远端缺血适应外泌体对大鼠心脏骤停后脑损伤的保护作用
Therapeutic potential of exosome derived from remote ischemic conditioning in alleviating neurologic dysfunction after cardiac arrest in a rat model
翟恒 1王吉慧 2陈习 3张扬 2胡书群 4王军 5彭易根 6赵宁军 1燕宪亮 2苏成磊 1许铁7
作者信息
- 1. 徐州医科大学附属医院急诊医学科,徐州 221006
- 2. 徐州医科大学附属医院急诊医学科,徐州 221006;睢宁县人民医院急诊医学科,睢宁 221200
- 3. 睢宁县人民医院急诊医学科,睢宁 221200
- 4. 徐州医科大学附属医院急诊医学科,徐州 221006;徐州医科大学第二临床医学院急诊医学实验室,徐州 221006
- 5. 南京大学医学院附属鼓楼医院急诊医学科,南京 210008
- 6. 南京市江宁医院急诊医学科,南京 211100
- 7. 徐州医科大学附属医院急诊医学科,徐州 221006;南京市江宁医院急诊医学科,南京 211100;徐州医科大学江苏省卫生应急研究所,徐州 221006
- 折叠
摘要
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(1×1010 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24h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.
Abstract
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 × 1010 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.
关键词
远端缺血适应/外泌体/心脏骤停/神经保护/糖萼/透明质酸/多配体蛋白聚糖-1/基质金属蛋白酶2/基质金属蛋白酶9Key words
Remote ischemic conditioning/Exosomes/Cardiac arrest/Neuroprotection/Glycocalyx/Hyaluronic acid/Syndecan-1/Matrix metalloproteinase 2/Matrix metalloproteinase 9引用本文复制引用
基金项目
国家自然科学基金(82202426)
江苏省自然科学基金(BK20220674)
徐州市青年医学科技创新项目(XWKYHT20210571)
睿E(睿意)急诊医学研究专项基金(R2021003)
出版年
2024
NETL
NSTL
万方数据