中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2022, Vol. 17 ›› Issue (1): 194-202.doi: 10.4103/1673-5374.314323

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

骨髓间充质干细胞来源外泌体可抑制周细胞焦亡保护损伤的脊髓

  

  • 出版日期:2022-01-05 发布日期:2021-09-22

Exosomes derived from bone marrow mesenchymal stem cells protect the injured spinal cord by inhibiting pericyte pyroptosis

Yan Zhou1, #, Lu-Lu Wen2, #, Yan-Fei Li2, Kai-Min Wu2, Ran-Ran Duan2, Yao-Bing Yao2, Li-Jun Jing2, Zhe Gong2, Jun-Fang Teng2, Yan-Jie Jia2, *   

  1. 1Department of Radiology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, China; 2Department of Neurology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, China
  • Online:2022-01-05 Published:2021-09-22
  • Contact: Yan-Jie Jia, PhD, jiayanjie1971@edu.zzu.cn.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China, No. U1604170 (to YJJ).

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摘要：

间充质干细胞移植已被认为是治疗脊髓损伤的潜在策略，但肿瘤发生和免疫排斥等问题限制了其临床应用。由于间充质干细胞的疗效主要依赖于旁分泌作用，而外泌体是间充质干细胞释放旁分泌因子中最重要的成分，因而可能替代间充质干细胞移植，但是其潜在机制仍有待探索。试验以打击法建立T10脊髓损伤大鼠模型，在损伤后30min和1d时经尾静脉注射200μL外泌体（200μg/mL，来自约1×106 骨髓间充质干细胞）进行治疗。结果显示，（1）骨髓间充质干细胞来源的外泌体可显著减少脊髓损伤后神经元的死亡，改善髓鞘的无序排列和脱失，增加脊髓血管壁上周细胞/内皮细胞覆盖，减轻血脊髓屏障渗漏，同时减少Caspase 1的阳性表达，抑制白细胞介素1β的释放，促进大鼠运动功能的恢复；（2）在体外以干扰素γ、肿瘤坏死因子α、脂多糖、 Lipofectamine®3000以及ATP复合刺激诱导周细胞焦亡模拟体外损伤，以骨髓间充质干细胞来源的外泌体预处理8h，可明显减轻周细胞的焦亡，并提高周细胞的存活率。（3）实验结果表明，骨髓间充质干细胞来源的外泌体可通过抑制周细胞焦亡来保护血脊髓屏障的完整性，促进神经元的存活和神经纤维的延伸，最终促进脊髓损伤大鼠运动功能的恢复。实验于2019年3月16日经郑州大学伦理委员会批准。

https://orcid.org/0000-0002-2220-9257 (Yan-Jie Jia); https://orcid.org/0000-0001-9571-6295 (Yan Zhou); https://orcid.org/0000-0002-9225-5443 (Lu-Lu Wen)

关键词: 脊髓损伤, 血脊髓屏障, 血肿, 周细胞, 外泌体, 焦亡, pro-caspase 1, NOD1

Abstract: Mesenchymal stem cell (MSC) transplantation is a promising treatment strategy for spinal cord injury, but immunological rejection and possible tumor formation limit its application. The therapeutic effects of MSCs mainly depend on their release of soluble paracrine factors. Exosomes are essential for the secretion of these paracrine effectors. Bone marrow mesenchymal stem cell-derived exosomes (BMSC-EXOs) can be substituted for BMSCs in cell transplantation. However, the underlying mechanisms remain unclear. In this study, a rat model of T10 spinal cord injury was established using the impact method. Then, 30 minutes and 1 day after spinal cord injury, the rats were administered 200 μL exosomes via the tail vein (200 μg/mL; approximately 1 × 106 BMSCs). Treatment with BMSC-EXOs greatly reduced neuronal cell death, improved myelin arrangement and reduced myelin loss, increased pericyte/endothelial cell coverage on the vascular wall, decreased blood-spinal cord barrier leakage, reduced caspase 1 expression, inhibited interleukin-1β release, and accelerated locomotor functional recovery in rats with spinal cord injury. In the cell culture experiment, pericytes were treated with interferon-γ and tumor necrosis factor-α. Then, Lipofectamine 3000 was used to deliver lipopolysaccharide into the cells, and the cells were co-incubated with adenosine triphosphate to simulate injury in vitro. Pre-treatment with BMSC-EXOs for 8 hours greatly reduced pericyte pyroptosis and increased pericyte survival rate. These findings suggest that BMSC-EXOs may protect pericytes by inhibiting pyroptosis and by improving blood-spinal cord barrier integrity, thereby promoting the survival of neurons and the extension of nerve fibers, and ultimately improving motor function in rats with spinal cord injury. All protocols were conducted with the approval of the Animal Ethics Committee of Zhengzhou University on March 16, 2019.

Key words: blood-spinal cord barrier, edema, exosome, pericyte, NOD1, pro-caspase 1, pyroptosis, spinal cord injury