不同断面神经束组吻合对周围神经再生影响的实验研究

目的 观察不同断面神经束组吻合对大鼠坐骨神经再生的影响。方法 将32只大鼠随机分为4组,各组大鼠均切断双侧坐骨神经后立即在显微镜下左侧施行不同断面神经束组吻合,右侧施行同一断面神经束组吻合。分别于第,2,4,6,8周检测两侧坐骨神经运动诱发电位的潜伏期和波幅,有髓神经纤维数。结果 各指标经统计学处理,术后第2周,实验侧和对照侧之间差异无统计学意义。结论 不同断面神经束组吻合在神经恢复后期有加速神经再生作用,优于同一断面神经束组吻合。     

应用人工神经修复周围神经损伤的研究

借助人工神经-明胶导管作载体,连接大鼠坐骨神经10mm缺损,观察周围神经再生的规律.结果表明,热脱水架桥处理的明胶导管组1个月后,其内壁已形成毛细血管网,2个月后导管被分解吸收,缺损部位神经再生完成.而用戊二醛交联组未见神经再生.手术后2个月和4个月进行锇染色、Bodian染色及S-100蛋白染色,2个月时,发现再生轴索已到达缺损部位的远端,新生髓鞘已经形成.结合麦芽凝集素辣根过氧化物酶(WGA-HRP)追踪显示,脊髓前角的运动神经元和后角的感觉神经末梢四甲基联苯胺显色法(TMB)反应呈阳性.机能检测可见诱发肌电图和大脑体性感觉诱发电位恢复.以上结果提示明胶是用来修复周围神经损伤的良好医用载体材料.     

大鼠坐骨神经盐酸氮芥作用后恢复的实验研究

观察大鼠坐骨神经节段(10 mm)经盐酸氮芥作用后其功能恢复情况.从而实现灭活侵犯神经干的肿瘤细胞的同时最大限度地保留周围神经的再生能力,进而探索出一理想的肢体恶性肿瘤的治疗方法.选26只健康雌性Wistar大白鼠,随机分成A组(盐酸氮芥处理组)13只、B组(NaCl对照组)13只.按30 mg/kg体重给予戊巴比妥钠腹腔麻醉后,显露右侧坐骨神经,A组取梨状肌下缘下一段长10 mm的神经干注入10%盐酸氮芥溶液0.1 mL,作用5 min.B组用0.9% NaCl处理神经做对照.盐酸氮芥作用后的大鼠坐骨神经有变性,其功能受损,经10 周恢复后从坐骨神经功能指数(SFI),电生理检查混合神经传导速度、神经动作电位波幅、肌肉诱发动作电位波幅以及光镜下行单位面积有髓神经纤维计数分析等指标评价显示A、B两组之间差异无显著性(P>0.05).在实验条件下,大鼠坐骨神经经盐酸氮芥作用后尚能保存神经再生修复的能力.     

局部非冻结性冷暴露对豚鼠周围神经形态结构的影响

在３６ 只成年雄性豚鼠上观察了动物后半身受到４ ±２ ℃局部冷暴露后周围神经形态结构的变化。结果表明：冷暴露后坐骨神经内的微血管充血;有髓纤维从施兰氏切迹和郎飞氏结附近开始呈现进行性的髓鞘变性,以暴露后３ 天最为严重。此时电镜显示髓鞘板层分离、断裂,间距明显增大。冷暴露后７ 天显示纤维再生与变性并存。与细纤维相比,粗纤维较早发生变性,也更为明显。在非冷暴露部位的上肢桡神经上未观察到上述变化     

Q&A

Q:为什么运动员和一般人的神经信息传输时间不同?(读者:杨涛)A:神经元是神经系统的基本结构和功能单位,它具有感受刺激、产生和传导兴奋的功能。兴奋的传递过程,包括在神经纤维上的传导和在神经元之间的传递。神经纤维的直径、髓鞘的厚度、结间段的长度和温度的高低是决定神经冲动(电信号)在有髓神经纤维上传导快慢的几个主要参数。直径粗、髓鞘厚、结间段长的纤维传导速度快。直径粗     

兔坐骨神经电损伤后背根神经节离子通道SCN9A、KV1．5表达变化的研究

电压依赖性离子通道是神经元和其他可兴奋细胞赖以产生可传导电信号的结构基础.实验观察兔坐骨神经在不同损伤电压情况下,背根神经节(DRG)上离子通道的表达变化.采用100只健康成年中国家兔分为阴性对照组、50 V、110 V和220 V 等4个电损伤组,取标本时间分为立即组、4周组.用免疫组织化学方法半定量分析DRG上钠钾离子通道的表达变化.结果电损伤后立即与4周后背根神经节(DRG)上离子通道的表达降低,且与电压呈负相关,220 V 4周时无明显恢复.说明离子通道在神经电损伤后的溃变程度和再生恢复取决于电压和时间.     

纤维环穿刺髓核抽吸法建立兔椎间盘退变模型

为探讨纤维环穿刺髓核抽吸法建立兔椎间盘退变模型的可行性。采用选取10月龄健康新西兰大白兔8只,雌雄不限,随机平均分为正常组和手术组。正常组不做处理,手术组术前于麻醉下行X线片,术中暴露椎间盘(L3-4、L4-5、L5-6),16G针纤维环穿刺抽吸髓核,术后4、8周X线片检查后收集椎间盘标本,大体观察、HE染色、甲苯胺蓝染色、番红"O"染色观察椎间盘退变。结果显示,X线片显示正常组椎间隙无明显改变,手术组4周、8周椎间隙狭窄,椎体前缘终板处变尖、毛糙,终板钙化,骨赘形成。手术组与正常组%DHI相比,差异具有明显统计学意义(P0.05)。组织病理学染色:正常组椎间盘结构完整,髓核与纤维环界限明显,排列整齐;术后4周髓核细胞明显减少,髓核与纤维环组织界限欠清晰,纤维环层状排列不整齐,术后8周髓核被纤维软骨组织替代,蛋白多糖含量明显减少。由此可知,纤维环穿刺髓核抽吸法成功的建立了兔椎间盘退变模型。     

芬必得对大鼠腰椎间盘突出症模型的疗效评价

目的 研究环氧化酶抑制剂芬必得对腰椎间盘突出症的作用。方法 通过压迫背根神经节及移植自体尾椎髓核至腰椎对大鼠进行造模。COX(cyclooxygenase)抑制剂芬必得给药剂量为0.051 g/kg体质量,连续28 d。对动物进行运动功能评分、机械刺激回缩反应测定、动物大腿周径测量及步态测量。给药28 d后取腰椎(L4～L6)及其神经用4%甲醛溶液固定后进行病理组织学检查,并取血清及神经根病变组织进行炎性因子浓度测定。结果 COX抑制剂芬必得能够改善模型动物的动物运动功能评分、50%机械性缩足反射阈值、大腿周径比(造模侧/对侧)、坐骨神经功能指数(SFI),背根神经节及椎间盘组织病理改变,降低血清和病变位置的炎症因子(PGE2、COX-2、NO、IL-1β、IL-6)的水平。结论 COX抑制剂可改善大鼠腰椎间盘突出症模型症状。     

胰岛素样生长因子-2对大鼠坐骨神经横断性损伤后再生修复的影响

目的 探讨胰岛素样生长因子-2(IGF-2)对大鼠坐骨神经横断性损伤后再生修复的影响.方法 构建大鼠右侧坐骨神经横断损伤模型,并行手术修复损伤.将40只Wistar雄性成年大鼠随机分为空白组、生理盐水组、IGF-2组、地塞米松组,术后2、4、6、8、12周分别采用足印实验检测坐骨神经功能指数;术后12周进行大鼠手术侧肌肉湿质量比检测,截取缝合端坐骨神经,进行HE染色及组织形态学观察.结果 术后IGF-2组SFI值明显高于空白组和生理盐水组(P<0.05);术后2、4、6周,3个测定时间点IGF-2组SFI值低于地塞米松组(P<0.05),8、12周,两个测定时间点IGF-2组与地塞米松组SFI值差异无统计学意义(P>0.05).IGF-2组较空白组与生理盐水组肌湿质量比明显增高(P<0.05),IGF-2组与地塞米松组肌湿质量比差异无统计学意义(P>0.05).HE染色显示:IGF-2组较空白组及生理盐水组损伤修复后瘢痕组织少,神经鞘修复良好.结论 IGF-2对于大鼠坐骨神经横断性损伤后的再生与修复有一定的促进作用.     

大鼠面神经颅外段的解剖及组织学研究

目的研究大鼠面神经的组成、走行、分支,为实验研究提供依据。方法显露面神经及各分支,测量各分支长度及外径,改良特殊三色法染色观察光镜结构。结果大鼠面神经出茎乳孔后前行,贴咬肌的表面分为颞支、颧支、颊支、下颌缘支和颈支至面部表情肌。面神经分支的长度和和中点外径依次为颞支(10.59 mm±0.34 mm,0.56±0.12 mm),颧支(15.48 mm±1.86 mm,0.70 mm±0.08 mm),颊支(26.18 mm±1.37 mm,1.01 mm±0.28 mm),下颌缘支(26.31 mm±1.32 mm,0.91 mm±0.20 mm),颈支(17.54 mm±0.75 mm,0.64 mm±0.15 mm)。特殊三色法染色光镜下可见神经内膜呈绿色,完整;神经束较少,多为2~3束。大量神经纤维整齐排列,轴突位于中央,较小,呈蓝绿色,髓鞘呈枣红色,红细胞呈鲜红色,细胞核呈紫蓝色。结论应选择大鼠面神经颊支和下颌缘支进行缺损修复模型的制备。     

Nerve regeneration factor promotes nerve regeneration in rat

Nerve regeneration factor (NRF) extracted from an oral liquid of traditional Chinese medicine, as a nerve growth decoction has been reported by previous studies to exert effects of promoting nerve growth and preventing neuron apoptosis. For new insights into the function of NRF on primary cultured neurons, we investigated the neurite outgrowth in cultured rat dorsal root ganglion (DRG) explant treated with NRF by immunofluorescence and the gene and protein expressions of neurofilament-H(NF-H) and growth associated protein 43 (GAP43) in the cultured rat DRG neurons by real-time quantitative RT-PCR and Western blotting, respectively. In addition, we used a rat model of sciatic nerve crush to evaluate the effect of NRF on regeneration of injured sciatic nerve by a combination of walk track analysis, electrophysiological and histological assessments. The in vitro experiments indicated that NRF promoted the neurite growth of DRGs and the expression of NF-H and GAP43 at mRNA and protein levels in DRG neurons, and in vivo experiments showed that NRF improved peripheral nerve regeneration and functional recovery.     

眼镜蛇毒神经生长因子对成年猫坐骨神经损伤治疗效果的实验室观察

目的 :探讨从眼镜蛇毒分离纯化出的神经生长因子(nervegrowthfactor,NGF)对成年猫坐骨经损伤后的影响。方法 :本实验制成成年猫坐骨神经损伤模型 ,损伤局部注射蛇毒NGF(2μg/kg/d) ,分别治疗10d和30d ,并与对照组(损伤坐骨神经 ,不给药物)比较。结果 :眼镜蛇毒NGF在神经损伤早期应用能减轻神经纤维发生的溃变 ,促进神经纤维再生。结论 :损伤局部长时间注射NGF会导致神经纤维增生过度 ,丧失传导功能     

bFGF对自体神经移植后神经再生和神经元相关性的影响

目的探讨bFGF对自体神经移植后神经再生和神经元相关性的影响。方法切断大白鼠右侧坐骨神经10mm,在原位作神经外膜缝合,实验组注射bFGF100u/d,共10d,对照组注射生理盐水10d,术后12周取材。光镜观察,并计数再生神经纤维和神经元的绝对值,得出恢复率以及用统计学方法得出它们的相关性。结果两组均可见再生神经纤维通过吻合口,并向远端延伸。两组神经再生率与脊髓前角运动细胞或者脊神经节细胞存活率的相关系数均非常接近1,但实验组和对照组的组间相关系数无显著性差异。结论周围神经再生率与相应的脊髓前角运动细胞或者脊神经节细胞存活率之间的正相关关系是固有的,不会随bFGF的使用而改变     

哺乳动物神经纤维的双向脉冲选择性刺激研究

为了研究神经纤维的双向脉冲选择性刺激方法,对神经纤维的各种物理模型进行了研究和比较,建立了哺乳动物有髓神经纤维的仿真系统,并利用该系统对以前工作中提出的３种双向脉冲的选择性刺激方法（单电极,双电极和三电极方法）进行了研究,发现也可以实现复合神经干中细神经纤维的选择性兴奋,证明这些方法同样适用于哺乳动物的神经纤维模型。     

大鼠坐骨神经切断后经皮低频高强度电刺激相应脊髓节段中GAP-43表达的变化

目的探讨成年Wister大鼠在坐骨神经切断后GAP-43于相应脊髓节段前角运动神经元内的表达变化.方法选取健康成年雄性Wister大鼠60只,将坐骨神经切断,随机分为实验组和对照组,实验组给予经皮低频高强度电刺激,分别于术后1,2,4,8,12,16周处死,取其L4~L6脊髓,利用免疫组织化学技术检测GAP-43在相应脊髓节段中的表达变化,并利用影像分析系统进行统计学分析.结果对照组:1周时前角细胞胞体内GAP-43有明显表达,4周时达到高峰,5~8周时逐渐下调,9~16周时GAP-43在前角细胞胞体内中仍有少量表达,并呈弱阳性.实验组:1周时前角细胞胞体内GAP-43有明显表达,2周时达到高峰,且在4~16周时在神经元中仍有表达,并呈阳性.结论坐骨神经切断可导致成年大鼠相应脊髓节段中前角运动神经元GAP-43表达明显增加,可证明在周围神经损伤后神经元的再生能力增强,但时效很短.而给予低频高强度电刺激疗法后,GAP-43的表达在时长和量上都有明显增加.     

Biocompatibility studies of silk fibroin-based artificial nerve grafts in vitro and in vivo

Silk fibroin(SF)has been used extensively in the biomedical field including tissue engineering for the generation of artificial bones,skins or ligaments.We have previously reported on good in vitro biocompatibility of SF fibers with peripheral nerve tissues and cells.In the present study,we developed a novel design of the SF-based artificial nerve graft(SF graft)which was composed of a SF-nerve guidance conduit(NGC)inserted with SF fibers.MTT assay was performed to determine the cytotoxicity of the SF-NGC extract fluid on the cultured L929 cells derived from an immortalized mouse fibroblast cell line.In addition,this SF graft was implanted into adult rats for bridging a 10-mm long sciatic nerve defect.The following-up experiments at initial stage(1-4 week)of nerve regeneration including routine blood tests and histochemical investigation were conducted to evaluate the in vivo biocompatibility of the SF graft with peripheral nerves.The results demonstrated that the SF-NGC graft was biocompatible with the surrounding tissues and cells due to its low inflammatory potential with a grade 0 under the U.S.Pharmacopeia guidelines and it was generally suitable to a certain degree for bridging peripheral nerve defects in virtue of supporting Schwann cell adherence,expansion and migration.Therefore the SF graft is a promising alternative to classical autografts for peripheral nerve repair.     

Dog sciatic nerve gap repaired by artificial tissue nerve graft

The feasibility of repairing dog sciatic nerve damage by using a biodegradable artificial tissue nerve graft enriched with neuroregenerating factors is investigated. The artificial nerve graft was implanted to a 30 mm gap of the sciatic nerve damage in 7 dogs. The dogs with the same nerve damage that were repaired by interposition of the autologous nerve or were given no treatment served as control group 1 or 2, respectively. The observations include gross and morphological observations, immune reaction, electrophysiological examination, fluorescence tracing of the neuron formation and the number of the neurons at the experimental sites, etc. Results showed that 6 months after the implantation of the graft, the regenerated nerve repaired the damage of the sciatic nerve without occurrence of rejection and obvious inflammatory reaction in all 7 dogs, and the function of the sciatic nerve recovered with the nerve conduction velocity of (23.91±11.35)m/s. The regenerated neurons and the forming of axon could be observed under an electron microscope. This proves that artificial tissue nerve graft transplantation can bridge the damaged nerve ends and promote the nerve regeneration.     

Functional regeneration of sensory axons into the adult spinal cord

The arrest of dorsal root axonal regeneration at the transitional zone between the peripheral and central nervous system has been repeatedly described since the early twentieth century. Here we show that, with trophic support to damaged sensory axons, this regenerative barrier is surmountable. In adult rats with injured dorsal roots, treatment with nerve growth factor (NGF), neurotrophin-3 (NT3) and glial-cell-line-derived neurotrophic factor (GDNF), but not brain-derived neurotrophic factor (BDNF), resulted in selective regrowth of damaged axons across the dorsal root entry zone and into the spinal cord. Dorsal horn neurons were found to be synaptically driven by peripheral nerve stimulation in rats treated with NGF, NT3 and GDNF, demonstrating functional reconnection. In behavioural studies, rats treated with NGF and GDNF recovered sensitivity to noxious heat and pressure. The observed effects of neurotrophic factors corresponded to their known actions on distinct subpopulations of sensory neurons. Neurotrophic factor treatment may thus serve as a viable treatment in promoting recovery from root avulsion injuries. I     

前列腺素E1,锌离子对大鼠感觉神经元的保护作用

目的探讨环化酶激活剂(AdenyIate cyclase activator)对周围神经损伤后感觉神经元存活和变性的影响.方法大鼠坐骨神经夹毁模型、分组,术后每d分别局部注射一定剂量的PGE1,hβNGF,共14 d.另一组从术前10 d开始喂以高锌饲料各组术后21 d取背根节(DRG)制片染色,用图像分析法观测PGE1和锌对DRG细胞数、核偏心率和核等圆径的影响,以人胎盘神经生长因子(hβNGF)作阳性对照.结果锌显著降低感觉神经元的死亡率,显著减轻核偏移、抑制核等圆径增大,与NGF组相比,P＞0 05.PGE1也明显减少神经元死亡,与夹毁组相比,P＜0 05结论神经损伤后锌促进感觉神经元存活和减轻细胞变性的保护作用与NGF无差别.PGE1对感觉神经元的保护作用不及NGF和锌.     

人发角蛋白桥接周围神经缺损的功能评价

目的通过在神经再生室内植入人发角蛋白,观察神经的功能恢复情况,为桥接周围神经缺损寻求新的替代材料。方法将18只新西兰兔的左侧坐骨神经切断,造成lOmm缺损,实验组用人发角蛋白桥接,对照组用空硅胶管桥接,术后检测坐骨神经功能指数、电生理、腓肠肌湿重,观察神经功能的恢复情况。结果8周时,试验组坐骨神经功能指数值优于对照组（P〈0．01）;12周时,试验组坐骨神经功能指数、潜伏期和神经传导速度及波幅明显好于对照组（P〈0．01）。结论人发角蛋白能促进兔坐骨神经功能的恢复是桥接周围神经缺损的理想材料。