摘要
背景:动物实验显示人脐带间充质干细胞移植具有显著的神经保护作用,其移植途径包括经静脉、动脉、腰穿、脑立体定向移植和侧脑室移植多种,均存在不足。目的:探讨通过鼻腔移植人脐带间充质干细胞修复新生大鼠缺氧缺血性脑损伤的可行性和有效性。方法:取90只7 d龄SD大鼠(汕头大学医学院实验动物中心提供),随机分3组,每组30只:假手术组分离左颈总动脉后缝合皮肤;模型组采用结扎切断左颈总动脉联合缺氧仓的方法中制备缺氧缺血性脑损伤模型;实验组采用同样方法制备缺氧缺血性脑损伤模型,同时鼻腔滴入绿色荧光蛋白标记的人脐带间充质干细胞。建模后3 d取脑组织,进行尼氏染色、TUNEL染色及免疫荧光染色;建模后14,29 d,水迷宫实验检测大鼠记忆、学习能力,14d水迷宫实验完成后行脑组织TUNEL染色。动物实验经深圳大学医学部实验动物伦理委员会批准(伦理批准号:2016-121)。结果与结论:①尼氏染色:模型组大脑皮质及海马CA1区神经元排列均不规则,尼氏小体数量少;实验组大鼠大脑皮质及海马CA1区神经元排列较模型组规则,尼氏小体数量较多;②TUNEL染色:建模后3,14 d,模型组凋亡细胞数多于假手术组(P <0.05),实验组凋亡细胞数少于模型组(P <0.05);③免疫荧光染色:实验组脑组织中可见绿色标记的人脐带间充质干细胞,主要分布于损伤侧海马及大脑皮质区域,非损伤侧脑组织偶可见绿色标记细胞,未见绿色荧光与GFAP或NSE染色双阳性细胞;④实验组建模后14,29 d的平均逃逸潜伏期均低于模型组(P <0.05);⑤结果表明:经鼻腔移植人脐带间充质干细胞修复新生大鼠缺氧缺血性脑损伤,可减少神经细胞凋亡,改善大鼠记忆学习能力。
BACKGROUND:Human umbilical cord mesenchymal stem cells exert significant neuroprotective effects in animals,including reducing neuronal apoptosis and promoting neuronal regeneration.Current approaches to transfer these cells include transvenous,arterial,lumbar puncture,brain stereotactic transplantation and lateral ventricle transplantation,but all of them are insufficient.OBJECTIVE:To investigate the feasibility and effectiveness of nasal transplantation of human umbilical cord mesenchymal stem cells in the treatment of hypoxic-ischemic brain damage in neonatal rats.METHODS:Ninety 7-day-old Sprague-Dawley rats(provided by the Experimental Animal Center of Shantou University Medical College)were randomly divided into three groups(n=30 per group):in sham operation group the left common carotid artery was only separated and then sutured;in model group,an animal model of hypoxic-ischemic brain damage was prepared through ligation and occlusion of the left common carotid artery combined with hypoxic chamber;in experimental group,the animal model was prepared as described in the model group,and then green fluorescent protein-labeled human umbilical cord mesenchymal stem cells were instilled into the nasal cavity of model rats.Brain tissue was taken 3 days after modeling,and Nissl staining,TUNEL detection,and immunofluorescence staining were performed.The memory and learning abilities of the rats were detected by water maze at 14 and 29 days after modeling.After 14-day water maze test,TUNEL staining of the brain tissue was performed.The animal experiment was approved by the Experimental Animal Ethics Committee of Shenzhen University Health Science Center with the approval No.2016-121.RESULTS AND CONCLUSION:(1)Nissl staining:Neurons in the cerebral cortex and hippocampal CA1 area of the model group were irregularly arranged,and there were few Nissl bodies.Compared with the model group,the neurons in the cerebral cortex and hippocampal CA1 area were arranged more regularly in the experimental group,and there were more Ni
作者
杨汉华
田树凤
谢丽春
陈运彬
马廉
Yang Hanhua;Tian Shufeng;Xie Lichun;Chen Yunbin;Ma Lian(Department of Neonatology,Medical and Child Health Care Hospital of Pingshan District,Affiliated Maternal and Child Health Hospital of Shenzhen University,Shenzhen 518122,Guangdong Province,China;Department of Infectious Diseases,Shenzhen Children's Hospital,Shenzhen 518038,Guangdong Province,China;Department of Hematology Oncology,Shenzhen Children's Hospital,Shenzhen 518038,Guangdong Province,China;Guangdong Maternal and Child Health Hospital,Guangzhou 510010,Guangdong Province,China;Institute of Pediatrics,Shenzhen Children's Hospital,Shenzhen 518038,Guangdong Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2019年第21期3386-3391,共6页
Chinese Journal of Tissue Engineering Research
基金
深圳市知识创新计划项目(JCYJ20160429141742207)
项目负责人:杨汉华
国家自然科学基金(81671525)
项目负责人:马廉~~
关键词
干细胞
脐带干细胞
间充质干细胞
脐带间充质干细胞
移植
缺氧缺血性脑损伤
鼻腔给药
水迷宫实验
stem cells
umbilical cord blood stem cells
mesenchymal stem cells
umbilical cord blood mesenchymal stem cells
transplantation
hypoxic-ischemic brain damage
intranasal delivery
water maze test
