The β3/5 Integrin-MMP9 Axis Regulates Pulmonary Inflammatory Response and Endothelial Leakage in Acute Lung Injury.
The β3/5 Integrin-MMP9 Axis Regulates Pulmonary Inflammatory Response and Endothelial Leakage in Acute Lung Injury.
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β3/5 整合素-MMP9 轴调节急性肺损伤中的肺部炎症反应和内皮渗漏
DOI：
10.2147/jir.s331939
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发表时间：
2021
影响因子：
4.5
通讯作者：
Luo Y
中科院分区：
文献类型：
--
作者：
Tong Y;Bao C;Xu YQ;Tao L;Zhou Y;Zhuang L;Meng Y;Zhang H;Xue J;Wang W;Zhang L;Pan Q;Shao Z;Hu T;Guo Q;Xue Q;Lu H;Luo Y
背景 急性肺损伤（ALI）是一种严重的呼吸道疾病，发病率和死亡率很高。许多内源性或外源性介质参与 ALI 的病理生理学。在这里，我们发现整合素和基质金属蛋白酶作为过度炎症和内皮通透性的关键决定因素参与 ALI 的调节。方法通过定量实时PCR测定炎症细胞因子的mRNA水平和ELISA测定分泌水平。通过罗丹明B异硫氰酸酯-葡聚糖的通过来检测内皮通透性测定。通过伊文思蓝白蛋白（EBA）测定小鼠肺通透性。蛋白质印迹用于蛋白质水平测量。使用细胞渗透性探针 DCFH-DA 评估细胞内活性氧 (ROS)。小鼠气管内注射脂多糖（LPS）建立肺损伤模型。结果 外源性MMP-9显着加重LPS处理的小鼠肺微血管内皮细胞（PMVEC）的炎症反应和通透性，而敲除MMP-9则表现出相反的表型。在 LPS 处理的 PMVEC 中，整合素 β3 或 β5 的敲低可显着下调 MMP-9 的表达，并在存在或不存在外源 MMP-9 的情况下降低炎症反应和通透性。此外，ROS 清除剂损害了 MMP-9 和整合素 β5 的相互作用，进一步减少了联合治疗（LPS 与外源性 MMP-9）的 PMVEC 中促炎细胞因子的产生和内皮渗漏。体内研究显示，外源性 MMP-9 治疗或敲低 β3 整合素可显着降低 ALI 小鼠的存活率。值得注意的是，单独敲除β5整合素对生存没有显着影响，但与抗MMP-9治疗相结合，可通过改善ALI小鼠过度的肺部炎症和通透性来显着提高生存率。结论 这些发现支持 β3/5 整合素-MMP-9 轴作为内源性信号，在调节 ALI 炎症反应和肺泡毛细血管通透性方面发挥关键作用。
Background Acute lung injury (ALI) is a severe respiratory disease with high rates of morbidity and mortality. Many mediators regarding endogenous or exogenous are involved in the pathophysiology of ALI. Here, we have uncovered the involvement of integrins and matrix metalloproteinases, as critical determinants of excessive inflammation and endothelial permeability, in the regulation of ALI. Methods Inflammatory cytokines were measured by quantitative real-time PCR for mRNA levels and ELISA for secretion levels. Endothelial permeability assay was detected by the passage of rhodamine B isothiocyanate-dextran. Mice lung permeability was assayed by Evans blue albumin (EBA). Western blot was used for protein level measurements. The intracellular reactive oxygen species (ROS) were evaluated using a cell-permeable probe, DCFH-DA. Intratracheal injection of lipopolysaccharide (LPS) into mice was conducted to establish the lung injury model. Results Exogenous MMP-9 significantly aggravated the inflammatory response and permeability in mouse pulmonary microvascular endothelial cells (PMVECs) treated by LPS, whereas knockdown of MMP-9 exhibited the opposite phenotypes. Knockdown of integrin β3 or β5 in LPS-treated PMVECs significantly downregulated MMP-9 expression and decreased inflammatory response and permeability in the presence or absence of exogenous MMP-9. Additionally, the interaction of MMP-9 and integrin β5 was impaired by a ROS scavenger, which further decreased the pro-inflammatory cytokines production and endothelial leakage in PMVECs subjected to co-treatment (LPS with exogenous MMP-9). In vivo studies, exogenous MMP-9 treatment or knockdown β3 integrin significantly decreased survival in ALI mice. Notably, knockdown of β5 integrin alone had no remarkable effect on survival, but which combined with anti-MMP-9 treatment significantly improved the survival by ameliorating excessive lung inflammation and permeability in ALI mice. Conclusion These findings support the β3/5 integrin-MMP-9 axis as an endogenous signal that could play a pivotal role in regulating inflammatory response and alveolar-capillary permeability in ALI.