人脐带间充质干细胞对新生大鼠脑白质损伤后神经胶质细胞及神经行为学的作用  

Effects of human umbilical cord mesenchymal stem cells intervention on glial cells and neurobehavior in the newborn rat model of white matter damage

作　　者：摆翔[1] 王诗雨[1] 张宁[1] 朱丽华[1] 蒋犁[1] 李伟 

机构地区：[1]东南大学医学院儿科,南京210009 [2]江苏省干细胞工程技术研究中心

出　　处：《中华围产医学杂志》2014年第9期614-620,共7页Chinese Journal of Perinatal Medicine

基　　金：国家自然科学基金（81370739）

摘　　要：目的 探讨人脐带间充质干细胞（human umbilical cord mesenchymal stem cells,HUcMSCs）移植对脑白质损伤（white matter damage,WMD）新生大鼠脑内少突胶质细胞及活化小胶质细胞的影响,以及新生大鼠远期神经行为学的改变. 方法 72只3日龄Sprague-Dawley大鼠行左侧颈总动脉结扎,并吸入6％O2和94％N2的混合气体4h,制成WMD模型.死亡12只,剩余60只随机分为实验组和对照组各30只.大鼠出生后0～24 h计为第1天,实验组大鼠于第3、4和5天每天腹腔注射HUcMSCs 1×106个（0.05 ml）,对照组大鼠于同时间点腹腔注射磷酸盐缓冲液0.05 ml.2组均于第10和21天分别取8只大鼠,免疫组织化学方法检测左侧脑组织中髓鞘碱性蛋白（myelin basic protein,MBP）和单核巨噬细胞抗原（ectodermal dysplasia,ED）-1染色阳性细胞.2组均于第10和21天分别取3只大鼠,Western印迹技术检测MBP和ED-1蛋白表达.2组分别取8只大鼠测量体重,并于第28天行神经行为学评估.两样本均数比较采用t检验. 结果 实验组第10和21天MBP染色阳性细胞数[（11.8±4.1）和（23.8±8.1）个]均高于对照组[（6.7±3.1）和（11.5±5.8）个,t值分别为2.81和3.49,P值均＜0.05],ED-1染色阳性细胞数[（20.8±3.4）和（19.1±2.8）个]均低于对照组[（32.8±4.2）和（29.5±5.2）个,t值分别为6.23和4.94,P值均＜0.01].实验组第10和21天MBP的表达量（1.3±0.1和1.1±0.1）均高于对照组（0.8±0.0和0.6±0.1,t值分别为7.53和6.68,P值均＜0.01）,ED-1的表达量（0.6±0.1和0.4±0.1）均低于对照组（1.0±0.1和0.8±0.1,t值分别为3.90和4.90,P值均＜0.01）.实验组第7、10、14、21和28天体重分别为（15.0±1.2）、（16.6±0.9）、（27.0±1.6）、（44.2±2.3）和（68.1±4.2）g,均高于对照组[分别为（12.7±1.6）、（13.5±2.0）、（23.6±1.9）、（38.4±0.9）和（60.0±4.2）g,t值分别为-3.11、-3.97、-3.67、-6.52和-3.72,P值均＜0.05）.第28天神经行为学测试：实验Objective To investigate the alterations in oligodendrocyte and microglia and changes in neurobehavior after human umbilical cord mesenchymal stem cells （HUcMSCs） intervention on the newborn rat model of white matter damage （WMD） induced by hypoxia-ischemia.Methods After the operation of left common carotid artery ligation and 4 h hypoxia （6% O2 and 94% N2）,twelve three-day-old Sprague-Dawley rats died and the remaining sixty rats were randomly divided into the experimental group and the control group.The first day was 0 to 24 h after birth.Rats of the experimental group were intraperitoneally injected the fourth generation HUcMSCs 1 × 106 （0.05 ml） on the third,fourth and fifth day respectively.At the same time,rats of the control group were intraperitoneally injected phosphate buffer （0.05 ml）.Eight rats of each group were executed on the tenth and twenty first day respectively to detect the number of cells positive for myelin basic protein （MBP） and ectodermal dysplasia-1 （ED-1） staining by immunohistochemistry.Three rats of each group were executed on the tenth and twenty first day respectively to detect MBP and ED-1 expression by western blot.Eight rats of each group were weighed and underwent the neurobehavioral evaluation on the twenty eighth day.Data were analyzed using t test.Results On the tenth and twenty first day,the numbers of MBP-positive cells in the experimental group （11.8 ± 4.1 and 23.8± 8.1） were significantly higher than those in the control group （6.7±3.1 and 11.5 ± 5.8,t=-2.81 and 3.49,both P〈0.05） ; and the numbers of ED-1 positive cells in the experimental group （20.8 ± 3.4 and 19.1 ± 2.8） were significantly lower than those in the control group （32.8±4.2 and 29.5±5.2,t=6.23 and 4.93,both P〈0.01）.On the tenth and twenty first day,MBP expressions in the experimental group （1.3 ± 0.1 and 1.1 ± 0.1） were higher than those in the control group （0.8±0.0 and 0.6±0.1,t=-7.53 and 6.68,both P〈0.01） ; and the ED-1 expressions in

关 键 词：脑白质病 脐带 间充质干细胞 少突神经胶质 小神经胶质细胞 间质干细胞移植 

分 类 号：R722.1[医药卫生—儿科]