脂肪干细胞旁分泌提取物对新生大鼠脑白质损伤的影响  

Neuroprotective effect of paracrine extracts derived from human adipose stem cells on white matter injury of neonatal rats

作　　者：黄超[1,3] 杨印祥[1] 汪兆艳[1] 王倩[1] 陈敬国[2] 栾佐[1] 

机构地区：[1]海军总医院儿科,北京100048 [2]中山大学附属中山市人民医院新生儿科 [3]中山大学

出　　处：《中国新生儿科杂志》2017年第2期144-149,共6页Chinese Journal of Neonatology

基　　金：科技部国际合作专项项目（2012DFA30880）

摘　　要：目的 探讨脂肪干细胞旁分泌提取物对新生大鼠脑白质损伤的影响,比较小脑延髓池注射途径与颈总静脉注射途径的疗效.方法 选择3日龄新生SD大鼠73只制作早产儿脑白质损伤模型,建模后24 h将大鼠随机分为实验组46只和对照组27只.实验组再随机分为脑室组和静脉组各23只,分别从小脑延髓池和颈总静脉注射脂肪干细胞旁分泌提取物;对照组再随机分为脑室对照组15只和静脉对照组12只,分别从小脑延髓池和颈总静脉注射等量生理盐水.注射ld后脑室组和静脉组各随机选择3只大鼠行脑组织冰冻切片生物素荧光染色观察提取物分布;3d后脑室组、静脉组和脑室对照组各随机选择3只大鼠行苏木精-伊红（HE）染色观察脑组织病理形态改变,7d后4个亚组各随机选择7只大鼠检测髓鞘碱性蛋白（MBP）表达;28 d后剩余大鼠行神经行为学实验（2个对照组合并分析）.实验过程中如有大鼠死亡,补足各组大鼠只数.结果 建模后24 h内无大鼠死亡,分组后的实验过程中实验组死亡3只,对照组死亡4只,均予以补充.实验组两种注射途径下均可见脂肪干细胞旁分泌提取物迁移至脑部病灶区域,且小脑延髓池注射途径脂肪干细胞旁分泌提取物入脑数量多于颈总静脉注射途径.HE染色结果显示,脑室对照组脑白质区域细胞损伤严重,发生大范围梗死,可见微小梗死灶;实验组损伤较轻,其中脑室组较静脉组胼胝体结构更完整.脑室组和静脉组MBP半定量评分分别低于脑室对照组和静脉对照组[（0.7±0.3）分比（3.4±0.4）分,（1.7±0.3）分比（3.3±0.3）分],脑室组低于静脉组,差异有统计学意义（P＜0.05）.脑室组和静脉组悬吊实验和旷场实验评分均高于对照组[（2.9±0.2）分、（1.9±0.2）分比（1.3±0.1）分,（11.6±1.1）分、（10.4±1.1）分比（9.0±1.3）分],脑室组高于静脉组,差异有统计学意义（P＜0.05）.结论 脂肪Objective To explore the effect of paracrine extracts derived from human adipose stem cells on white matter injury of neonatal rats and to compare the difference of therapeutive effect between the cerebellum medulla oblongata pool injection and the jugular vein injection.Method A total of 73 three-day-old SD rats were chosen to establish the model of white mater injury.After 24 hours,the 73 rats were randomized into the experimental group （n =46） and the control group （n =27）.Then the experimental group was reclassified into ventricular group （n =23） and intravenous group （n =23）.In the ventricular group,the paracrine extracts of human adipose stem cells was injected locally into the cerebellum medulla oblongata pool injection,while the extracts was injected into the jugular vein in the intravenous group.The control group was reclassified ventricular control group （n =15） and intravenous control group （n =12）,and equivoluminal saline was injected the same way as the experimental group.Frozen sections of the brain tissue from 3 rats of each experimental group one day after injection were stained with fluorescein-conjugated streptavidin to study the distribution of the extracts.The brain tissue of 3 rats from each subgroup 3 days after injection were stained with hematoxylin eosin （HE） to observe the pathomorphological changes.While 7 days later,myelin basic protein （MBP） of white matter which was obtained from 7 rats of each group was detected by immunofluorescence staining.28 days after injection,the remaining rats were assessed by neurobehavior tests.For the rats that died during the experiment,the same number of the rats would be substituted in this study.Result The paracrine extracts were found to transfer to the brain lesion area,and the amount of the extracts was more in the ventricular group.The results of the HE staining showed that the white matter injury was more severe in the ventricular control group,and extensive area of infarction were found in this group.White matter

关 键 词：脂肪因子类 干细胞 脑白质损伤 大鼠 

分 类 号：R742[医药卫生—神经病学与精神病学]