Li Lingxia, Jin Shuan, Zhang Yingmin
Department of Respiratory Diseases, First Affiliated Hospital of Henan University of Science and Technology Luoyang, China.
Department of Anesthesiology, Jinan Central Hospital, Shandong University Jinan, China.
Int J Clin Exp Med. 2015 Mar 15;8(3):3825-32. eCollection 2015.
To explore the effect of bone marrow mesenchymal stem cells (MSCs) on endotoxin-induced acute lung injury in mice and verify the role of exosome.
Exosome was isolated from the culture supernatant of MSC. For ischemic preconditioning, MSCs were subjected to anoxia for 0 min (MSCs group), 30 min (MSCs(IPC-30) group), 60 min (MSCs(IPC-60) group) and 90 min (MSCs(IPC 90) group), and then used to treat endotoxin-injured mice. The exosome from the optimal group was used to treat endotoxin-injured mice. In addition, the exosome from the optimal group was also used to treat the endotoxin-stimulated RAW 264.7 cells for 6 h and 12 h.
CD63 positive exosome were acquired through ExoQuick kits. Administration of MSCs, MSCs(IPC-30), MSCs(IPC-60) and MSCs(IPC-90) could reduce the level of white blood cells (WBC) and neutrophils into the bronchoalveolar lavage (BAL) fluid of endotoxin-injured mice, and the MSCIPC-60 group had the greatest reduction, which reduced WBC by 57% and neutrophils by 55%. Administration of MSCs(IPC-60) exosome could also reduce the level of WBC, neutrophils, MIP-2 and penetration protein into the BAL fluid of endotoxin-injured mice, which had the same effect as MSCs(IPC-60) and showed a dose dependent, compared to MSCs exosome. In addition, MSCs(IPC-60) exosome were used to treat endotoxin-stimulated RAW 264.7 cells, and the level of TNFα at 6 h and 12 h was significantly reduced, while the level of IL-10 at 12 h increased.
Ischemic preconditioning for 60 min can potentiates the protective effect of MSC on endotoxin-induced Acute Lung Injury through the secretion of Exosome.
探讨骨髓间充质干细胞（MSCs）对内毒素诱导的小鼠急性肺损伤的影响，并验证外泌体的作用。
从MSCs培养上清液中分离外泌体。对于缺血预处理，将MSCs分别进行0分钟（MSCs组）、30分钟（MSCs(IPC - 30)组）、60分钟（MSCs(IPC - 60)组）和90分钟（MSCs(IPC 90)组）的缺氧处理，然后用于治疗内毒素损伤的小鼠。将最佳组的外泌体用于治疗内毒素损伤的小鼠。此外，最佳组的外泌体还用于处理内毒素刺激的RAW 264.7细胞6小时和12小时。
通过ExoQuick试剂盒获得CD63阳性外泌体。给予MSCs、MSCs(IPC - 30)、MSCs(IPC - 60)和MSCs(IPC - 90)可降低内毒素损伤小鼠支气管肺泡灌洗（BAL）液中的白细胞（WBC）和中性粒细胞水平，其中MSCs(IPC - 60)组降低幅度最大，WBC降低了57%，中性粒细胞降低了55%。给予MSCs(IPC - 60)外泌体也可降低内毒素损伤小鼠BAL液中的WBC、中性粒细胞、MIP - 2和穿透蛋白水平，与MSCs(IPC - 60)效果相同，且与MSCs外泌体相比呈剂量依赖性。此外，用MSCs(IPC - 60)外泌体处理内毒素刺激的RAW 264.7细胞，6小时和12小时时TNFα水平显著降低，12小时时IL - 10水平升高。
60分钟的缺血预处理可通过外泌体的分泌增强MSCs对内毒素诱导的急性肺损伤的保护作用。
