Department of Orthopedics, The Second Affiliated Hospital of Xi'an Medical University, China.
Traditional Chinese Medicine Department, The Second Affiliated Hospital of Xi'an Medical University, China.
J Musculoskelet Neuronal Interact. 2022 Sep 1;22(3):411-421.
To evaluate the roles of MicroRNAs (miRNAs) enclosed in the neuron-derived exosomes in the recovery of the spinal cord injury (SCI) and the mechanism.
The exosomes were isolated from neural stem cells (NSCs) and characterized by transmission electron microscopy (TEM) and NanoSight system (NTA). For experiments, Basso Mouse Scale, beam walking, and inclined plane tests were used to determine the behavioral symptoms of the SCI mice. For experiments, HO treated HT22 cells were used to simulate SCI cells and cocultured with exosomes to analyze the cell apoptosis using TUNEL assays and flow cytometry. Apoptosis- and autophagy-related protein expression was detected by western blot and the green fluorescent protein (GFP)-LC3 assay was used to detect the level of autophagy. In addition, luciferase assay was performed to assess the relationship between miR-374-5p and SKT-4.
Exosomes from NSCs alleviated spinal cord injury by triggering autophagy flux and suppressing apoptosis. Besides, miR-374-5p was highly expressed in these exosomes and was responsible for the decent in injured neural cell apoptosis by activating autophagy flux. The SKT-4 was the target gene regulated by miR-374-5p in this exosomal protective function to SCI cells.
The elevated level of miR-374-5p in neuronal exosomes could enhance spinal cord injury recovery by activating autophagy.
评估神经元衍生的外泌体中包裹的 MicroRNAs(miRNAs)在脊髓损伤(SCI)恢复中的作用及机制。
通过透射电子显微镜(TEM)和纳米粒子跟踪分析(NTA)系统分离神经干细胞(NSCs)来源的外泌体并进行鉴定。在实验中,使用 Basso Mouse Scale、束走和斜面试验来确定 SCI 小鼠的行为症状。在实验中,使用 HO 处理的 HT22 细胞模拟 SCI 细胞,并与外泌体共培养,通过 TUNEL 检测和流式细胞术分析细胞凋亡。通过 Western blot 检测凋亡和自噬相关蛋白的表达,并用 GFP-LC3 检测自噬水平。此外,通过荧光素酶实验评估 miR-374-5p 与 SKT-4 之间的关系。
NSCs 的外泌体通过触发自噬流和抑制细胞凋亡来减轻脊髓损伤。此外,这些外泌体中高度表达 miR-374-5p,通过激活自噬流来降低受损神经细胞的凋亡。SKT-4 是外泌体在 SCI 细胞中发挥保护作用的 miR-374-5p 调节的靶基因。
神经元外泌体中 miR-374-5p 水平的升高可以通过激活自噬来增强脊髓损伤的恢复。