人脐血间充质干细胞移植对创伤性脑损伤大鼠VEGF分泌及血管新生的影响  

The influence of HUCBMSCs transplantation of on cerebral angiogenesis and secretion of VEGF in rats with traumatic brain injury

作　　者：赵俊暕[1] 陈乃耀[1] 石峻[1] 王沂[1] 孟爱国[1] 韩晓燕[1] 赵辉[2] 

机构地区：[1]河北联合大学附属医院检验科,063000 [2]天津商业大学天津市食品与生物技术重点实验室,30013

出　　处：《中国神经免疫学和神经病学杂志》2013年第4期267-273,共7页Chinese Journal of Neuroimmunology and Neurology

基　　金：河北省2010年医学科学研究重点课题(河北省卫生厅20100131)

摘　　要：目的探讨人脐血间充质干细胞(MSCs)移植对大鼠创伤性脑损伤(TBI)模型的脑保护作用机制。方法大鼠随机分为假手术组、损伤对照组(对照组)和实验组,每组30只。对实验组和对照组采用改良Feeney自由落体方法制作大鼠TBI模型。假手术组只开骨窗,不撞击硬脑膜。造模操作后24h,实验组所有大鼠均经鼠尾静脉注入用Brdu标记的3×106个/mL MSCs(以1mL PBS液溶解)1mL;对照组和假手术组所有大鼠经鼠尾静脉注入等体积的PBS液。3组大鼠均未使用免疫抑制剂。以神经损伤严重缺损评分(NSS)方法评价各组大鼠神经损伤程度;各组大鼠注射后第3、7、14、21、28天,采用HE染色观察脑损伤周边区组织形态学变化,免疫组化方法观察损伤周边区Brdu阳性的MSCs分布、微血管密度及VEGF、VEGF-R2表达,原位杂交方法观察VEGF mRNA表达,并进行比较。结果实验组大鼠均未发现免疫排斥反应。注射后第7、14、21、28天实验组大鼠NSS评分均低于对照组(P<0.05)。光镜下假手术组大鼠脑组织细胞结构基本正常;对照组大鼠脑组织结构破坏,损伤区可见大量坏死细胞,胞核皱缩,细胞结构消失。实验组脑组织坏死范围缩小,细胞结构较对照组完整,核固缩较对照组明显减轻。Brdu阳性的MSCs在实验组大鼠脑损伤周边区明显聚集,4周后仍可见存活MSCs。实验组大鼠脑损伤周边区有较多单个血管内皮细胞和条索状的血管样结构,各时间点实验组MVD值均高于对照组(P<0.01);注射后各时间点实验组在损伤周边区表达细胞因子VEGF、VEGF-R2和VEGFmRNA的细胞计数均较对照组明显增多(P<0.05)。对照组和实验组因子VEGF、VEGF-R2和VEGFmRNA表达水平均分别在注射后第7、3、3天达高峰,此后对照组表达逐渐减弱,实验组第21天、甚至28天因子表达仍维持较高水平。结论在未应用免疫抑制剂的情况下,经尾静脉注射的MSCs可迁移至免疫功能健全的大鼠Objective To study the effects of intravenous administration of human umbilical cord blood mesenchymal stem cells（HUCBMSCs） in traumatic brain injury（TBI） rats,and explore its mechanisms promoting the recovery of neurological function. Methods Healthy male Sprague-Dawley rats were randomly divided into the sham-operated group,the injury control group,and the treated group.There were 30 rats in each group.A modified Feeney free-falling method was used in both the injury control group and the transplantation treated group.In the sham-operated group the rats were only opened bone Windows.The dura mater was not hit.Twenty four hours after modeling,the rats in the treated group were injected with 3×106/mL HUCBMSCs labelled by Brdu suspended with 1 mL PBS via tail vein.The rats in the sham injury group and the control group were injected with an equal volume of PBS.No immunosuppressive agents were used in all groups.The rats were killed at corresponding time points after evaluating the Neurological Severity Score Points（NSS）.HE staining,immunohistochemistry,in situ hybridization were adopted in the rats at different time points after injection（3 d,7 d,14 d,21 d,28 d）,the morphological changes of brain tissue were observed under light microscope and the expression of VEGFmRNA,VEGF and VEGF-R2,as well as angiogenesis situation in temporal parietal cortex with the same phase point in each group after transplantation were detected and compared hetween different groups. Results No transplant-related deaths were observed in the treated group.NSS score showed that since the 7th day of HUCBMSCs injection,neurological function of the treatment group was significantly improved compared with the control group（P0.05）.Under light microscopy,the cellular structure of rat brain was normal in the sham-operated group.Breakdown and shedding of tissue,neuron denaturation,shrunken cell body,piles of cell with condensed nucleus could be seen in the cortex of the control group.Pathological changes of the rats in the tre

关 键 词：人脐血间充质干细胞 创伤性脑损伤 VEGF VEGF-R2 血管新生 

分 类 号：R741.05[医药卫生—神经病学与精神病学]