摘要
目的探讨血管生成素-1(Ang-1)基因修饰的骨髓间充质干细胞(MSCs)对脑梗死大鼠行为学的影响,为其临床应用提供依据。方法体外培养MSCs,制备转染Ang-1基因重组腺病毒的MSCs(Ang-MSCs)或空载体腺病毒的MSCs(Null-MSCs),并鉴定;制备18只大鼠大脑中动脉栓塞模型,按随机数字表法随机分为生理盐水组、Null-MSCs组和Ang-MSCs组各6只。成模后24 h,生理盐水组尾静脉注射生理盐水1 m L,Null-MSCs组尾静脉移植5.0×109/m L Null-MSCs 1 m L,Ang-MSCs组尾静脉移植5.0×109/m L Ang-MSCs 1 m L。另取大鼠6只作为假手术组,手术操作与各模型组相同,但不阻断大脑中动脉,同期尾静脉注射生理盐水1 m L。移植后1周,通过Morris水迷宫试验检测各组学习和记忆能力。结果经鉴定,Ang-MSCs可稳定表达Ang-1蛋白。移植后1周,与假手术组比较,生理盐水组、Null-MSCs组和Ang-MSCs组水迷宫寻台潜伏期明显延长,2 min内经过相应平台位置次数明显减少(P<0.05或<0.01);与生理盐水组比较,Null-MSCs组、Ang-MSCs组水迷宫寻台潜伏期明显缩短(P均<0.01),2 min内经过相应平台位置次数明显增多(P均<0.01),且以Ang-MSCs组效果更明显(P<0.05或<0.01)。结论 Ang-1基因修饰的MSCs可改善脑梗死大鼠的学习和记忆能力。
Objective To investigate the effects of adenovirus-mediated angiopoietin-1 ( Ang-1 ) gene modified bone marrow mesenchymal stem cells ( MSCs) transplantation on the behavior of the cerebral infarction rats in order to provide the basis for clinical application.Methods MSCs were isolated and cultured in vitro, and infected by the recombinant ad-enovirus vectors carrying Ang-1 gene ( Ang-MSCs cells) or the empty adenovirus ( Null-MSCs cells) .Adult male rats were subjected to transient (2 h) middle cerebral artery occlusion ( MCAO) with modified Zea Longa method.The rat models were randomly divided into four groups (n=6): sham operation group, saline group, Null-MSCs group and Ang-MSCs group.The saline, Null-MSCs and Ang-MSCs groups were respectively transplanted with 1 mL of saline, Null-MSCs cells (5.0 ×109 cell/mL) and Ang-MSCs cells (5.0 ×109 cell/mL) into the tail vein of rats at 24 h after MCAO.The middle cerebral artery was not blocked in the sham operation group, but they were trsnsplanted with 1 mL of saline into the tail vein of rats at the same time.Morris Water Maze was used to detect the spatial learning and memory abilities of rats at 1 week post-transplantation.Results Western blotting indicated that Ang-MSCs stably expressed Ang-1 protein.One week later, compared with the sham operation group, the escape latency of Water Maze was significantly lengthened and the times of crossing the original table in 2 minutes were decreased (P<0.05 or P<0.01) in the saline, Null-MSCs and Ang-MSCs groups.Compared with the saline group, the escape latency of Water Maze was significantly shortened (all P<0.01) and the times of crossing the original table in 2 minutes were significantly increased (all P<0.01) in the Null-MSCs and Ang-MSCs groups, and the Ang-MSCs group showed the most significant improvement (P<0.05 or P<0.01).Conclusion Ang-1 gene modified MSCs can improve the spatial learning and memory abilities of the cerebral infarction rats.
出处
《山东医药》
CAS
北大核心
2015年第40期4-7,共4页
Shandong Medical Journal
基金
山东省科技发展计划项目(2011GSF11831)
山东省自然基金资助项目(Z2008C06)
关键词
脑梗死
血管生成素-1
骨髓间充质干细胞
腺病毒
基因转染
大鼠
cerebral infarction
angiopoietin-1
bone marrow mesenchymal stem cells
adenovirus
gene transfection
rats
