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  

Effect of short-term low-frequency electrical stimulation on nerve regeneration of delayed nerve defect during operation

作　　者：焦海山[1] 肖波[1] 王晓冬[2] 李东印[1] 宋悦宁[1] 郑辉[1] 刘晓梅[1] 

机构地区：[1]苏州卫生职业技术学院基础医学教研室,江苏苏州215009 [2]南通大学医学院组织学与胚胎学教研室,江苏南通226001

出　　处：《中国修复重建外科杂志》2017年第3期335-344,共10页Chinese Journal of Reparative and Reconstructive Surgery

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)~~

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(P>0.05)。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�Objective To explore the effect of short-term low-frequency electrical stimulation （SLES） during operation on nerve regeneration in delayed peripheral nerve injury with long gap. Methods Thirty female adult Sprague Dawley rats, weighing 160-180 g, were used to prepare 13-mm defect model by trimming the nerve stumps. Then all rats were randomly divided into 2 groups, 15 rats in each group. After nerve defect was bridged by the contralateral normal sciatic nerve, SLES was applied in the experimental group, but was not in the control group. The spinal cords and dorsal root ganglions （DRGs） were harvested to carry out immunofluorescence histochemistry double staining for growth-associated proteins 43 （GAP-43） and brain-derived neurotrophic factor （BDNF） at 1, 2, and 7 days after repair. Fluorogold （FG） retrograde tracing was performed at 3 months after repair. The mid-portion regenerated segments were harvested to perform Meyer＇s trichrome staining, immunofluorescence double staining forneurofilament （NF） and soluble protein 100 （S-100） on the transversely or longitudinal sections at 3 months atter repair. The segment of the distal sciatic nerve trunk was harvested for electron microscopy and morphometric analyses to measure the diameter of the myelinated axons, thickness of myelin sheaths, the G ratio, and the density of the myelinated nerve fibers. The gastrocnemius muscles of the operated sides were harvested to measure the relative wet weight ratios. Karnovsky-Root cholinesterase staining of the motor endplate was carried out. Results In the experimental group, the expressions of GAP-43 and BDNF were higher than those in the control group at 1 and 2 days after repair. The number of labeled neurons in the anterior horn of gray matter in the spinal cord and DRGs at the operated side from the experimental group was more than that from the control group. Meyer＇s trichrome staining, immunofluorescence double staining, and the electron microscopy observation showed that the regenerated n

关 键 词：短期低频电刺激 周围神经 陈旧性损伤 神经缺损修复 大鼠 

分 类 号：R651.3[医药卫生—外科学]