Meng Qinghe, Winston Tackla, Ma Julia, Song Yuanhui, Wang Chunyan, Yang Junhui, Ma Zhen, Cooney Robert N
Department of Surgery, State University of New York (SUNY), Upstate Medical University, Syracuse, New York.
Department of Biomedical & Chemical Engineering, Syracuse University, Syracuse, New York.
Shock. 2024 Aug 1;62(2):294-303. doi: 10.1097/SHK.0000000000002381. Epub 2024 May 23.
Introduction: We hypothesized extracellular vesicles (EVs) from preconditioned human-induced pluripotent stem cell-derived mesenchymal stem cells (iMSCs) attenuate LPS-induced acute lung injury (ALI) and endotoxemia. Methods: iMSCs were incubated with cell stimulation cocktail (CSC) and EVs were isolated. iMSC-EVs were characterized by size and EV markers. Biodistribution of intratracheal (IT), intravenous, and intraperitoneal injection of iMSC-EVs in mice was examined using IVIS. Uptake of iMSC-EVs in lung tissue, alveolar macrophages, and RAW264.7 cells was also assessed. C57BL/6 mice were treated with IT/IP iMSC-EVs or vehicle ± IT/IP LPS to induce ALI/acute respiratory distress syndrome and endotoxemia. Lung tissues, plasma, and bronchoalveolar lavage fluid (BALF) were harvested at 24 h. Lung histology, BALF neutrophil/macrophage, cytokine levels, and total protein concentration were measured to assess ALI and inflammation. Survival studies were performed using IP LPS in mice for 3 days. Results: iMSC-EV route of administration resulted in differential tissue distribution. iMSC-EVs were taken up by alveolar macrophages in mouse lung and cultured RAW264.7 cells. IT LPS-treated mice demonstrated marked histologic ALI, increased BALF neutrophils/macrophages and protein, and increased BALF and plasma TNF-α/IL-6 levels. These parameters were attenuated by 2 h before or 2 h after treatment with IT iMSC-EVs in ALI mice. Interestingly, the IT LPS-induced increase in IL-10 was augmented by iMSC-EVs. Mice treated with IP LPS showed increases in TNF-α and IL-6 that were downregulated by iMSC-EVs and LPS-induced mortality was ameliorated by iMSC-EVs. Administration of IT iMSC-EVs 2 h after LPS downregulated the increase in proinflammatory cytokines (TNF-α/IL-6) by LPS and further increased IL-10 levels. Conclusions: iMSC-EVs attenuate the inflammatory effects of LPS on cytokine levels in ALI and IP LPS in mice. LPS-induced mortality was improved with administration of iMSC-EVs.
我们假设经预处理的人诱导多能干细胞来源的间充质干细胞（iMSC）分泌的细胞外囊泡（EV）可减轻脂多糖（LPS）诱导的急性肺损伤（ALI）和内毒素血症。方法：将iMSC与细胞刺激鸡尾酒（CSC）孵育并分离出EV。通过大小和EV标志物对iMSC-EV进行表征。使用IVIS检测小鼠气管内（IT）、静脉内和腹腔内注射iMSC-EV后的生物分布。还评估了iMSC-EV在肺组织、肺泡巨噬细胞和RAW264.7细胞中的摄取情况。用IT/IP iMSC-EV或赋形剂±IT/IP LPS处理C57BL/6小鼠以诱导ALI/急性呼吸窘迫综合征和内毒素血症。在24小时时收集肺组织、血浆和支气管肺泡灌洗液（BALF）。测量肺组织学、BALF中性粒细胞/巨噬细胞、细胞因子水平和总蛋白浓度以评估ALI和炎症。使用IP LPS对小鼠进行3天的生存研究。结果：iMSC-EV的给药途径导致不同的组织分布。iMSC-EV被小鼠肺中的肺泡巨噬细胞和培养的RAW264.7细胞摄取。IT LPS处理的小鼠表现出明显的组织学ALI、BALF中性粒细胞/巨噬细胞和蛋白质增加，以及BALF和血浆TNF-α/IL-6水平升高。在ALI小鼠中，在IT iMSC-EV处理前2小时或处理后2小时，这些参数均有所减轻。有趣的是，iMSC-EV增强了IT LPS诱导的IL-10增加。用IP LPS处理的小鼠显示TNF-α和IL-6增加，iMSC-EV使其下调，并且iMSC-EV改善了LPS诱导的死亡率。在LPS后2小时给予IT iMSC-EV可下调LPS诱导的促炎细胞因子（TNF-α/IL-6）增加，并进一步提高IL-10水平。结论：iMSC-EV减轻了LPS对小鼠ALI和IP LPS中细胞因子水平的炎症作用。给予iMSC-EV可改善LPS诱导的死亡率。
