硫酸镁联合自体激活雪旺细胞移植治疗脊髓损伤

Treatment of Spinal Cord Injury by Magnesium Sulfate Combined with Autologous Activated Schwann Cells Transplantation in Rats

目的:探讨腹腔注射硫酸镁(MgSO4)联合局部移植自体激活雪旺细胞(AASCs)治疗脊髓损伤的疗效。方法:通过结扎大鼠双后肢隐神经激活自体雪旺细胞(SCs),并进行体外分离、培养、传代获得大量细胞。Wistar大鼠48只以IMPACTOR MODEL-Ⅱ打击器制备脊髓胸10(T10)损伤模型。随机分为单纯DMEM移植空白对照组(A组)、MgSO4腹腔注射组(B组)、AASCs移植组(C组)和MgSO4腹腔注射联合AASCs移植组(D组)。对各组实验动物脊髓损伤后肢体功能的恢复情况进行行为学评分(BBB评分),生物素葡聚糖胺(BDA)示踪皮质脊髓束(corticospinal tract,CST),取出动物损伤段脊髓进行免疫荧光染色。结果:所有大鼠在脊髓损伤后出现下肢截瘫,各组BBB评分差异有统计学意义(P<0.05);C组和D组脊髓损伤区及其远端BDA标记的神经纤维数量明显多于其他组;各组脊髓NF200和GFAP免疫荧光染色阳性面积差异均有统计学意义(P<0.05),D组阳性面积明显高于其他实验组(P<0.05)。结论:MgSO4联合AASCs能保护脊髓损伤后神经功能,并促进其轴突再生。

Objective：To observe the effect of magnesium sulfate （MgSO4） intraperitoneally and transplantation of autologous activated schwann cells （AASCs） on the spinal cord injury in Wistar rats.Methods：The hibateral saphenous nerves of rats were ligated directly for 1 week to activate Schwann cells （SCs） and to culture them using the tissue clot method. The clone was passaged continuously for 4 generations and large number of clones was obtained.Then 48 female Wistar rats were randomly divided into 4 groups, DMEM group（n=12, group A）,MgSO4 group（n=12,group B）,SCs group（n=12, group C）,and combined transplantation group （n=12,group D） before transplantation. The ceils were purified to the injured T10 spinal cord of rats （IMPACTOR MODEL-Ⅱ type weight drop apparatus） before transplantation. The recoveries of the lower extremity were observed using Basso-Beattie-Bresnahan （BBB）locomotor scoring system.Twelve weeks later, two rats in each group selected at random were injected with 10% biotinylated dextran amine （BDA）in the double cerebra cortex for anterograde labeling of CST.After 2 weeks ,these rats were killed and perfused with 4% polyoxymethylene.The spinal cords including the segment of defect were taken out and cut into 5μm thick sections at 0 ℃.Immunohistochemistry was used to detect NF-200 and GFAP. Results： The BBB scores were higher in groups C and D than those in groups A and B. The regenerated nerve fiber stained by BDA was seen in the defected position in groups C and D, but almost nothing in the injured site in groups A and B.There was significant difference in NF-200 and GFAP between groups A, B, C and D, more in group D than those of other groups （P 〈 0.05）. Conclusion： Treatment of magnesium sulfate combined with autologous activate schwann cells transplantation can protect injured neuron, and promote regeneration of axon in spinal cord injury site.