ChABC联合ADSC促进脱细胞同种异体神经支架移植后轴突再生的作用

Effects of chondroitinase ABC and adipose-derived stem cell transplantation on axon regeneration after acellular nerve allografting in rats

[目的]探讨硫酸软骨素酶ABC(ChABC)与脂肪源性干细胞(ADSC)联合应用对大鼠脱细胞同种异体神经支架移植后轴突再生的影响。[方法]36只成年大鼠随机分为脱细胞神经支架(ARSN)组、ChABC组、ADSC组、ChABC+ADSC组;坐骨神经功能指数(SFI)、电生理和胫前肌湿重比方法检测神经再生和运动功能的恢复;荧光显微镜观察神经移植体内PKH-26标记的ADSC;电镜检测神经移植体中有髓神经纤维数目、轴突直径和髓鞘厚度;免疫荧光法检测神经移植体中S100和GFAP蛋白表达。[结果]ChABC+ADSC组神经移植体内PKH-26标记的ADSC高于ADSC组(P<0.05)。与ARSN组比较,ChABC组、ADSC组和ChABC+ADSC组SFI增高,神经传导速度增高,潜伏时缩短,动作电位波幅增大,胫前肌湿重比率增高,有髓神经纤维数目、髓鞘厚度、轴突直径、S100和GFAP蛋白表达显著增加(P<0.05)。其中ChABC+ADSC组作用显著优于单独治疗组(P<0.05)。[结论]ChABC联合ADSC移植对大鼠脱细胞异体神经支架修复坐骨神经缺损后轴突再生的作用优于单独治疗组。

[Objective]To study the effect of adipose-derived stem cell（ADSC） transplantation and chondroitinase ABC（ChABC） treatment on axon regeneration after acellular nerve allograft repair of a sciatic nerve gap in rats.[Method]A total of 36 healthy Sprague-Dawley rats were divided randomly into acellular rat sciatic nerve（ARSN）,ChABC,ADSC,and ChABC ＋ ADSC groups.Immunofluorescence was used to detect PKH-26-labeled ADSCs in the nerve graft.Nerve regeneration and motor functional recovery were evaluated on the basis of the sciatic functions index（SFI）,electrophysiology,and the change in the tibialis anterior muscle wet weight.The mean myelinated fiber number,myelin sheath thickness,and axon diameter in the regenerated nerve were evaluated using toluidine blue staining.The expression levels of the S100 and GFAP proteins in the regenerated nerve were analyzed by immunofluorescence staining.[Result]The number of PKH-26 labeled ADSCs was higher in the ChABC ＋ ADSC group than in the ADSC group（P 0.05）.The SFI,nerve conduction velocity,wave amplitude,change in the tibialis anterior wet muscle weight,number of myelinated fibers,myelin sheath thickness,axon diameter,and S100 and GFAP protein levels were significantly higher in the ChABC,ADSC,and ChABC ＋ ADSC groups than in the ARSN group（P 0.05）,however,all 3 treatments significantly decreased the incubation period（P 0.05）.The ChABC ＋ ADSC group exhibited the strongest effect among the treatment groups（P 0.05）.[Conclusion]The combination of ADSC transplantation and ChABC treatment is significantly more effective than ARSN treatment alone at promoting axon regeneration after acellular nerve allograft repair of a sciatic nerve gap in rats.