嗅粘膜源性嗅鞘细胞移植联合NGF对脊髓损伤的修复

Repairs to the spinal cord by transplantation of olfactory ensheathing cells from the olfactory mucosa and NGF in adult rats

目的:探讨大鼠嗅粘膜嗅鞘细胞(olfactory ensheathing cells,OECs)移植联合神经生长因子(nervegrowth factor,NGF)修复脊髓急性损伤的可行性和疗效,并观察二者的协同作用。方法:自成年Wistar大鼠嗅粘膜取材,采用差速贴壁和阿糖胞苷抑制纯化相结合的方法培养纯化嗅鞘细胞。建立大鼠脊髓半切模型,并随机分成对照组(A组),OECs组(B组)和OECs+NGF组(C组),在不同时段进行肢体活动的BBB评分。8周后取脊髓标本进行组织学检查,评价对脊髓损伤的修复作用。结果:B组和C组2周后BBB评分高于A组(P<0.05),C组自4周后明显优于A组和B组(P<0.01)。组织学检查表明,C组可明显促进轴突再生。结论:OECs联合NGF具有良好的脊髓损伤修复作用,且二者具有明显的协同作用。

Objective： To investigate the effect of transplantation of olfactory ensheathing cells （OECs） originating from the olfactory mucosa and nerve growth factor（NGF） to repair spinal cords. Methods： The olfactory mucosa was separated from the nasal septum of adult Wistar rats. The OECs were purified by different attachment time with other types of cells and Ara-c inhibition, and the nerve growth factor receptor p75 （p75 NGFR） immunohistochemistry. The spinal cord samples were hemisected and they were randomly divided into three groups： the control group （group A）, the olfactory ensheathing cell group（group B）, and the olfactory ensheathing cells and the nerve growth factor group（group C）. The Basso, Beattie, and Bresnahan locomotor rating scale（BBB） was observed at 2, 4, 6 and 8 weeks. Histological changes, the axon regeneration and the survival of OECs were determined 8 weeks later by an electron microscope, hematoxylin and eesin（HE） and immunohistochemistry staining. Result： The BBB grade of groups B and C was significantly higher than that of group A at 2 weeks, and that of group C was significantly higher than that of groups A and B. Histology revealed that the regeneration of axons was significantly promoted in group C. Conclusion： Transplantation of OECs and NGF is great for spinal cord repairs, and the two have a significant synergistic effect.