胶质细胞源性神经营养因子和肿瘤坏死因子可溶性受体基因修饰的神经干细胞联合移植治疗脊髓损伤的实验研究

Combined transplantation of neural stem cells transferred with adenoviral GDNF and sTNFR promotes recovery of spinal cord injury

目的探讨胶质细胞源性神经营养因子（GDNF）和肿瘤坏死因子可溶性受体（sTNFR）腺病毒感染的人神经干细胞移植治疗脊髓损伤模型的可行性。方法先包装和扩增含GDNF和sTNFR基因的重组腺病毒，然后用该腺病毒感染人神经干细胞，再将这些神经干细胞移植到脊髓损伤局部。实验动物与分组：雄性sD大鼠，A组：hGDNF和sTNFR转基因神经干细胞移植组；B组：空病毒转基因神经干细胞移植组；c组：神经干细胞移植组；D组：只横断脊髓，不作移植；E组：对照组，只咬除棘突和椎板，不横断脊髓，不做其他处理；F组：不作任何处理。免疫组化检测外源基因的局部表达，最后进行组织学检查和BBB功能评分。结果免疫组化显示局部有外源基因表达，A组BBB功能评分优于C组，但组织学结果未见明显优势；这两组BBB评分显著优于阴性对照组，组织形态也有明显改善。结论腺病毒介导的GDNF和sTNFR基因修饰的神经干细胞移植是一种较好的脊髓损伤治疗方法。

Objective To study the effect of combined transplantation of neural stem cells transferred with adenoviral glial cell line-derived neurotrophic factors （GDNF） and soluble tumour necrosis factor receptors （sTNFR） on the structural and functional recovery in spinal cord transected rats. Methods GDNF-adenovirus and sTNFR-adenovirus were used to infect neural stem cells （NSCs） . The transgene NSCs were transplanted into the transected spinal cords in rats. The locomotor function of the rats was evaluated by BBB （Basso, Beattieand, Bresnahan） score and their histology by H＆E staining. Results High level GDNF and sTNFR proteins were found expressed in the spinal cords with transplanted neural stem cells which had been infected by recombinant GDNF-adenovirus and sTNFR- adenovirus. The BBB scores in group A were significantly higher than in group C. The BBB scores in groups A and C were remarkably higher than in group D. Conclusions Implantation of the neural stem cells infected with GDNF-adenovirus and sTNFR-adenovirus into spinal cords can alleviate the secondary pathogenic damage and promote the recovery of locomotor function in rats with spinal cord injury.