脐血源神经干细胞移植治疗大鼠脊髓损伤及其差异基因表达  

Umbilical cord blood neural stem cells transplantation for treatment of spinal cord injury in rats and differential expression gene

作　　者：吴月奎[1] 王尚武[1] 孙起军[1] 霍铁军[1] 马建华[2] 郭海若[2] 秦家振[2] 戴宜武[2] 徐如祥[2] 

机构地区：[1]首都医科大学密云教学医院神经外科,北京101500 [2]北京军区总医院附属八一脑科医院,北京100700

出　　处：《中华神经医学杂志》2014年第8期772-777,共6页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金（81271391,30973225）

摘　　要：目的 评估脐血源神经干细胞（UCBNSCs）移植治疗脊髓损伤（SCI）后大鼠后肢功能恢复情况,并通过基因芯片筛选出移植治疗后对照组和实验组差异表达基因,分析相关基因与损伤脊髓功能恢复的关系,以期为SCI的基因治疗打下基础. 方法 50只健康雌性SD大鼠按随机数字表法分为假手术组（n=10）、磷酸盐缓冲液（PBS）注射组（n=20）、UCBNSCs移植组（n=20）.假手术组仅在显微镜下行T9-11椎板打开,不做其他处理.后2组打开T9-11椎板后横断脊髓,制作大鼠脊髓全断模型.1周后UCBNSCs移植组行UCBNSCs（1×10^9/L细胞悬浮液10μL）脑室穿刺移植,PBS注射组用PBS代替.移植术后1d、1周、2周、4周、6周、8周分别对3组大鼠行血脑屏障（BBB）行为评分.第8周分别取PBS注射组和UCBNSCs移植组各7只SD大鼠,取得SCI处组织样本,行基因芯片分析,筛选出差异表达基因. 结果 PBS注射组和UCBNSCs移植组造模后均出现了瘫痪、尾部活动障碍及排尿功能障碍等.随着时间的延长,2组后肢功能均有所恢复,在术后2周、4周、6周和8周2组间差异具有统计学意义（P＜0.05）.芯片分析筛选出24个差异表达基因,其中表达上调有20个,表达下调有4个. 结论 UCBNSCs移植治疗大鼠SCI有确切疗效,通过基因芯片分析推测其可能通过分化为神经元、增加损伤微环境中神经营养因子表达、抑制炎症反应和促进损伤局部的氧供等方式发挥作用.Objective To assess the recovery of hind limbs function in rats with spinal cord injury （SCI） after being treated with umbilical cord blood neural stem cells （UCBNSCs） transplantation,and analyze the relationship between the differentially expressed genes identified by gene chips and functional recovery of the spinal cord to lay foundation for gene therapy of spinal cord injury.Methods Fifty healthy female SD rats were randomly divided into sham-operated group （n=10）,PBS injection control group （n=20） and UCBNSCs transplantation group （n=20）.Only opening the T9-11 vertebral plate was performed in sham-operated group under microscope,while the spinal cords of PBS injection control group and UCBNSCs transplantation group were cut offafter opening the T9-11 vertebral plate to induce rat models of spinal cord transection,UCBNSCs transplantation was performed through ventricular puncture （1 × 109/L cell suspension 10 μL） one week after the model making.Blood brain barrier （BBB） behavior scale of three groups was carried out 1 d and 1,2,4,6 and 8 w after transplantation.Seven SD rats from PBS control group and UCBNSCs transplantation group were sacrificed to get spinal cord injury tissue samples at 8 w after transplantation; gene chip analysis was used to identify the differentially expressed genes.Results The hind limbs function of SD rats did not appear obvious damage in sham-operated group.Symptoms of paralysis,tail activity obstacle and urination dysfunction appeared in PBS injection control group and UCBNSCs transplantation group.As time went on,the function of hind limbs slowly recovered：1 d and 1 w after transplantation,BBB scale scores showed no statistical difference between PBS injection control group and UCBNSCs transplantation group （P〉0.05）; statistical significance appeared at 2,4,6 and 8 w after transplantation between the two groups （P〈0.05）.Twenty-four differentially expressed genes were identified,including 20 being up-regulated and 4 being down-regulated.Co

关 键 词：脐血源神经干细胞 脊髓损伤 差异基因 

分 类 号：R651.2[医药卫生—外科学]