两种干细胞经鼻移植治疗大鼠早产儿脑白质损伤  

Two kinds of stem cell nasal transplantation for treating white matter injury in premature rat infants

作　　者：臧静 栾佐 王倩 杨印祥 汪兆艳 吴尤佳[3] 郭爱松[4] Zang Jing;Luan Zuo;Wang Qian;Yang Yinxiang;Wang Zhaoyan;Wu Youjia;Guo Aisong(Medical School of Nantong University,Nantong 226001,Jiangsu Province,China;Department of Pediatrics,The Sixth Medical Center of Chinese PLA General Hospital,Beijing 100048,China;Department of Pediatrics,Affiliated Hospital of Nantong University,Nantong 226001,Jiangsu Province,China;Department of Traditional Chinese Medicine,Affiliated Hospital of Nantong University,Nantong 226001,Jiangsu Province,China)

机构地区：[1]南通大学医学院,江苏省南通市226001 [2]中国人民解放军总医院第六医学中心儿科,北京市100048 [3]南通大学附属医院儿科,江苏省南通市226001 [4]南通大学附属医院中医科,江苏省南通市226001

出　　处：《中国组织工程研究》2021年第1期101-107,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家重点研发计划(2017YFA0104200),项目负责人:栾佐。

摘　　要：背景:干细胞移植对神经系统疾病具有显著的神经保护作用,目前的移植方式如动静脉移植、脑立体定位移植等均不适用于早产儿临床应用。目的:探讨人脐带间充质干细胞和人神经干细胞经鼻移植治疗大鼠早产儿脑白质损伤的可行性。方法:采用人脐带组织制备人脐带间充质干细胞,人胚胎脑组织制备人神经干细胞,通过Transwell法测定两种细胞体外迁移情况;将40只3 d龄SD大鼠随机分为假手术组、模型对照组、人脐带间充质干细胞移植组和人神经干细胞移植组,每组10只;除假手术组,其余各组行右颈总动脉离断并缺氧90 min制备大鼠早产儿脑白质损伤模型。移植组于造模后3 d经鼻滴注总量为1×10^6细胞,每侧鼻孔滴注5×10^5,双侧鼻孔分别滴注1次。造模后第7天采用免疫荧光染色检测脑白质MBP表达来鉴定脑白质损伤情况;移植后24 h抗HuNu免疫组化方法检测人脐带间充质干细胞迁移情况,CM-Dil荧光标记法检测人神经干细胞迁移情况。结果与结论:①造模后第7天早产儿脑白质损伤模型鼠患侧扣带、胼胝体和外囊与正常侧对比,MBP阳性面积减少(P<0.05),表明造模成功;②体外实验发现人神经干细胞和人脐带间充质干细胞迁移率一样;③经鼻移植细胞后24 h,人脐带间充质干细胞迁移至正常侧和损伤侧皮质、胼胝体和海马区域,人神经干细胞迁移至损伤侧皮质、胼胝体和海马区域,且人脐带间充质干细胞迁移量相对人神经干细胞较多;④结果表明,经鼻移植24 h后,人脐带间充质干细胞能够存活并向正常侧和损伤侧迁移,人神经干细胞能够存活并向损伤侧迁移,且人脐带间充质干细胞迁移较人神经干细胞体内迁移更广泛。BACKGROUND:Stem cell transplantation has a significant neuroprotective effect on neurological diseases.Current transplantation methods such as arteriovenous transplantation and brain stereotactic transplantation are not suitable for clinical application in preterm infants.OBJECTIVE:To explore the feasibility of nasal transplantation of human umbilical cord mesenchymal stem cells and human neural stem cells for the treatment of white matter injury in premature rat infants.METHODS:Human umbilical cord mesenchymal stem cells were prepared from human umbilical cord tissue,and human neural stem cells were prepared from human embryonic brain tissue.In vitro migration of two kinds of cells was assessed by Transwell method.Forty 3-day-old Sprague-Dawley rats were randomly divided into sham operation group,model control group,human umbilical cord mesenchymal stem cell transplantation group and human neural stem cell transplantation group,with 10 rats in each group.Rats in all groups except the sham operation group were treated with right common carotid artery ligation and hypoxia for 90 minutes to establish a rat model of white matter injury in the preterm infant.Totally 1×10^6 cells were delivered intranasally in the transplantation group at 3 days after injury.Each nostril was infused with 5×10^5,and each nostril was infused once.On day 7 after injury,MBP immunofluorescence staining was used to detect the expression of myelin basic protein in the white matter of the brain to identify the damage of the white matter injury model.At 24 hours after transplantation,human umbilical cord mesenchymal stem cell migration was detected by anti-HuNu immunohistochemical method and human neural stem cell migration was detected by CM-Dil fluorescent labeling method.RESULTS AND CONCLUSION:(1)On day 7 after modeling,compared with the normal side,the positive area of MBP decreased in cingulate band,corpus callosum and external capsule of the affected side in the model of brain white matter injury in preterm infants(P<0.05),indicating a

关 键 词：干细胞 移植 人脐带间充质干细胞 人神经干细胞 鼻 早产儿 脑白质损伤 大鼠 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]