骨髓间充质干细胞来源的外泌体静脉移植对脊髓损伤的修复作用研究  

Study on the Repair Effect of Bone Marrow Mesenchymal Stem Cell-derived Exocrine Vein Transplantation on Spinal Cord Injury

作　　者：马骢 李刚 马亮亮 李想 张永泼 Ma Cong;Li Gang;Ma Liangliang(Department of Rehabilitation Medicine,Luoyang Orthopedic Hospital(Henan Orthopedic Hospital),Zhengzhou,Henan,450003,China)

机构地区：[1]河南省洛阳正骨医院(河南省骨科医院)康复医学科,河南郑州450003 [2]河南省洛阳正骨医院(河南省骨科医院)小儿外科,河南郑州450003 [3]河南省组织细胞库有限公司,河南郑州450000

出　　处：《黑龙江医学》2024年第7期787-790,共4页Heilongjiang Medical Journal

摘　　要：目的:分析骨髓间充质干细胞来源的外泌体静脉移植对脊髓损伤的修复作用。方法:选择2013年10月—2014年3月河南省洛阳正骨医院实验室试验的78例大鼠为研究对象,应用随机抛球法将大鼠分为手术组(n=26)、对照组(n=26)、外泌体组(n=26)。通过脊髓法建立大鼠脊髓损伤模型后,对照组不作处理,手术组在对照组的基础上实施切开减压术,而外泌体组在对照组的基础上实施全骨髓培养法培养大鼠BMSCs,收集P2代细胞上清,应用ExoQuickPrecipitation提取法分离并纯化外泌体,通过透射电镜观察鉴定外泌体形态,采用Westernblot鉴定外泌体表面标志蛋白CD9、CD63,在造模1 h后尾静脉给予外泌体移植500μL。分别采用BBB评分、斜板实验在术后1、3、7、14、21、28 d评价大鼠的运动功能恢复情况,并于术后28 d处死试验大鼠,采用苏木精-伊红(HE)染色、髓鞘(LFB)染色观察各组脊髓组织形态学改变,尼氏(Nissl)染色观察神经元存活数目。结果:术后各时间点手术组大鼠BBB评分高于对照组和外泌体组,术后7、14、21、28 d外泌体组大鼠BBB评分高于对照组,差异有统计学意义(P<0.05)。手术组术后各时间点斜板评分高于对照组及外泌体组,差异有统计学意义(P<0.05)。造模后28 d,手术组的脊髓组织HE染色体等均正常,神经元数目减少。结论:BMSCs外泌体静脉移植对脊髓损伤有明显的改善作用,可以提高患者的运动能力,减轻脊髓损伤,加速脊髓损伤的恢复。Objective:To analyze the repair effect of bone marrow mesenchymal stem cell-derived exocrine vein transplantation on spinal cord injury.Methods:78 rats from the laboratory test from October 2013 to March 2014 were selected as the research objects.The rats were randomly divided into the operation group(n=26),the control group(n=26),and the exocrine body group(n=26)with ball throwing method.After the construction of SCI modelling with spinal cord method,the control group received no intervention,operation group received incision decompression based on control group,and exocrine body group received overall medulloculture based on control group to culture rat BMSCs.The supernatant of P2 generation cells was collected.The exocrine body was isolated and purified by ExoQuick Precision extraction method.The morphology of exocrine body was observed and identified by transmission electron microscope,and the surface marker proteins CD9 and CD63 of exocrine body were identified by Western blot.After 1 hour of model establishment,the exocrine body was transplanted into the caudal vein for 500μL.The rats were respectively constructed with BBB score and inclined plate test.The recovery of rats’motor function was evaluated 1,3,7,14,21,and 28 days after construction,and the rats were killed 28 days after operation.The rats were stained with hematoxylin-eosin(HE),the morphological changes of spinal cord tissue in each group were observed by staining the myelin sheath(LFB),and the survival number of neurons was observed by Nissl staining.Results:The BBB score of the operation group was higher than that of the control group and the exosome group at all time after surgery,and the BBB score of the exosome group was higher than that of the control group at 7,14,21 and 28 days after surgery,with statistically significant difference(P<0.05).The oblique plate score of the operation group was higher than that of the control group and exosome group,and the difference was statistically significant(P<0.05).28 days after modeling,HE chromos

关 键 词：骨髓间充质干细胞来源 外泌体静脉移植 脊髓损伤 修复作用 

分 类 号：R651.2[医药卫生—外科学]