Asadzadeh Bavil Mahdiyeh, Farjah Gollam Hossein, Pourheydar Bagher, Rahbarghazi Reza
Neurophysiology Research Center, Department of Anatomy, Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran.
Stem Cell Research Center, Tabriz University of University of Medical Sciences, Tabriz, Iran.
Adv Pharm Bull. 2025 May 30;15(2):406-415. doi: 10.34172/apb.025.43118. eCollection 2025 Jul.
Spinal cord ischemia-reperfusion injury (SCII) is initiated following the occlusion of supporting blood vessels, leading to the loss of neurological function. Here, we aimed to study the regenerative properties of tourniquet-induced hindlimb ischemia exosomes (Exos) in SCII Wistar rats.
Exos were isolated from rats following tourniquet-induced hindlimb ischemia. CellTracker CM-DiI-labeled Exos were injected systematically into SCII rats subjected to 60 min of abdominal aorta occlusion. The distribution of Exos was monitored using an immunofluorescence assay. Using histological examination and real-time PCR analysis, glial cell number, pyknotic and swollen neurons, and expression of apoptosis genes were studied. Oxidative stress was examined by measuring the SOD, GPx activity, MDA, and TAC levels. The neurological assessments were also performed 72 hours after the Exo injection.
Data revealed cup-shaped spherical Exos with average size and zeta potential of 279.3 nm and 15.6 mV, respectively. The isolated particles were CD9+, CD63+, and CD81+, indicating the existence of typical Exo biomarkers. Histological analysis showed reduced gliosis, pyknotic, and swollen neurons compared to SCII rats after Exos injection (<0.05). Data indicated the existence of Exos at the site of injury 24 hours after systemic injection. The injection of hypoxic Exos led to inhibition of apoptosis [Bax (0.6-fold↓), and Bcl-2 (3.97-fold↑)] and reduction of oxidative stress [MDA (58%↓), SOD (310%↑), GPx (260%↑), and TAC (300%↑)] compared to SCII rats (<0.05). Neurological assessments revealed the reduction of withdrawal response and motor deficit index in SCII rats after injection of hypoxic Exos.
Hypoxic Exos are valid regenerative tools for the alleviation of spinal cord injury (SCI) following ischemic/reperfusion.
脊髓缺血再灌注损伤（SCII）是在支持血管闭塞后引发的，会导致神经功能丧失。在此，我们旨在研究止血带诱导的后肢缺血外泌体（Exos）对SCII Wistar大鼠的再生特性。
从止血带诱导后肢缺血的大鼠中分离外泌体。将细胞追踪器CM-DiI标记的外泌体经系统注射到经历60分钟腹主动脉闭塞的SCII大鼠体内。使用免疫荧光测定法监测外泌体的分布。通过组织学检查和实时PCR分析，研究胶质细胞数量、固缩和肿胀神经元以及凋亡基因的表达。通过测量超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GPx）活性、丙二醛（MDA）和总抗氧化能力（TAC）水平来检测氧化应激。在外泌体注射72小时后也进行了神经学评估。
数据显示呈杯状的球形外泌体，平均大小和zeta电位分别为279.3 nm和15.6 mV。分离出的颗粒CD9 +、CD63 +和CD81 +，表明存在典型的外泌体生物标志物。组织学分析显示，与注射外泌体后的SCII大鼠相比，胶质增生、固缩和肿胀神经元减少（<0.05）。数据表明全身注射24小时后损伤部位存在外泌体。与SCII大鼠相比，注射低氧外泌体导致细胞凋亡受到抑制[Bax（0.6倍↓）和Bcl-2（3.97倍↑）]，氧化应激降低[MDA（58%↓）、SOD（310%↑）、GPx（260%↑）和TAC（300%↑）]（<0.05）。神经学评估显示，注射低氧外泌体后，SCII大鼠的退缩反应和运动缺陷指数降低。
低氧外泌体是减轻缺血/再灌注后脊髓损伤（SCI）的有效再生工具。
