检索历史 清除

目的:探讨远端缺血适应来源的外泌体对大鼠心脏骤停后神经功能障碍的治疗作用。方法:Sprague-Dawley大鼠在经历远端缺血适应后，从血浆中分离出外泌体用作干预药物。36只大鼠被随机分为假手术组、对照组和外泌体组（ n=12）。对照组和外泌体组大鼠使用窒息法诱导心脏骤停，恢复自主循环后分别静脉输注安慰剂或外泌体悬液（1×10 10 Particles）。神经功能缺陷评分、旷场试验、Y迷宫和水迷宫用以评估神经功能结果。检测血浆透明质酸（hyaluronic acid, HA）和多配体蛋白聚糖-1（syndecan-1, Sdc-1）水平、检测海马CA1区基质金属蛋白酶2/9（matrix metalloproteinase-2/9，MMP-2/9）的表达水平。 结果:在经历远端缺血适应后，大鼠血浆中外泌体含量较正常大鼠升高。和对照组相比，外泌体组大鼠行为学实验明显改善，表现为水平运动距离增加（5.86±2.89 vs. 17.53±5.51， P<0.05）、自发交替正确率升高（13.29±15.07 vs. 42.63±10.25， P<0.05）、逃避潜伏期缩短（25.83±8.54 vs. 13.49±4.55， P<0.05）。复苏后24 h的血浆HA和Sdc-1水平明显降低（HA：26.34±9.83 vs. 14.84±6.26， P<0.05；Sdc-1：0.05±0.03 vs. 0.02±0.02， P<0.05），同时海马组织中MMP-2/9水平明显降低。 结论:从经历远端缺血适应的大鼠血浆中提取出的外泌体能够改善大鼠心脏骤停后神经功能障碍，其机制可能与MMP-2/9的减少以内皮糖萼降解减少有关。

Objective:To investigate the therapeutic effect of exosomes derived from remote ischemic conditioning on neurological dysfunction after cardiopulmonary resuscitation in a rat model of cardiac arrest and the relationship with glycocalyx protection.Methods:Exosomes were isolated from the blood of healthy adult male Sprague-Dawley rats using ultracentrifugation after undergoing remote ischemic conditioning for use as intervention drugs. Nanoparticle tracking analysis technology was used for exosome detection. Thirty-six adult male Sprague-Dawley rats were randomly assigned to 3 groups ( n=12 each) :Sham group, Control group and Exosome group. Cardiac arrest was induced by asphyxia for 7 min in the Control and Exosome groups. Placebo or exosomes (1×10 10 Particles) were infused intravenously at 5 min after the rats had returned of spontaneous circulation. Neuropsychological deficit score (NDS), open field test, Y maze and Morris water maze were used to assess neurological outcomes. The levels of plasma Hyaluronic acid (HA) and syndecan-1 (Sdc-1) were detected by Elisa. The expression levels of matrix metalloproteinase-2/9 (MMP-2/9) in hippocampal CA1 region were detected by Western blot. Results:After undergoing remote ischemic conditioning, the plasma levels of exosomes were elevated in rats compared to normal rats. Compared with the control group, the behavioral experiment of rats in the exosomes group were significantly improved, as evidenced by an increase in horizontal locomotor distance (5.86±2.89 vs. 17.53±5.51, P< 0.05), an increase in the correct rate of spontaneous alternation (13.29±15.07 vs. 42.63±10.25, P< 0.05), and a shortening of avoidance latency (25.83±8.54 vs. 13.49±4.55, P< 0.05). Plasma HA and Sdc-1 levels were significantly lower 24 h after resuscitation (HA: 26.34±9.83 vs. 14.84±6.26, P< 0.05; Sdc-1: 0.05±0.03 vs. 0.02±0.02, P<0.05), along with significantly lower MMP-2/9 levels in hippocampal tissue. Conclusions:Exosomes extracted from the plasma of rats undergoing remote ischemic conditioning can improve neurological dysfunction after cardiac arrest in rats, and the mechanism may be related to the inhibition of metalloproteinases and the reduction of endothelial glycocalyx degradation.