人脐静脉内皮细胞来源外泌体有助于促进缺血性脑卒中小鼠的康复  

HUVECs-exo intravenous injection can promote the recovery of ischemic stroke in transient ischemic mouse model of the middle cerebral artery

作　　者：张晓星 胡慧 李跃华[1] ZHANG Xiao-xing;HU Hui;LI Yue-hua(Department of Radiology,the Sixth People’s Hospital,Shanghai Jiao Tong University,Shanghai 200233,China)

机构地区：[1]上海交通大学附属第六人民医院放射科

出　　处：《中国临床医学》2019年第3期425-431,共7页Chinese Journal of Clinical Medicine

基　　金：国家自然科学基金(81871329,81671673);上海申康新兴前沿技术联合攻关项目(SHDC12018117);上海申康医院发展中心临床科技创新项目(SHDC22015038);上海市科委医学引导项目(16411968900);上海市卫计委优秀学科带头人(2017BR041);上海市重中之重医学影像重点学科(2017ZZ02005);上海交通大学医学院高峰高原计划-“研究型医师”(2016427)~~

摘　　要：目的:探讨人脐静脉内皮细胞(HUVECs)来源的外泌体对缺血性脑卒中小鼠的治疗效果。方法:提取并鉴定HUVECs细胞来源的外泌体(HUVECs-exo),尾静脉注射30μg(溶于100μLPBS中)外泌体至短暂性大脑中动脉缺血性模型小鼠体内,对照组注射同等体积的PBS。术后第1、3、7、14、21、28天对小鼠进行mNSS评分,磁共振成像检测小鼠的梗死范围,在第28天将小鼠断头取脑,CD31/BrdU、DCX/BrdU、NeuN/BrdU免疫荧光双染评价小鼠梗死边缘区血管及神经元的新生,Western印迹法检测小鼠梗死边缘区突触前膜蛋白Synaptophysin及突触后膜蛋白PSD-95的表达情况,进而检测突触重塑。结果:HUVECs-exo组小鼠梗死范围显著小于PBS对照组(P<0.05),梗死边缘区新生血管、神经前体细胞及成熟神经元显著高于PBS对照组(P<0.05),HUVECs-exo组突触前膜蛋白Synaptophysin及后膜蛋白PSD-95表达含量显著高于PBS对照组(P<0.05)。HUVECs-exo组及PBS对照组在造模后第1、3、7天,神经功能评分无明显差异,在第14、21、28天HUVECs-exo组神经功能评分显著低于PBS对照组(P<0.05)。结论:HUVECs-exo尾静脉注射能够减小缺血性脑卒中后的梗死范围,促进梗死边缘区血管及神经元新生,促进突触重塑,改善卒中后的神经功能预后。Objective: To investigate the therapeutic effect of exosomes drived from human umbilical vein endothelial cells (HUVECs) on ischemic stroke. Methods: Exosomes derived from HUVECs were isolated and identified. Exosomes of 30 μg (dissolved in 100 μL of PBS) or an equal volume of PBS were injected intravenously into the transient ischemic mouse model of the middle cerebral artery. The mNSS scores of mice were evaluated after ischemic stroke on pos-operative day 1, 3, 7, 14, and 28, respectively. MRI imaging was used to observe the infract size. All mice were sacrificed on day 28 after surgery, CD31/ BrdU, DCX/BrdU, and NeuN/BrdU immunofluorescence double staining were applied to detected the angiogenesis and neurogenesis in the ischemic boundary zone. In order to evaluate the synaptic plasticity, the expressions of synaptophysin and PSD-95 in the ischemic boundary zone were detected using Western blotting. Results: The infarct size of the HUVECs-exo group was significantly reduced compared to the PBS group ( P <0.05). Compared to the PBS group, newborn blood vessels, neural precursor cells, and mature neurons in the ischemic boundary were significantly higher in the HUVECs-exo group, respectively ( P <0.05). The expressions of synaptophysin and PSD-95 in the ischemic boundary of the HUVECs-exo group were significantly higher compared to the PBS group ( P <0.05). There was no significant difference in mNSS scores of mice between the HUVECs-group and PBS group on day 1, 3, and 7 after surgery, but mNSS scores were significantly lower in the HUVECs-exo group on day 14, 21, and 28 compared to the PBS group ( P <0.05). Conclusions: HUVECs-exo can reduce infract size, improve angiogenesis, neurogenesis, and synaptic plasticity in the ischemic boundary zone after ischemic stroke and finally improve the neurological outcome in transient ischemic mouse model.

关 键 词：缺血性脑卒中 外泌体 神经新生 血管新生 突触重塑 

分 类 号：R743.3[医药卫生—神经病学与精神病学] R332[医药卫生—临床医学]