中国儿童保健杂志 ›› 2017, Vol. 25 ›› Issue (1): 32-36.DOI: 10.11852/zgetbjzz2017-25-01-10

• 基础科研论著 • 上一篇    下一篇

人脐血间充质干细胞移植联合mNGF治疗脑瘫大鼠的实验研究

王丽敏1,谭振香2,霍明霞1,郭佳丽3,袁晓辉1,袁博洋1,王文娟1,张雪玲1   

  1. 1 佳木斯大学附属第一医院儿科、中医科,黑龙江 佳木斯 154003;
    2 绥芬河市妇幼保健院,
    黑龙江 牡丹江 157399;
    3 徐州医学院附属连云港市第一人民医院儿科,江苏 连云港 222000
  • 收稿日期:2016-05-18 出版日期:2017-01-10 发布日期:2017-01-10
  • 通讯作者: 谭振香,E-mail:tzx_0453899@126.com
  • 作者简介:王丽敏(1964-),女,黑龙江人,主任医师,教授,主要从事小儿心肾及康复研究。
  • 基金资助:
    佳木斯大学科学技术项目(13Z1201541)

Research of transplantation of mesenchymal stem cells from umbilical cord blood combined with mNGF in the treatment of cerebral palsy rats.

WANG Li-min1,TAN Zhen-xiang2,HUO Ming-xia1,GUO Jia-li3,
YUAN Xiao-hui1,YUAN Bo-yang1,WANG Wen-juan1,ZHANG Xue-ling1.   

  1. 1 Department of Pediatrics and Traditional Chinese Medicine,the First Hospital Affiliated to Jiamusi University,Jiamusi,Heilongjiang 154003,China;
    2 Department of Pediatrics,Maternal and Child Care Service Centre of Suifenhe,Mudanjiang,Heilongjiang 157399,China;
    3 Department of Pediatrics,Lianyungang First People's Hospital Affiliated to Xuzhou Medical College,Jiangsu,Lianyungang 222000,ChinaCorresponding author:TAN Zhen-xiang,E-mail:tzx_0453899@126.com
  • Received:2016-05-18 Online:2017-01-10 Published:2017-01-10
  • Contact: TAN Zhen-xiang,E-mail:tzx_0453899@126.com

摘要: 目的 研究经侧脑室内注入人脐血间充质干细胞(hUCB-MSCs)联合鼠神经生长因子(mNGF)对脑瘫大鼠治疗效果。方法 采集足月新生儿脐带血100 ml,分离、培养、纯化人脐血间充质干细胞并进行检测和鉴定。将七日龄SD大鼠100只随机分为正常组10只、实验组(缺血缺氧性脑瘫模型)90只,实验组随机分为5组脑瘫模型组(CP组)、磷酸盐缓冲溶液(PBS)移植组、mNGF移植组、hUCB-MSCs移植组及hUCB-MSCs联合mNGF移植组。造模成功后1周,将hUCB-MSCs、mNGF分别注入左侧侧脑室,假移植组注射PBS液,移植后第1周、2周、3周,观察5溴-2脱氧尿核苷(BrdU)标记的脐血MSCs的存活、迁移,移植后第4周,对每组大鼠的神经行为学能力进行评估并检测大鼠脑组织胶质纤维酸性蛋白抗体(GFAP)和神经元特异性烯醇化酶(NSE)的表达及观察脑组织的形态学变化。结果 1)人脐血MSCs主要分布在左侧侧脑室、额叶、顶叶皮质及白质,随时间进展,向四周脑组织转移,Brdu、NSE、GFAP阳性细胞逐渐减少,差异有统计学意义(P<0.05);2)行为学评价与模型组及假移植组比较,单移植组、联合移植组的足印潜伏期、迷宫逃避潜伏期缩短,足印重复间距及后足滑脱次数减小,联合移植组优于单移植组,差异有统计学意义(P<0.05);3)HE染色MSCs和mNGF单移植组大鼠细胞形态及脑组织结构破坏较脑瘫组有所改善;联合移植组恢复情况更好,细胞形态恢复较单移植组好。结论 外源性脐血MSCs联合mNGF经侧脑室移植,可改善脑瘫大鼠的远期行为及脑损害。

关键词: 脑瘫, 疗效, 人脐血间充质干细胞, 侧脑室内移植, 鼠神经生长因子

Abstract: Objective To study the treatment effect of human umbilical cord blood mesenchymal stem cells(hUCBMSCs) combined with mouse nerve growth factor (mNGF) to the cerebral palsy rats through lateral intraventricular transplantation. Methods 100 ml cord blood from full-term newborn was collected and to separate,culture and purify hUCBMSCs then detect and identify them.100 seven-day-old SD rats were randomly divided into 10 normal group and 90 experiment group(cerebral palsy produced by ischemia and hypoxia ) while the experiment group were split into cerebral palsy (CP) model group,phosphate buffer solution (PBS) transplantation group,mNGF transplantation group,human umbilical cord blood-derived mesenchymal stem cells transplantation group,and human umbilical cord blood-derived mesenchymal stem cells combined with mouse nerve growth factor transplantation group.After 1 w modeling success MSCs was injected,mNGF and PBS into Lateral ventricles.After transplantation in 1 w,2 w and 3 w,the survival and migration of MSCs with 5 bromine -2 (BrdU) labeled were observed.In the fourth week,the neural behavior ability of rats were studied and the rat brain tissues were taken to detect the expression of its GFAP and NSE. Results 1)The survival,migration of hUCBMSCs mainly distributed at the site of the left lateral ventricle,frontal lobe,parietal cortex and white matter.Along with the time going,its migration was to the surrounding brain tissue.BrdU,NSE,GFAP positive cells decreased gradually,and the difference was statistically significant (P<0.05).Compared with model group and sham transplantation,the footprints latency and maze escape latency of single transplantation groups and combined transplantation group were shorten,footprints repeat spacing and foot slippage number decreasing as well,and transplantation group was better than that of single transplantation groups,the difference was significant (P<0.05);2)HE staining:the cell morphology and brain tissue damage in the single transplantation group were better than that of the cerebral palsy group,and the recovery of the combined transplantation group was better than that of the single tran-splantation group. Conclusion Through intraventricular transplantation,exogenous MSCs transplantation combined with mNGF can improve the long-term behavior and brain damage in cerebral palsy rats.

Key words: cerebral palsy, efficacy, human umbilical cord blood mesenchymal stem cells, lateral intraventricular transplantation, mouse nerve growth factor

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