Exosome-delivered syndecan-1 rescues acute lung injury via a FAK/p190RhoGAP/RhoA/ROCK/NF-κB signaling axis and glycocalyx enhancement
Exosome-delivered syndecan-1 rescues acute lung injury via a FAK/p190RhoGAP/RhoA/ROCK/NF-κB signaling axis and glycocalyx enhancement
复制标题
外泌体递送的 syndecan-1 通过 FAK/p190RhoGAP/RhoA/ROCK/NF-kappa B 信号轴和糖萼增强来拯救急性肺损伤
DOI：
10.1016/j.yexcr.2019.111596
复制
发表时间：
2019-11-01
影响因子：
3.7
通讯作者：
Huan, Jingning
中科院分区：
文献类型：
--
作者：
Zhang, Chuankai;Guo, Feng;Huan, Jingning
急性肺损伤（ALI）的特征是富含蛋白质的肺水肿、严重的低氧血症以及促炎细胞因子和细胞的涌入。目前临床上尚无有效的药物疗法。内皮细胞中的 Syndecan-1 具有保护内皮屏障功能并抑制炎症反应的潜力。因此，本研究旨在确定包裹syndecan-1的外泌体是否能够在ALI中取得理想的治疗效果。从慢病毒转染的小鼠肺微血管内皮细胞 (MPMVEC) 的条件培养基中分离外泌体，并通过纳米颗粒跟踪分析 (NTA)、透射电子显微镜 (TEM) 和蛋白质印迹进行表征。通过气管内给予脂多糖（LPS）并用外泌体处理来诱导 ALI 小鼠模型。检查肺水肿、炎症和糖萼厚度。通过 MPMVEC 中的免疫印迹验证了可能的机制。纯化的外泌体包括SDC1-high-Exos和SDC1-low-Exos，它们分别负载有上调的syndecan-1和下调的syndecan-1。与SDC1-low-Exos相比，给予SDC1-high-Exos可以改善肺水肿和炎症，减少支气管肺泡灌洗液（BALF）中的细胞数量和蛋白质水平，并保留糖萼。此外，SDC1-high-Exos 还可以减轻 LPS 攻击后促炎细胞因子的表达，例如 IL-1 beta、TNF-alpha 和 IL-6。在 MPMVEC 中，SDC1-high-Exos 减少了 LPS 刺激后的应力纤维形成并改善了单层高渗透性。 Western blotting分析表明FAK/p190RhoGAP/FthoA/ROCK/NF-kappa B信号通路可能参与LPS诱导的ALL。综上所述，SDC1-high-Exos在改善LPS刺激的ALI模型中发挥着关键作用，并可能作为未来临床应用的潜在治疗剂。
Acute lung injury (ALI) is characterized by protein-rich pulmonary edema, critical hypoxemia, and influx of proinflammatory cytokines and cells. There are currently no effective pharmacon therapies in clinical practice. Syndecan-1 in endothelial cells has potential to protect barrier function of endothelium and suppress inflammation response. Thus, the present study was to identify whether exosomes with encapsulation of syndecan-1 could achieve ideal therapeutic effects in ALI. Exosomes were isolated from the conditional medium of lentivirus-transfected mouse pulmonary microvascular endothelial cells (MPMVECs) and characterized by nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), and western blotting. ALI mouse models were induced via intratracheal administration of lipopolysaccharide (LPS) and treated with exosomes. Lung edema, inflammation, and glycocalyx thickness were examined. The possible mechanism was verified by immunoblotting in MPMVECs. The purified exosomes included SDC1-high-Exos and SDC1-low-Exos which loaded with up-regulated syndecan-1 and down-regulated syndecan-1 respectively. Compared with SDC1-low-Exos, administration of SDC1-high-Exos could ameliorate lung edema and inflammation, attenuate number of cells and protein levels in bronchoalveolar lavage fluid (BALF), and preserve glycocalyx. Furthermore, SDC1-high-Exos also mitigated the expression of pro-inflammatory cytokines such as IL-1 beta, TNF-alpha, and IL-6 following LPS challenge. In MPMVECs, SDC1-high-Exos decreased stress fiber formation and ameliorated monolayer hyper-permeability after LPS stimulation. Western blotting analysis demonstrated that FAK/p190RhoGAP/FthoA/ROCK/NF-kappa B signaling pathway may be involved in LPS-induced ALL In conclusion, SDC1-high-Exos play a pivotal role in ameliorating LPS-stimulated ALI models and may be served as a potential therapeutic agent for clinical application in the future.