黄芩苷协同人脐血间充质干细胞静脉移植治疗幼鼠缺氧缺血性脑损伤  

Intravenous transplantation of human umbilical blood mesenchymal stem cells and Baicalin for treating hypoxic-ischemic brain damage in neonatal rats

作　　者：颜小华[1] 徐昕[1] 陈启文[1] 黄涛[1] 管让宪 

机构地区：[1]江西医学院第一附属医院儿科,江西省南昌市330006 [2]江西省儿科医院内科,江西省南昌市330006

出　　处：《中国组织工程研究与临床康复》2009年第49期9650-9654,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:最近研究证实黄芩苷对多种脑损伤具有广泛的神经系统保护作用。目的:探讨中药单体黄芩苷干预并协同人脐血间充质干细胞静脉移植治疗幼鼠缺氧缺血性脑损伤的可行性。设计、时间及地点:随机对照动物实验,于2007-02/2008-01在南昌大学第一附属医院神经内科实验室完成。材料:健康孕妇足月顺产儿的脐带血10份,取自南昌大学一附院产科。清洁级7d龄SD新生鼠85只,随机分为正常对照组15只、模型组20只、细胞移植组25只、细胞移植+黄芩苷组25只。方法:采用明胶沉降+密度梯度离心法分离人脐血单个核细胞,体外扩增至第5代用于移植,使用前6～12h行DAPI标记。模型组、细胞移植组、细胞移植+黄芩苷组新生鼠建立缺氧缺血性脑损伤模型,空白对照组不作任何处理。分别于造模后2,3,4,5周,细胞移植组、细胞移植+黄芩苷组经尾静脉注射DAPI标记好的人脐血间充质干细胞悬液5～10μL/g,细胞浓度为1×109L-1。从细胞移植第1天起,细胞移植+黄芩苷组动物经腹腔注射120mg/kg黄芩苷,1次/d,连续3d。主要观察指标:脑组织病变情况,脑组织DAPI阳性细胞数,脐血间充质干细胞移植后定位情况。结果:移植后4周,细胞移植+黄芩苷组脑组织病变率明显低于模型组、细胞移植组(P<0.05)。与细胞移植组比较,第1,2,4周细胞移植+黄芩苷组DAPI阳性细胞数均显著增加(P<0.01)。造模后第3周开始细胞移植的病灶脑组织,在黄芩苷的协同作用下可见大量DAPI阳性细胞集中分布于病灶区周围,与宿主脑整合,没有明显的界限,而对侧非缺血区很少见到DAPI阳性的脐血间充质干细胞分布;造模后第4,5周开始细胞移植的病灶脑组织,其DAPI阳性细胞明显减少。结论:选择缺氧缺血性脑损伤后第3周进行细胞移植效果较佳,黄芩苷能够协同人脐血间充质干细胞有效地透过血脑屏障,在病灶脑组织周围迁移、扩散、整合BACKGROUND：Present studies have verified that Baicalin has protective effects on various brain damage in the nervous system.OBJECTIVE：To study the possibility of intravenous transplantation of human umbilical blood mesenchymal stem cells （hUBMSCs） and Baicalin after hypoxic-ischemic brain damage （HIBD） in neonatal rats.DESIGN,TIME AND SETTING：The randomized controlled animal study was performed at the Laboratory of the Department of Neurology,First Affiliated Hospital,Nanchang University from February 2007 to January 2008.MATERIALS：Totally 10 umbilical blood samples from healthy full-term pregnant women were obtained from the Department of Obstetrics,First Affiliated Hospital,Nanchang University.A total of 85 clean Sprague Dawley neonatal rats aged 7 days were randomly assigned to a normal control group （n=15）,a model group （n =20）,a cell transplantation group （n =25）,a cell transplantation ＋ Baicalin group （n =25）.METHODS：The umbilical blood mononuclear cells were isolated by the gelatin sedimentation ＋ density gradient centrifugation method,and amplified in vitro.Cells at the fifth passage were used for transplantation.Cells were labeled by DAPI at 6-12 hours before use.Neonatal rats in the model,cell transplantation and cell transplantation ＋ Baicalin groups were used to establish HIBD models.Rats in the blank control group were left intact.At 2,3,4,5 weeks following model induction,rats in the cell transplantation and cell transplantation ＋ Baicalin groups were injected with DAPI-labeled hUBMSCs （5-10 μL/g） via caudal vein at the density of 1×10^9/L.From the first day of transplantation,rats in the cell transplantation ＋ Baicalin group were injected with 120 mg/kg Baicalin via intraperitoneal injection,once a day,for three successive days.MAIN OUTCOME MEASURES：The following parameters were measured：brain tissue lesion,DAPI-positive cell number,location of hUBMSCs following transplantation.RESULTS：Lesion rate of brain tissue was significantly lower in the

关 键 词：黄芩苷 脐血间充质干细胞 移植 缺氧缺血性脑损伤 幼鼠 

分 类 号：R394.2[医药卫生—医学遗传学]