急性呼吸窘迫综合征（ARDS）并发脓毒症综合征（SS）的治疗仍然具有挑战性。 探讨脂肪源性间充质干细胞（ADMSCs）衍生的外泌体（EXAD）和外源性线粒体（mitoEx）相结合是否可以保护肺部来自 ARDS 并发 SS。体外研究，包括用脂多糖 (LPS) 处理的 L2 细胞，以及体内研究，包括分为组 1（假手术对照）、2（ARDS-SS）、3 组的雄性成年 SD 大鼠(ARDS-SS EXAD)、4 (ARDS-SS mitoEx) 和 5 (ARDS-SS EXAD mitoEx) 均包含在本研究中。体外研究表明，在受体 L2 细胞中发现了丰富的 mitoEx，从而显着较高的线粒体细胞色素 C、三磷酸腺苷和相对线粒体 DNA 水平 (P < 0.001)。炎症蛋白水平[白细胞介素（IL）-1β/肿瘤坏死因子（TNF）-α/核因子-κB/Toll样受体（TLR）-4/基质金属蛋白酶（MMP）-9/氧化应激（在经过 EXAD 处理的脂多糖 (LPS) 处理的 L2 细胞中，与未经 EXAD 处理相比，NOX-1/NOX-2)/细胞凋亡 (cleaved-caspase3/cleaved-poly (ADP-ribose)polymerase)] 显着减弱，而蛋白质表达细胞连接[闭塞/β-连环蛋白/闭塞小带 (ZO)-1/E-钙粘蛋白]表现出相反的炎症模式（所有 P < 0.001）。 48小时-ARDS诱导后72小时处死动物，并收获肺组织。 72小时时，支气管肺泡灌洗液的流式细胞术分析表明，炎症细胞（Ly6G / CD14 / CD68 / CD11b / c /髓过氧化物酶）和白蛋白的水平在第1组中最低，在第2组中最高，在第3组中显着更高和 4 比第 5 组高（所有 P < 0.0001），而动脉血氧饱和度 (SaO2%) 在各组中显示出相反的白蛋白模式。肺损伤/纤维化区域和炎症/DNA 损伤标记物 (CD68 /γ-H2AX) 的组织病理学结果显示各组间 SaO2% 的模式相同（所有 P < 0.0001）。炎症蛋白(TLR-4/MMP-9/IL-1β/TNF-α)/氧化应激(NOX-1/NOX-2/p22phox/氧化蛋白)/线粒体损伤(胞质-细胞色素-C/动力相关蛋白 1)/自噬（beclin-1/Atg-5/LC3B-II/LC3B-I 比率）生物标志物表现出类似的方式，而抗氧化剂 [核呼吸因子 (Nrf)-1/Nrf-2]/细胞连接（ZO-1/E-钙粘蛋白）/线粒体电子传递链（复合物 I-V）在各组中表现出相反的白蛋白方式（所有 P < 0.0001）。 EXAD-mitoEx 组合疗法比单独一种疗法更能保护肺部对抗ARDS-SS引起的伤害。©作者2024。百事登出版集团有限公司出版。保留所有权利。

The treatment of acute respiratory distress syndrome (ARDS) complicated by sepsis syndrome (SS) remains challenging.To investigate whether combined adipose-derived mesenchymal-stem-cells (ADMSCs)-derived exosome (EXAD) and exogenous mitochondria (mitoEx) protect the lung from ARDS complicated by SS.In vitro study, including L2 cells treated with lipopolysaccharide (LPS) and in vivo study including male-adult-SD rats categorized into groups 1 (sham-operated-control), 2 (ARDS-SS), 3 (ARDS-SS + EXAD), 4 (ARDS-SS + mitoEx), and 5 (ARDS-SS + EXAD + mitoEx), were included in the present study.In vitro study showed an abundance of mitoEx found in recipient-L2 cells, resulting in significantly higher mitochondrial-cytochrome-C, adenosine triphosphate and relative mitochondrial DNA levels (P < 0.001). The protein levels of inflammation [interleukin (IL)-1β/tumor necrosis factor (TNF)-α/nuclear factor-κB/toll-like receptor (TLR)-4/matrix-metalloproteinase (MMP)-9/oxidative-stress (NOX-1/NOX-2)/apoptosis (cleaved-caspase3/cleaved-poly (ADP-ribose) polymerase)] were significantly attenuated in lipopolysaccharide (LPS)-treated L2 cells with EXAD treatment than without EXAD treatment, whereas the protein expressions of cellular junctions [occluding/β-catenin/zonula occludens (ZO)-1/E-cadherin] exhibited an opposite pattern of inflammation (all P < 0.001). Animals were euthanized by 72 h post-48 h-ARDS induction, and lung tissues were harvested. By 72 h, flow cytometric analysis of bronchoalveolar lavage fluid demonstrated that the levels of inflammatory cells (Ly6G+/CD14+/CD68+/CD11b/c+/myeloperoxidase+) and albumin were lowest in group 1, highest in group 2, and significantly higher in groups 3 and 4 than in group 5 (all P < 0.0001), whereas arterial oxygen-saturation (SaO2%) displayed an opposite pattern of albumin among the groups. Histopathological findings of lung injury/fibrosis area and inflammatory/DNA-damaged markers (CD68+/γ-H2AX) displayed an identical pattern of SaO2% among the groups (all P < 0.0001). The protein expressions of inflammatory (TLR-4/MMP-9/IL-1β/TNF-α)/oxidative stress (NOX-1/NOX-2/p22phox/oxidized protein)/mitochondrial-damaged (cytosolic-cytochrome-C/dynamin-related protein 1)/autophagic (beclin-1/Atg-5/ratio of LC3B-II/LC3B-I) biomarkers exhibited a similar manner, whereas antioxidants [nuclear respiratory factor (Nrf)-1/Nrf-2]/cellular junctions (ZO-1/E-cadherin)/mitochondrial electron transport chain (complex I-V) exhibited an opposite manner of albumin among the groups (all P < 0.0001).Combined EXAD-mitoEx therapy was better than merely one for protecting the lung against ARDS-SS induced injury.©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.