体外转基因成肌细胞移植对大鼠损伤脊髓细胞凋亡的影响

目的：探讨大鼠脊髓损伤后胚胎脊髓和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对大鼠脊髓细胞凋亡的影响。方法：将动物分为：大鼠脊髓半切洞损伤明胶海绵填充组（A组），大鼠脊髓半切洞损伤应用胚胎脊髓移植组（B组），脊髓半切洞损伤损伤AxCA-BDNF基因转染的成肌细胞移植组（C组）大鼠脊髓半切洞损伤后应用胚胎脊髓和AxCA-BDNF基因转染的成肌细胞移植组（D组）。手术后1、3、7、14、28d应用行为学和电生理检查观察大鼠功能恢复情况，对脊髓损伤区进行细胞凋亡的检测（TUNEL）以及Bcl-2蛋白表达的测定（免疫组化法）。采用计算机图像分析技术，进行定量分析。结果：A、B、C、D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，各组凋亡细胞核为A＞B＞C＞D；Bcl-2免疫反应阳性细胞表达顺序为D＞C＞B＞A，Bcl-2免疫反应阳性细胞的表达与大鼠后肢功能恢复有同样的变化趋势。结论：大鼠胚胎脊髓和体外转基因成肌细胞移植能抑制脊髓损伤后的细胞凋亡。

Effect of Intraspinal Grafting of Adenovirus-Mediated BDNF vitro Transgene Myoblasts Cells and Fetal Spinal Cord on Apoptosis after Spinal Cord Injury

Objective: To investigate the effect of intraspinal grafting of fetal spinal cord (FSC)and vitro transgene myoblasts cells on apoptosis after spinal cord injury(SCI).Methods:Experimental rats were divided into four groups:Spinal cord hemisection injury and received gelfoam pledgets group (group C),Spinal cord hemisection injury combination with transplants of FSC and vitro transgene myoblasts cells to injured site group (group A).Spinal cord hemisection injury with transplants of FSC to injured site group(group B),Spinal cord hemisection injury with transplants of vitro transgene myoblasts cells to injured site group (group D),After operation Tarlov score,somatosensory evoked potentials(SEP),motor evoked potentials(MEP)were examined.Rats were killed at 1,3,7,14 and 28 day.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 measured by immunohistochemistry.The positive cells were quantitative 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,they are A>B>C>D(P<0.05),D>C>B>A(P<0.05)respectively.These increases in Bcl 2 positive cells were paralleled by a significant improvement in neurological function recovery.Conclusion:It indicates that the intraspinal grafting of fetal spinal cord(FSC) and vitro transgene myoblasts cells can inhibit the apoptosis of injured spinal cord.