Xu Minghua, Zhao Xiaomin, Zhao Jiang, Tan Zhisheng, Zhang Chengshi, Huang Yun, Zhong Huiping, Guo Meifeng, Zhang Chen, Ye Ping, Zheng Wentao
Intensive-Care Unit Punan Branch of Renji Hospital Shanghai Jiao Tong University School of Medicine, Shanghai 200125, China.
Department of Neurosurgery Punan Branch of Renji Hospital Shanghai Jiao Tong University School of Medicine, Shanghai 200125, China.
Appl Bionics Biomech. 2024 Jul 25;2024:2550642. doi: 10.1155/2024/2550642. eCollection 2024.
Moyamoya disease (MMD) leads to nerve injury. Exosomes are touted as bio-shuttles for the delivery of distinct biomolecules inside the cells. Recently, UCH-L1 was shown to play a vital role in nerve injury. However, it is still unknown whether UCH-L1 can improve the nerve injury of MMD.
Exosomes were isolated from the serum of patients with MMD and healthy controls. The total RNA was extracted from the exosomes, and the level of GFAP and UCH-L1 between the serum exosomes of the two groups was analyzed by a quantitative reverse transcription-polymerase chain reaction and western blot. Exosome labeling and uptake by SH-SY5Y cells were observed by confocal laser microscopy. Cell counting kit-8 assay and flow cytometry were used to determine the viability and apoptosis of SH-SY5Y cells, respectively.
Exosomes were successfully isolated and identified from serum. The expression of GFAP and UCH-L1 was significantly higher in the serum-derived exosomes from MMD patients compared with the healthy controls ( < 0.05). Compared to the blank and control exosome group, serum-derived exosomes from MMD significantly suppress cellular vitality and promote apoptosis of SH-SY5Y cells, while the use of LDN-91946, a specific inhibitor of UCH-L1, could reverse the effects induced by serum-derived exosomes from MMD.
UCH-L1 inhibitor could reverse MMD-induced inhibition of SH-SY5Y cell viability and promotion of apoptosis. UCH-L1 may be a therapeutic target for the treatment of nerve damage caused by MMD.
烟雾病（MMD）会导致神经损伤。外泌体被视为在细胞内递送不同生物分子的生物载体。最近，泛素羧基末端水解酶L1（UCH-L1）被证明在神经损伤中起关键作用。然而，UCH-L1是否能改善烟雾病的神经损伤仍不清楚。
从烟雾病患者和健康对照者的血清中分离外泌体。从外泌体中提取总RNA，通过定量逆转录聚合酶链反应和蛋白质免疫印迹法分析两组血清外泌体中神经胶质纤维酸性蛋白（GFAP）和UCH-L1的水平。通过共聚焦激光显微镜观察SH-SY5Y细胞对外泌体的标记和摄取。分别使用细胞计数试剂盒-8法和流式细胞术测定SH-SY5Y细胞的活力和凋亡情况。
成功从血清中分离并鉴定出外泌体。与健康对照相比，烟雾病患者血清来源的外泌体中GFAP和UCH-L1的表达显著更高（<0.05）。与空白和对照外泌体组相比，烟雾病患者血清来源的外泌体显著抑制SH-SY5Y细胞的活力并促进其凋亡，而使用UCH-L1的特异性抑制剂LDN-91946可逆转烟雾病患者血清来源外泌体诱导的效应。
UCH-L1抑制剂可逆转烟雾病诱导的对SH-SY5Y细胞活力的抑制和凋亡的促进。UCH-L1可能是治疗烟雾病所致神经损伤的一个治疗靶点。
