目的探讨ChABC、GDNF及Nogo-A抗体联合应用对大鼠脊髓完全性横断伤后脊髓再生修复及功能恢复的影响。方法采用大鼠胸段(T7-8)脊髓完全横切损伤模型,将SD大鼠随机分为6组:正常对照组、单纯横断组、ChABC(A组)、GDNF(G组)、Nogo-A抗体(N组)、ChABC+GDNF+Nogo-A抗体(AGN组)。于24周取材,行BDA示踪、NF-200、GAP-43、GFAP免疫组化染色。结果 A组、G组、N组BDA示踪染色,光镜下可见损伤区近侧端蓝色的示踪颗粒较多,损伤区很少有蓝色的再生神经纤维;AGN组可见蓝色的神经纤维通过损伤区并出现在损伤区的远侧段,损伤区中央为空泡样变,周围可见蓝染的纤维。NF-200免疫组化染色可见,A组、AGN治疗组NF阳性染色强于正常对照组及单纯横断组(P〈0.05),AGN组较G组、N组显著增强(P〈0.05)。A组、AGN治疗组GAP-43阳性染色强于正常对照组及单纯横断组(P〈0.05),AGN组较A、G、N组显著增强(P〈0.05)。正常对照组及单纯横断组GFAP阳性染色强于各治疗组(P〈0.05),A组、G组、N组与AGN组比较未见统计学差异(P〉0.05)。仅正常对照及AGN组可记录到SEP,但AGN组较对照组SEP潜伏期有所延长。结论 ChABC、GDNF、抗Nogo-A抗体单独或联合应用,能改善脊髓损伤区及两段的神经细胞功能,联合应用对脊髓再生修复优于各单一治疗组。
Objective To investigate regeneration and repair effect after ChABC,GDNF and Nogo-A Ab combination treatment for experimental spinal cord injury model.Methods Rat (T7-8 )complete spinal cord injury crosscutting animal model was established.The SD rats were randomly divided into 6 groups:normal group, sham operation group,simple transection group,A (ChABC)group,G (GDNF)group,N (Nogo-A antibody) group,and AGN (ChABC+GDNF+Nogo-A antibody)group.At 24 w after spinal cord injury,BDA tracer,NF-200,GAP-43,and GFAP immunohistochemistry were evaluated.Results BDA tracer of A group,G group and N group showed dye light,the proximal end of damaged zone showed the blue tracer particles,while damaged zone showed few blue regenerated nerve fibers.AGN group showed visible blue nerve fibers through the damaged zone and the distal segment in the damaged zone;the central zone of injury vacuolar degeneration showed the blue dyed fibers.NF-200 immunohistochemical staining showed NF-positive staining in A group,AGN was stronger than that in control group and simple transection group (P 〈0.05),AGN group than G group,N group significantly increased (P 〈0.05).GAP-43 positive staining in A group and AGN treatment group was stronger than in control group and simple transection group (P 〈0.05),AGN group significantly increased compared with A,G and N groups (P 〈0.05).GFAP positive staining in control group and simple transection group was stronger than in each treatment group (P 〈0.05),but A group,G group,N group and AGN group showed no significant differences (P 〉 0.05 ).SEP wave was detected in control group and AGN group,while the latency time was longer in AGN group than in control group.Conclusion ChABC,GDNF,and anti-Nogo-A antibody used alone or in combination can improve spinal cord injury and nerve cell function,and the joint application could improve regeneration after spinal cord injury than any monotherapy.