide type lll (PIIIP) was also reduced in a dose-dependent manner under NBDP pretreatment. Western blotting showed that NBDP pretreatment significantly attenuated LPS-induced activation of IKKα/β, Iκα and NF-κB p65. NBDP pretreatment also inhibited the DNA binding activity of p65 induced by LPS. We also noticed that NBDP protected mice against LPS-induced lung injury in a dose-dependent manner.
The experimental findings demonstrate that through inhibiting the NF-κB signaling pathway, NBDP dose-dependently ameliorates LPS-induced alveolar hypercoagulation and fibrinolytic inhibition, which is expected to be a new therapeutic target to correct the abnormalities of alveolar coagulation and fibrinolytic pathways in ARDS.
肺泡高凝状态和纤溶抑制与急性呼吸窘迫综合征（ARDS）中的难治性低氧血症相关，且核因子κB（NF-κB）信号通路参与此过程。本研究旨在探讨核因子κB必需调节蛋白结合域肽（NBDP）在减轻ARDS小鼠中脂多糖（LPS）诱导的肺泡高凝状态和纤溶抑制中的作用及其相关机制。
通过吸入LPS（mg/L）诱导成年雄性BALB/c小鼠发生ARDS。在给予LPS前30分钟，对小鼠进行气管内吸入不同浓度的NBDP或盐雾气溶胶处理。LPS处理6小时后，收集支气管肺泡灌洗液（BALF），然后对所有小鼠实施安乐死。此外，检测肺组织和BALF中与凝血和纤溶相关的因子，并观察NF-κB信号通路的激活情况。
NBDP预处理剂量依赖性地抑制肺组织中组织因子（TF）和纤溶酶原激活物抑制剂（PAI）1的表达，减少LPS诱导的BALF中TF、PAI-1、凝血酶-抗凝血酶（TAT）复合物的分泌，并促进活化蛋白C（APC）的分泌。在NBDP预处理下，LPS诱导的肺Ⅲ型前胶原肽（PIIIP）高表达也呈剂量依赖性降低。蛋白质免疫印迹法显示，NBDP预处理显著减弱LPS诱导的IKKα/β、Iκa和NF-κB p65的激活。NBDP预处理还抑制LPS诱导的p65的DNA结合活性。我们还注意到，NBDP以剂量依赖性方式保护小鼠免受LPS诱导的肺损伤。
实验结果表明，NBDP通过抑制NF-κB信号通路，剂量依赖性地改善LPS诱导的肺泡高凝状态和纤溶抑制，有望成为纠正ARDS中肺泡凝血和纤溶途径异常的新治疗靶点。
