Netrin-1、Slit2在夹脊电针结合神经松动术调节兔坐骨神经损伤后Rho GTPases失衡的作用

Role of Netrin-1 and Slit2 in regulating the imbalance of Rho GTPases after Jiaji electroacupuncture combined with nerve mobilization for sciatic nerve injury in rabbits

目的 观察夹脊电针结合神经松动术疗法对兔坐骨神经损伤后神经传导速度及Netrin-1、Slit2、Rho GTPases表达的影响。方法 将216只新西兰家兔随机分为正常对照组、病毒空载组、夹脊电针结合神经松动术组、Netrin-1组、Slit2组、Netrin-1+Slit2腺病毒结合组,按术后1、2、4周三个时间点分为3个亚组,每个亚组12只。钳夹法造兔左侧坐骨神经SunderlandⅢ度损伤模型。正常对照组无干预;各病毒组在模型制备时完成目的基因转染病毒注射;夹脊电针结合神经松动术组术后3 d即进行夹脊电针及神经松动术治疗。肌电图测量兔坐骨神经传导速度,取损伤坐骨神经和L4-6脊髓,荧光实时定量聚合酶链反应检测Netrin-1 mRNA、Slit2mRNA表达,Western blotting检测Rac1、Cdc42、RhoA蛋白表达。结果 术后1、2、4周,夹脊电针结合神经松动术组、Netrin-1+Slit2腺病毒结合组坐骨神经神经传导速度高于Netrin-1组和Slit2组(P <0.05);电针结合神经松动术组、 Netrin-1+Slit2腺病毒结合组Netrin-1和Slit2mRNA表达高于正常对照组、病毒空载组(P <0.05);电针结合神经松动术组和Netrin-1+Slit2腺病毒结合组Rac1、Cdc42蛋白表达高于Netrin-1组、Slit2组和病毒空载组(P <0.05),而RhoA蛋白表达低于Netrin-1组、Slit2组和病毒空载组(P <0.05)。结论 夹脊电针结合神经松动术促进轴突再生可能是通过调节Netrin-1、Slit2的表达,恢复Rho GTPases酶系统失衡状态,重组细胞骨架,促进周围神经损伤后的再生。

Objective To observe the effect of Jiaji electroacupuncture combined with neurodynamic mobilization on nerve conduc‐tion velocity and the expression of Netrin-1,Slit2 and Rho GTPases after sciatic nerve injury in rabbits.Methods A total of 216 New Zealand rabbits were randomly divided into nomal control group(NC),vector virus group(VV),Jiaji electroacupuncture combined with neurodynamic mobilization group(EN),Netrin-1 group(N1),Slit2 group(S2)and N1+S2 group.Each group was divided into three subgroups according to the postoperative treatment time points(one,two and four weeks),with twelve rabbits in each subgroup.The rabbit model of Sun‐derlandⅢdegree injury of the left sciatic nerve was established by clamping method.There was no intervention in NC,and virus was injected during the preparation of the model.Jiaji electroacupuncture and neurodynamic mobilization were administrated three days after operation in EN group.Nerve conduction velocity of sciatic nerve was measured with electromyography.The sciatic nerve and L4-6 spinal cords were obtained,the expression of Netrin-1 and Slit2 mRNA were detected real-time quantitative polymerase chain reaction and immunofluores‐cence double staining,and the expression of Rac1,Cdc42 and RhoA protein were observed via Western blotting.Results One,two and four weeks after modeling,the nerve conduction velocity was more in EN group and N1+S2 group than in N1 group and S2 group(P<0.05);the Netrin-1 and Slit2 mRNA expression were higher in the EN group and N1+S2 group than in the VV group and NC group(P<0.05);the protein expression of Rac1 and Cdc42 were higher,and the protein expression of RhoA was lower in EN group and N1+S2 group than in N1 group,S2 group and VV group(P<0.05).Conclusion Jiaji electroacupuncture combined with nerve mobilization may promote the axonal regeneration by regulat‐ing the expression of Netrin1 and Slit2,adjusting the imbalance of Rho GTPases enzyme system,and then pro‐moting cytoskeleton reorganization and peripheral nerve regeneration after injury.