本文選題：短期低頻電刺激　切入點(diǎn)：周圍神經(jīng)　出處：《中國(guó)修復(fù)重建外科雜志》2017年03期

【摘要】：目的探討術(shù)中短期低頻電刺激(short-term low-frequency electrical stimulation,SLES)對(duì)陳舊性周圍神經(jīng)缺損再生能力的影響。方法成年雌性SD大鼠30只,體質(zhì)量160~180 g,制備大鼠坐骨神經(jīng)13 mm長(zhǎng)陳舊性缺損模型,隨機(jī)分為實(shí)驗(yàn)組和對(duì)照組,每組15只。實(shí)驗(yàn)組切取對(duì)側(cè)正常坐骨神經(jīng)橋接缺損并施以SLES,對(duì)照組同法修復(fù)神經(jīng)缺損但不予以電刺激。修復(fù)術(shù)后1、2、7 d取大鼠脊髓腰膨大段(L_(4、5))以及相應(yīng)背根神經(jīng)節(jié),行抗生長(zhǎng)相關(guān)蛋白43(growth-associated proteins 43,GAP-43)及抗腦源性神經(jīng)營(yíng)養(yǎng)因子(brain-derived neurotrophic factor,BDNF)雙重免疫熒光染色觀察。修復(fù)術(shù)后3個(gè)月行熒光金逆行示蹤實(shí)驗(yàn)。取再生神經(jīng)組織中段作橫、縱冰凍切片,分別行Meyer神經(jīng)三色染色和抗鼠神經(jīng)絲蛋白(neurofilament,NF)及可溶性蛋白100(soluble protein100,S-100)雙重免疫熒光染色觀察,并取術(shù)側(cè)遠(yuǎn)端神經(jīng)行透射電鏡觀察,測(cè)量有髓神經(jīng)纖維、軸突直徑及髓鞘厚度,計(jì)算有髓神經(jīng)纖維密度及G比值。取術(shù)側(cè)腓腸肌計(jì)算相對(duì)濕重比,并行Karnovsky-Root運(yùn)動(dòng)終板膽堿酯酶組織學(xué)染色觀察。結(jié)果實(shí)驗(yàn)組修復(fù)術(shù)后1、2 d,術(shù)側(cè)對(duì)應(yīng)運(yùn)動(dòng)神經(jīng)元和感覺神經(jīng)元GAP-43和BDNF表達(dá)上調(diào)快于對(duì)照組。熒光金逆行示蹤實(shí)驗(yàn)示,實(shí)驗(yàn)組術(shù)側(cè)脊髓前角及相應(yīng)背根神經(jīng)節(jié)中標(biāo)記的神經(jīng)元多于對(duì)照組。實(shí)驗(yàn)組再生神經(jīng)Meyer神經(jīng)三色染色和雙重免疫熒光染色均顯示再生神經(jīng)組織發(fā)育優(yōu)于對(duì)照組。透射電鏡觀察示再生神經(jīng)纖維成簇分布,實(shí)驗(yàn)組軸突直徑、髓鞘厚度、有髓神經(jīng)纖維密度及G比值均高于對(duì)照組,但差異無統(tǒng)計(jì)學(xué)意義(P0.05)。實(shí)驗(yàn)組術(shù)側(cè)腓腸肌相對(duì)濕重比高于對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(t=4.633,P=0.000)。腓腸肌Karnovsky-Root運(yùn)動(dòng)終板膽堿酯酶組織學(xué)染色示,實(shí)驗(yàn)組運(yùn)動(dòng)終板的形態(tài)和數(shù)量略優(yōu)于對(duì)照組。結(jié)論術(shù)中SLES能在一定程度上提高大鼠短期(1個(gè)月)長(zhǎng)距離陳舊性周圍神經(jīng)缺損的再生能力。
[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.
【作者單位】： 蘇州衛(wèi)生職業(yè)技術(shù)學(xué)院基礎(chǔ)醫(yī)學(xué)教研室;南通大學(xué)醫(yī)學(xué)院組織學(xué)與胚胎學(xué)教研室;
【基金】：蘇州市科技局資助項(xiàng)目(STSD2011074) 江蘇省衛(wèi)生廳衛(wèi)生職業(yè)技術(shù)教育研究室立項(xiàng)課題(J201301) 蘇州衛(wèi)生職業(yè)技術(shù)學(xué)院立項(xiàng)課題(szwzy200910) 江蘇省“青藍(lán)工程”資助項(xiàng)目(蘇教師[2012]39號(hào))~~
【分類號(hào)】：R651.3

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