间充质干细胞衍生的细胞外囊泡可减轻失血性休克和创伤引起的肺血管通透性和肺损伤。
Mesenchymal stem cell-derived extracellular vesicles attenuate pulmonary vascular permeability and lung injury induced by hemorrhagic shock and trauma.
机构信息
From The Department of Laboratory Medicine (D.R.P., B.Y.M., S.L.G., A.T., S.P.), University of California at San Francisco; Blood Systems Research Institute (X.D., P.P.T., S.P.), San Francisco, California; Department of Pediatric Surgery (A.K.S.), McGovern Medical School, University of Texas Health Sciences Center at Houston, Houston, Texas; Cardiovascular Research Institute (R.H.C.), University of California at San Francisco, San Francisco, California; Department of Surgery (J.B.H.), University of Texas, Houston, Texas; and Department of Surgery (M.A.S.), Oregon Health & Science University, Portland, Oregon.
出版信息
J Trauma Acute Care Surg. 2018 Feb;84(2):245-256. doi: 10.1097/TA.0000000000001744.
BACKGROUND
Mesenchymal stem cells (MSCs) have been shown to mitigate vascular permeability in hemorrhagic shock (HS) and trauma-induced brain and lung injury. Mechanistically, paracrine factors secreted from MSCs have been identified that can recapitulate many of the potent biologic effects of MSCs in animal models of disease. Interestingly, MSC-derived extracellular vesicles (EVs), contain many of these key soluble factors, and have therapeutic potential independent of the parent cells. In this study we sought to determine whether MSC-derived EVs (MSC EVs) could recapitulate the beneficial therapeutic effects of MSCs on lung vascular permeability induced by HS in mice.
METHODS
Mesenchymal stem cell EVs were isolated from human bone marrow-derived MSCs by ultracentrifugation. A mouse model of fixed pressure HS was used to study the effects of shock, shock + MSCs and shock + MSC EVs on lung vascular endothelial permeability. Mice were administered MSCs, MSC EVs, or saline IV. Lung tissue was harvested and assayed for permeability, RhoA/Rac1 activation, and for differential phosphoprotein expression. In vitro, human lung microvascular cells junctional integrity was evaluated by immunocytochemistry and endothelial cell impedance assays.
RESULTS
Hemorrhagic shock-induced lung vascular permeability was significantly decreased by both MSC and MSC EV infusion. Phosphoprotein profiling of lung tissue revealed differential activation of proteins and pathways related to cytoskeletal rearrangement and regulation of vascular permeability by MSCs and MSC EVs. Lung tissue from treatment groups demonstrated decreased activation of the cytoskeletal GTPase RhoA. In vitro, human lung microvascular cells, MSC CM but not MSC-EVs prevented thrombin-induced endothelial cell permeability as measured by electrical cell-substrate impedance sensing system and immunocytochemistry of VE-cadherin and actin.
CONCLUSION
Mesenchymal stem cells and MSC EVs modulate cytoskeletal signaling and attenuate lung vascular permeability after HS. Mesenchymal stem cell EVs may potentially be used as a novel "stem cell free" therapeutic to treat HS-induced lung injury.
背景
间充质干细胞（MSCs）已被证明可减轻失血性休克（HS）和创伤引起的脑和肺损伤中的血管通透性。从机制上讲，已经确定了 MSC 分泌的旁分泌因子，这些因子可以在疾病动物模型中再现 MSC 的许多有效生物学作用。有趣的是，MSC 衍生的细胞外囊泡（EVs）包含许多关键的可溶性因子，并且具有独立于亲本细胞的治疗潜力。在这项研究中，我们试图确定 MSC 衍生的 EV（MSC EVs）是否可以再现 MSCs 对 HS 诱导的小鼠肺血管通透性的有益治疗作用。
方法
通过超速离心从人骨髓来源的 MSC 中分离 MSC EV。使用固定压力 HS 小鼠模型研究休克、休克+MSCs 和休克+MSC EV 对肺血管内皮通透性的影响。小鼠静脉注射 MSCs、MSC EVs 或生理盐水。收获肺组织并检测通透性、RhoA/Rac1 激活和差异磷酸蛋白表达。在体外，通过免疫细胞化学和内皮细胞阻抗测定评估人肺微血管细胞连接完整性。
结果
MSCs 和 MSC EV 输注均可显著降低 HS 诱导的肺血管通透性。肺组织的磷酸蛋白谱分析显示，MSC 和 MSC EV 与细胞骨架重排和血管通透性调节相关的蛋白和途径的差异激活。治疗组的肺组织显示细胞骨架 GTPase RhoA 的激活减少。在体外，人肺微血管细胞，MSC CM 但不是 MSC-EVs 可防止凝血酶诱导的内皮细胞通透性，如通过电细胞-底物阻抗传感系统和 VE-cadherin 和肌动蛋白的免疫细胞化学测量。
结论
MSCs 和 MSC EV 调节细胞骨架信号并减轻 HS 后肺血管通透性。MSC EV 可能可作为一种新型的“无干细胞”治疗方法，用于治疗 HS 诱导的肺损伤。
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