脑源性神经生长因子转基因细胞移植和大剂量甲基强的松龙对脊髓损伤后细胞凋亡的影响  

Effect of adenovirus-mediated brain derived neurotrophic factor ex vivo transgene myoblasts cells and methylprednisolone on apoptosis after spinal cord injury

作　　者：张强[1] 邹德威[1] 海涌[1] 白克文[1] 

机构地区：[1]解放军306医院骨科全军脊柱外科中心,北京市100101

出　　处：《中国脊柱脊髓杂志》2005年第7期425-428,i0002,共5页Chinese Journal of Spine and Spinal Cord

摘　　要：目的:探讨大鼠脊髓损伤后腺病毒介导的脑源性神经生长因子(AxCA-BDNF)体外转基因成肌细胞移植和静脉内注射大剂量甲基强的松龙(MP)对大鼠脊髓损伤后细胞凋亡的影响。方法:120只Wistar大鼠分为:脊髓挫伤组(A组),脊髓挫伤后AxCA-BDNF基因转染的成肌细胞移植组(B组),脊髓挫伤后静脉内注射大剂量MP治疗组(C组),脊髓挫伤后同时应用AxCA-BDNF和MP组(D组)。手术后1、3、7、14、28d用行为学和电生理检查观察大鼠功能恢复情况,并用计算机图像分析技术对脊髓损伤区细胞凋亡(TUNEL法)和Bcl-2蛋白表达(免疫组化法)进行定量分析。结果:四组中均发现凋亡细胞及Bcl-2蛋白阳性表达细胞,图像分析发现四组凋亡细胞核数为A组>B组>C组>D组;Bcl-2免疫反应阳性细胞表达顺序为D组>C组>B组>A组,大鼠后肢功能恢复和电生理检查也有类似的变化趋势。结论:体外转基因成肌细胞移植和大剂量MP都能抑制大鼠脊髓损伤后的细胞凋亡,促进大鼠后肢功能恢复,两者联合应用具有协同作用。Objective：To investigate the effect of brain derived neurotrophic factor（BDNF） ex vivo transgene myoblasts cells and methylprednisolone（MP） on apoptosis after spinal cord injury（SCI）.Metbod：One hundred and twenty rats were divided into four groups,spinal cord contusion injury（group A）,spinal cord contusion injury with transplantation of ex vivo transgene myoblasts cells into injured site （group B）,spinal cord contusion injury with large dose of MP intravenous injection （group C）,spinal cord contusion injury with transplantation of transgene myoblasts cells to injured site combined with large dose of MP intravenous injection （group D）, after operation.Tarlv score,somatosensory evoked potentials（SEP）,motor evoked potentials（MEP） were examined at 1,3,7,14 and 28 day post operation.The apoptosis of spinal slice from the injured spinal cord were examined by the methods of the terminal deoxynucleotidal transferase-mediated DUTP-biotin nick end labeling （TUNEL） reaction and the expression of Bcl-2 was tested by immunohistochemistry.The positive cells were quantitatively analyzed by a computer image analysis system.Result：The number of apoptosis cells and the strong expression of Bcl-2 protein expression were found in four groups,the group ranking was A〉B〉C〉D （P〈0.05） as for the number of apoptosis cells and D〉C〉B〉A （P〈0.05） as for the expression of Bcl-2.These increases in Bcl-2 positive cells were paralleled by a significant improvement in neurological function and the peak latencies of early waves in SEPs and MEPs recovery.Conclusion：Intraspinal grafting of transgene myoblasts cells and MP intravenous injection respectively can inhibit the cell apoptosis of injured spinal cord as well as promote the recovery of neural function of adult rat injured spinal cord.Combined intraspinal grafting of transgene myoblasts cells with MP intravenous injection have a coordination effect.

关 键 词：脊髓损伤 转基因 脑源性神经生长因子 甲基强的松龙 细胞凋亡 

分 类 号：R322.81[医药卫生—人体解剖和组织胚胎学] R977.1[医药卫生—基础医学]