术中短期低频电刺激对陈旧性周围神经缺损再生能力的影响

发布时间：2018-03-30 13:50

本文选题：短期低频电刺激　切入点：周围神经　出处：《中国修复重建外科杂志》2017年03期

【摘要】：目的探讨术中短期低频电刺激(short-term low-frequency electrical stimulation,SLES)对陈旧性周围神经缺损再生能力的影响。方法成年雌性SD大鼠30只,体质量160~180 g,制备大鼠坐骨神经13 mm长陈旧性缺损模型,随机分为实验组和对照组,每组15只。实验组切取对侧正常坐骨神经桥接缺损并施以SLES,对照组同法修复神经缺损但不予以电刺激。修复术后1、2、7 d取大鼠脊髓腰膨大段(L_(4、5))以及相应背根神经节,行抗生长相关蛋白43(growth-associated proteins 43,GAP-43)及抗脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)双重免疫荧光染色观察。修复术后3个月行荧光金逆行示踪实验。取再生神经组织中段作横、纵冰冻切片,分别行Meyer神经三色染色和抗鼠神经丝蛋白(neurofilament,NF)及可溶性蛋白100(soluble protein100,S-100)双重免疫荧光染色观察,并取术侧远端神经行透射电镜观察,测量有髓神经纤维、轴突直径及髓鞘厚度,计算有髓神经纤维密度及G比值。取术侧腓肠肌计算相对湿重比,并行Karnovsky-Root运动终板胆碱酯酶组织学染色观察。结果实验组修复术后1、2 d,术侧对应运动神经元和感觉神经元GAP-43和BDNF表达上调快于对照组。荧光金逆行示踪实验示,实验组术侧脊髓前角及相应背根神经节中标记的神经元多于对照组。实验组再生神经Meyer神经三色染色和双重免疫荧光染色均显示再生神经组织发育优于对照组。透射电镜观察示再生神经纤维成簇分布,实验组轴突直径、髓鞘厚度、有髓神经纤维密度及G比值均高于对照组,但差异无统计学意义(P0.05)。实验组术侧腓肠肌相对湿重比高于对照组,差异有统计学意义(t=4.633,P=0.000)。腓肠肌Karnovsky-Root运动终板胆碱酯酶组织学染色示,实验组运动终板的形态和数量略优于对照组。结论术中SLES能在一定程度上提高大鼠短期(1个月)长距离陈旧性周围神经缺损的再生能力。
[Abstract]:Objective to investigate the effect of short-term low-frequency electrical stimulation (SLES) on the regeneration ability of old peripheral nerve defects in rats. Methods Thirty adult female SD rats with body weight of 160 ~ 180 g were used to establish the rat model of sciatic nerve 13 mm long old defect. They were randomly divided into experimental group and control group. 15 rats in each group were removed from the contralateral normal sciatic nerve graft defect in the experimental group and treated with SLES. In the control group, the nerve defect was repaired with the same method without electrical stimulation. The anti-growth related protein (43(growth-associated proteins 43) and the anti-brain-derived neurotrophic factor (BDNF) were observed by double immunofluorescence staining. The fluorescent gold retrograde tracing test was performed 3 months after repair. The middle segment of regenerative nerve tissue was taken as transverse and longitudinal frozen sections. Meyer nerve trichrome staining, anti-mouse neurofilamentus (NFN) and soluble protein 100(soluble protein 100S-100) were observed by double immunofluorescence staining. The distal nerve of the operation was observed by transmission electron microscopy. The myelinated nerve fibers, axons diameter and myelin sheath thickness were measured. The density and G ratio of myelinated nerve fibers were calculated. The relative wet weight ratio of gastrocnemius muscle was calculated. Results the expression of GAP-43 and BDNF in the corresponding motor neurons and sensory neurons in the experimental group was increased more rapidly than that in the control group at 1 and 2 days after repair. The fluorescence gold retrograde tracing test showed that the expression of GAP-43 and BDNF in the corresponding motor neurons and sensory neurons in the experimental group was higher than that in the control group. The number of labeled neurons in the anterior horn of spinal cord and the corresponding dorsal root ganglion in the experimental group was more than that in the control group, and the Meyer and double immunofluorescence staining of regenerated nerve in the experimental group showed that the tissue development of regenerated nerve was superior to that of the control group. Transmission electron microscopy (TEM) showed that regenerated nerve fibers were clustered. The diameter of axon, the thickness of myelin sheath, the density of myelinated nerve fiber and G ratio in experimental group were higher than those in control group, but the difference was not significant (P 0.05). The relative wet weight ratio of gastrocnemius muscle in experimental group was higher than that in control group. The difference was statistically significant. The Karnovsky-Root motor end-plate of gastrocnemius was stained with cholinesterase. Conclusion intraoperative SLES can improve the regeneration ability of long distance old peripheral nerve defects in rats in a certain extent.
【作者单位】：苏州卫生职业技术学院基础医学教研室;南通大学医学院组织学与胚胎学教研室;
【基金】：苏州市科技局资助项目(STSD2011074) 江苏省卫生厅卫生职业技术教育研究室立项课题(J201301) 苏州卫生职业技术学院立项课题(szwzy200910) 江苏省“青蓝工程”资助项目(苏教师[2012]39号)~~
【分类号】：R651.3

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