骨髓间充质干细胞外泌体减轻海马神经细胞氧糖剥夺再灌注损伤  

Bone marrow mesenchymal stem cell exosomes alleviate oxygen-glucose deprivation/reperfusion injury in hippocampal neurons

作　　者：时将 高诗仑 刘金铎 谷天祥[1] 师恩祎[1] Shi Jiang;Gao Shilun;Liu Jinduo;Gu Tianxiang;Shi Enyi(Department of Cardiac Surgery,First Affiliated Hospital of China Medical University,Shenyang 110000,Liaoning Province,China)

机构地区：[1]中国医科大学附属第一医院心脏外科,辽宁省沈阳市110000

出　　处：《中国组织工程研究》2019年第21期3316-3322,共7页Chinese Journal of Tissue Engineering Research

基　　金：2016年国家自然科学基金项目(81471267);项目负责人:师恩祎~~

摘　　要：背景:深低温停循环术后中枢神经系统并发症已经日益成为人们注意的焦点,其核心机制仍是全脑缺血再灌注损伤。目的:探讨骨髓间充质干细胞外泌体对海马神经细胞氧糖剥夺再灌注损伤的保护性作用及其机制。方法:将培养至第7天的海马神经细胞随机分为正常对照组、氧糖剥夺再灌注损伤组及骨髓间充质干细胞外泌体组,后2组进行2 h氧糖剥夺再灌注损伤。造模后,3组海马神经细胞均更换为神经细胞培养基,骨髓间充质干细胞外泌体组加入骨髓间充质干细胞外泌体,正常对照组和氧糖剥夺再灌注损伤组加入等量的PBS溶液。继续培养24 h收集细胞进行Western blot检测,培养48 h进行免疫荧光染色统计海马神经细胞存活率和突起生长情况。结果与结论:①与正常对照组比较,氧糖剥夺再灌注损伤组海马神经细胞存活率明显降低,突起长度及分支数明显受损,促炎因子表达水平显著上升;②与氧糖剥夺再灌注损伤组比较,骨髓间充质干细胞外泌体组神经细胞存活率和突起长度及分支数显著增加,肝细胞生长因子和脑源性神经营养因子表达水平显著增加,促炎症因子表达水平显著下降;③结果提示,骨髓间充质干细胞外泌体能减轻海马神经细胞氧糖剥夺再灌注损伤,其机制可能与抑制促炎因子分泌和促进肝细胞生长因子、脑源性神经营养因子表达有关。BACKGROUND:Increasing attentions have been focused on postoperative neurological complications after deep hypothermic circulatory arrest.The core mechanism of neurological complications is global ischemia-reperfusion injury.OBJECTIVE:To investigate the protective effect and mechanism of bone mesenchymal stem cell exosomes on oxygen-glucose deprivation/reperfusion injury in hippocampal neurons.METHODS:Primary cultured fetal rat hippocampal neurons(7 days)were divided into control group,oxygen-glucose deprivation/reperfusion group(OGD/R group)and oxygen-glucose deprivation/reperfusion with adding mesenchymal stem cell exosomes(exosomes group).After 2 hours of oxygen-glucose deprivation/reperfusion,the hippocampal neurons in the latter two groups were cultured under normal conditions.After modeling,hippocampal neurons in all the three groups were cultured in nerve cell culture medium,followed by addition of bone marrow mesenchymal stem cell exosomes in the exosomes group,and addition of PBS in the other two groups.After 24 hours of continuous culture,western blot assay was used;after 48 hours of culture,the survival rate and the growth of neurites in hippocampal neurons were counted and observed by immunofluorescence staining.RESULTS AND CONCLUSION:Compared with the control group,the survival rate of hippocampal neurons in the OGD/R group was significantly reduced,the length of neurites and the number of branches were significantly impaired,and the expression level of pro-inflammatory factors was significantly increased.Compared with the OGD/R group,the survival rate of hippocampal neurons,the length of neuritis,and the number of branches in the exosomes group were significantly increased in the exosomes group,the expression levels of hepatocyte growth factor and brain-derived neurotrophic factor significantly increased,and the expression of pro-inflammatory factors obviously decreased.The results suggest that exosomes from bone marrow mesenchymal stem cells can alleviate oxygen-glucose deprivation/reperfusion injur

关 键 词：骨髓间充质干细胞 外泌体 海马神经细胞 氧糖剥夺再灌注损伤 深低温停循环 肝细胞生长因子 脑源性神经营养因子 环氧化酶2 诱导型一氧化氮合酶 肿瘤坏死因子α 促炎症因子 国家自然科学基金 

分 类 号：R459.9[医药卫生—治疗学] R394.[医药卫生—临床医学]