神经生长因子对大鼠视觉诱发电位的影响

目的和方法：本研究主要通过电生理方法,测定经CS2所致视神经损害的大鼠在治疗前后视觉诱发电位的变化,以证实神经生长因子对受损神经的治疗效果。结果：大鼠视神经损伤模型经神经生长因子治疗20d后,模式反转诱发电位和闪光诱发电位的潜伏期与对照组相比均有明显的缩短,并有量－效关系。结论：神经生长因子能明显改善视神经传导功能,提示神经生长因子对视神经损伤有一定的治疗作用。     

泼尼松灌胃与肌内注射对大鼠骨密度、骨生物力学性能及骨代谢的影响

目的观察泼尼松灌胃与肌内注射两种不同给药方法对大鼠骨密度、骨生物力学及骨代谢的影响。方法将45只SPF级雄性SD大鼠随机分为3组（正常组15只、灌胃组15只、肌内注射组15只）,其中正常组大鼠作为阴性对照,予0.9%生理盐水灌胃2 m L/d;灌胃组大鼠给予泼尼松0.5 mg/（kg·d）灌胃;肌内注射组大鼠给予泼尼松0.5 mg/（kg·d）;12周后测定离体的大鼠椎体骨密度及血清β-CTX、PINP水平变化,采用三点弯曲试验测量股骨皮质骨最大载荷、弹性载荷、断裂载荷等生物力学指标。结果与正常组相比,灌胃组及肌内注射组大鼠椎骨骨密度值均显著性降低（P〈0.05）;与灌胃组相比,肌内注射组大鼠椎骨骨密度显著下降（P〈0.05）;与正常组相比,灌胃组及肌内注射组大鼠股骨的弹性载荷、最大载荷、断裂载荷均显著降低（P〈0.05）,肌内注射组与灌胃组大鼠的弹性载荷、最大载荷、断裂载荷相比差异无显著性（P〉0.05）。与正常组相比,灌胃组及肌内注射组大鼠中血清β-CTX水平均显著升高（P〈0.05）而PINP水平均显著降低（P〈0.05）,与灌胃组相比,肌内注射组大鼠血清β-CTX水平显著升高（P〈0.05）而PINP水平显著降低（P〈0.05）。骨组织切片HE染色显示：肌内注射组大鼠的骨小梁明显纤细疏松,造血组织明显减少,脂肪组织明显增多。结论泼尼松对大鼠的骨密度、骨生物力学及骨代谢指标都有影响,而肌内注射泼尼松比口服对骨密度、骨强度、骨代谢的影响更大,更易造成骨质疏松症。因此,建议临床使用泼尼松时选择口服作为给药方式更安全。     

加味四妙散对尿酸钠诱导急性痛风性关节炎大鼠的疗效机制研究

目的:研究急性痛风性关节炎大鼠应用加味四妙散的疗效,并研究其作用机制。方法:将60只雄性Wistar大鼠按照随机数字法随机分为5组,每组各12只,分组情况为:空白对照组、模型对照组、低剂量组(8.875 g/kg·d)、中剂量组(17.750 g/kg·d)及高剂量组(35.500 g/kg·d)。模型建立后每天灌胃治疗,持续3 d。计算并比较各组大鼠造模后4 h、24 h、48 h、72 h时踝关节的肿胀度。检测并比较各组大鼠血清白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)及尿酸(UA)水平。结果:(1)造模24h后,模型对照组大鼠关节肿胀度达到峰值,并且明显大于空白对照组,具有显著性差异(P0.05);模型对照组大鼠关节肿胀度明显大于各剂量组,同时高剂量组大鼠关节肿胀度明显小于中剂量组和低剂量组,均具有显著性差异(P0.05)。造模48 h后,各组大鼠关节肿胀度均继续下降,且各剂量组大鼠关节肿胀度明显小于模型对照组,均具有显著性差异(P0.05)。造模72 h后,各治疗组大鼠关节肿胀度均明显小于模型对照组,同时高剂量组大鼠关节肿胀度明显小于中剂量组和低剂量组,均具有显著性差异(P0.05)。(2)与空白对照组比较,模型对照组大鼠血清IL-1β、TNF-α及UA水平均明显偏高,具有显著性差异(P0.05);与模型对照组、低剂量组比较,中剂量组和高剂量组大鼠血清IL-1β、TNF-α及UA水平均明显偏低,具有显著性差异(P0.05)。结论:加味四妙散35.500 g/kg剂量能明显改善急性痛风性关节炎大鼠症状,减轻踝关节肿胀度,降低血清IL-1β、TNF-α及UA水平,对急性痛风性关节炎大鼠具有治疗作用。     

凝血酶法致大鼠血液高凝态模型的建立

目的探索静脉注射凝血酶造成大鼠高凝态模型,为高凝态的研究提供合适动物模型。方法 SD大鼠分为六组,分别于股静脉恒速注射生理盐水和2.5、5、10、20、40 U/kg凝血酶溶液,5 min内采血测活化部分凝血活酶时间(activated partial thromboplastin time,APTT)、凝血酶原时间(prothrombin time,PT)、纤维蛋白原(fibrinogen,FIB),并观察大鼠死亡情况以确定最佳凝血酶剂量。在此基础上,以最佳剂量凝血酶溶液股静脉注射给予大鼠,分别于0、10、30、60、120、180、300 s采血测APTT、PT、FIB以确定最佳采血时间。最后将大鼠分为生理盐水组与凝血酶组(最佳凝血酶剂量和采血时间),采集血样测定APTT、PT、FIB以及全血粘度。结果 10 U/kg凝血酶组大鼠血浆APTT、PT明显缩短,FIB明显升高,且大鼠死亡率低。注射凝血酶后60 s大鼠血浆APTT、PT缩短最多,FIB含量升至最高。同生理盐水组相比,凝血酶组大鼠血浆PT、APTT显著缩短,FIB、全血粘度显著增大,且差异有显著性(P0.05)。结论注射凝血酶溶液可复制大鼠血液高凝态模型,最佳凝血酶剂量为10 U/kg,凝血酶浓度为2U/m L,最佳采血测试时间为60 s。     

大蒜素对大鼠实验性大肠癌的作用及其机制研究

本研究旨在探讨大蒜素对实验性结肠癌大鼠的治疗作用及其作用机制。运用二甲肼30 mg/kg腹腔注射,每周注射1次,连续应用20周,成功建立实验性大鼠大肠癌模型,造模同时给予大蒜素注射液治疗。观察各组大鼠一般状态、脾脏指数、胸腺指数和组织病理学变化,酶联免疫吸附法(ELISA)检测血清中CD44V6和VEGF水平变化。Western blot法检测大肠组织中survivin、Cleaved Caspase-3和Cleaved Caspase-7的蛋白水平变化。实验结果显示第32周实验结束时,模型组大鼠平均体重与正常组相比显著降低(P0.05)。大蒜素高、低剂量组大鼠体重与模型组相比显著增加(P0.05)。大蒜素高、低剂量组大鼠平均胸腺指数、平均脾脏指数与模型组相比显著增高(P0.05)。而大蒜素高剂量组胸腺指数、脾指数显著高于大蒜素低剂量组(P0.05)。模型组肿瘤发生率、荷瘤数及肿瘤转移率均显著高于各给药组(P0.05)。模型组大鼠血清CD44V6和VEGF水平显著高于正常组(P0.05)。大蒜素高、低剂量组血清CD44V6和VEGF水平与模型组相比显著降低(P0.05)。而与低剂量组相比,大蒜素高剂量组二项指标水平增加更为显著(P0.05)。模型组大鼠大肠组织survivin、Caspase-3和Caspase-7表达水平显著高于正常组(P0.05)。但与模型组相比,大蒜素高、低剂量组survivin表达水平显著降低,而Cleaved Caspase-3和Cleaved Caspase-7表达水平显著升高(P0.05)。提示大蒜素对二甲肼诱导的实验性大鼠大肠癌具有良好的防治作用,可有效抑制大肠癌的发展及转移,其机制可能通过一方面增强大鼠体质,提高大鼠免疫功能,另一方面调节CD44V6和VEGF的水平来抑制肿瘤内血管的生成和浸润转移,同时抑制survivn基因的表达,上调Caspase-3和Caspase-7水平,从而促进癌细胞的凋亡。     

镉在大鼠前列腺组织内的蓄积

为比较重金属镉在去势与非去势雄性大鼠前列腺组织内蓄积量的变化,将60只健康雄性SD大鼠随机分为六组:对照组、去势对照组、低剂量镉组(2.5 mμmol·Kg^-1)、高剂量镉组(20 mμmol·Kg^-1)、去势低剂量镉(2.5 mμmol·Kg^-1)组、去势高剂量镉(20 mμmol·Kg^-1)组,18个月后取材进行实验.利用原子吸收分光光度法检测大鼠前列腺组织中重金属镉含量.与非去势对照组相比,各去势组大鼠前列腺组织内镉含量明显降低;注射镉后,大鼠前列腺组织镉含量明显高于对照组,但高、低剂量组之间无显著差别.实验表明长期镉暴露后,前列腺组织内镉含量明显增加;经去势手术的大鼠,长期镉暴露后前列腺组织内镉含量未升高,但机制并不明确.     

灌胃不同剂量钙对大肠肿瘤模型大鼠肿瘤及血氨基酸的影响的实验研究

目的:研究不同口服剂量钙对大肠肿瘤和血清中氨基酸代谢的影响。方法:50只雄性Wistar大鼠适应性喂养一周后随机分为5组每组10只,碳酸钙灌胃6个月。对照组(钙摄入量0.5 g/kg/d)、模型对照组(钙摄入量0.5 g/kg/d)、低剂量钙组(钙摄入量1.0 g/kg/d)、中剂量钙组(钙摄入量1.5 g/kg/d)和高剂量钙组(钙摄入量2.0 g/kg/d)。模型对照组及低中高钙剂量组在实验的第2周开始,颈部皮下连续注射二甲基肼20周进行大肠肿瘤造模。大鼠解剖后观察肿瘤数量和直径,亚甲蓝染色观察大肠内异常隐窝(ACF)数量,HE染色观察腺癌发生情况。液相色谱-质谱联用检测大鼠血液中相关氨基酸含量。结果:从第20周开始,与模型组相比,中剂量钙组的体重增加,有显著性差异(P0.05)。在肿瘤数、肿瘤发生率、肿瘤平均直径和肠重这些指标中,中剂量钙组和高剂量钙组均显著低于模型对照组(均P0.05)。病理组织学结果显示,正常对照组未见任何组织增生,模型对照组以腺瘤为主,出现小部分腺癌。而其余各组均以腺瘤为主,未出现腺癌。与模型对照组相比,中剂量钙组的谷氨酸、谷氨酰胺、鸟氨酸均显著升高。结论:钙灌胃对大鼠大肠肿瘤发生具有抑制作用,血清中谷氨酸、谷氨酰胺和鸟氨酸的代谢发生变化。     

羊骨酶解发酵液钙鳌合物对骨质疏松大鼠骨密度和骨代谢生化指标的影响

目的探讨羊骨酶解发酵液钙螯合物（SBEF-Ca）对雌激素缺乏造成的骨质疏松的防治作用。方法 40只3.5月龄的Sprague-Dawley大鼠,按体重随机分为正常组（假手术＋蒸馏水）、模型对照组（去卵巢＋蒸馏水）和高、中、低3个剂量组（去卵巢＋SBEF-Ca）。术后12 d,持续灌胃11周后,测定股骨密度、长度以及血清中反映骨代谢的主要生化指标。结果模型对照组大鼠股骨密度和长度显著减小,各剂量组均可扭转这种减小趋势并使股骨密度和长度维持在正常组水平,其中高剂量组的股骨密度显著大于中、低剂量组;各剂量组的股骨长度差异不显著;所有大鼠的血Ca、血P水平差异无显著性;模型对照组的血清碱性磷酸酶（ALP）、骨钙素（BGP）水平明显升高,而各剂量组的ALP、BGP水平与正常组差异不显著,且高剂量组ALP水平显著低于中、低剂量组,中、高剂量组的BGP水平（P〉0.05）显著低于低剂量组。结论低、中、高剂量的SBEF-Ca对绝经后骨质疏松症有预防作用。     

白藜芦醇对急性痛风性关节炎大鼠的影响(英文)

目的:观察白藜芦醇对急性痛风性关节炎大鼠的影响。方法:选取Wista大鼠36只,随机分为正常对照组、模型组、秋水仙碱组、白藜芦醇低剂量组、白藜芦醇中剂量组、白藜芦醇高剂量组,各组相应采用生理盐水、秋水仙碱、白藜芦醇低、中、高剂量灌胃7天(1次/日),模型组及各实验组于灌胃第四天把25g/mL(0.05mL)浓度的尿酸盐溶液注射到大鼠踝关节腔内,制备急性痛风性关节炎模型,正常对照组大鼠关节腔内注射生理盐水0.05 ml,72 h后留取踝关节关节液及关节滑膜,应用ELISA法观察关节液中IL-1β、CXCL10的变化。关节滑膜用10%福尔马林固定待做病理。结果:与模型组比较,白藜芦醇能显著降低关节液中IL-1β、CXCL10水平(P0.05),病理结果显示,白藜芦醇可减轻急性痛风性关节炎大鼠踝关节组织的水肿和炎性细胞浸润。结论:急性痛风性关节炎发病过程中IL-1β,CXCL10明显增高,白藜芦醇可有效抑制急性痛风性关节炎发作,且该作用呈一定的剂量依赖性。关键词:痛风性关节炎;白藜芦醇;白介素1-β     

补肾填精法对脑卒中失眠症模型大鼠血清褪黑素含量表达的影响

目的:探讨补肾填精法对脑卒中失眠症模型大鼠血清褪黑素(Melatonin,MT)含量表达的影响。方法:24只建模成功的脑卒中失眠症大鼠随机平分为3组-模型组、低剂量组、高剂量组。于造模第7 d起,三组分别给予生理盐水及补肾填精汤(低剂量组最终含生药浓度低剂量为0.7 g/mL、中剂量为1.4 g/mL)以1 mL/(100 g·d)灌胃,连续14 d,记录血清MT表达变化情况。结果:三组治疗第1 d、第7 d与第14 d的逃避潜伏期在组内与组间对比差异无统计学意义(P>0.05)。低剂量组、高剂量组治疗第7 d与第14 d的进入中央区次数多于模型组(P<0.05),也多于治疗第1 d(P<0.05),高剂量组也多于低剂量组(P<0.05)。低剂量组、高剂量组治疗第14 d的血清MT含量、去甲肾上腺素转运蛋白与褪黑素受体蛋白相对表达水平高于模型组(P<0.05),高剂量组高于低剂量组(P<0.05)。结论:补肾填精法在脑卒中失眠症模型大鼠中的应用能促进血清MT的释放,提高去甲肾上腺素转运蛋白与褪黑素受体蛋白的表达,从而促进缓解失眠症状。     

[PEAD:肝素:NGF]生物材料促进大鼠坐骨神经损伤恢复

目的:探讨新型材料poly(ethylene argininylaspartate diglyceride)(PEAD)结合肝素包裹神经生长因子组成的三元复合体比单纯运用NGF治疗大鼠坐骨神经损伤效果明显,为临床治疗外周神经损伤提供实验依据。方法:24只200g左右Wistar大鼠,分成生理盐水组,NGF组,NGF凝聚体三组,每组各8只,距梨状肌下缘远侧约1.5cm处运用静脉夹夹紧坐骨神经2min,采用无创细线(5/0)缝合肌肉和皮肤,并用碘伏进行消毒,NGF组每天沿坐骨切迹肌注80ngNGF,持续30天;NGF凝聚体组仅在造模时肌注复合体(内含2.4μg的NGF);生理盐水组给予等体积的生理盐水。术后每周运用脚步印迹法评价动物的行为学,并于30天后灌流、收集各组损伤侧坐骨神经,运用HE染色及投射电镜观察坐骨神经结构恢复情况,免疫荧光标记MBP,观察其蛋白的表达。结果:NGF组,NGF凝聚体组在行为学、病理结构及蛋白的表达远高于生理盐水组,并且NGF凝聚组的治疗效果优于NGF组。结论:新型凝聚体包载NGF具有明显的促进周围神经损伤后的修复与再生作用,能够在一定程度上提高单纯运用NGF治疗大鼠坐骨神经损伤的不足,达到更加理想和显著的促恢复效果。     

The Protective Effects of Achyranthes bidentata Polypeptides on Rat Sciatic Nerve Crush Injury Causes Modulation of Neurotrophic Factors

Pharmacological treatment is a therapeutic approach to improving nerve regeneration and functional recovery after peripheral nerve crush injury. The objective of the present study was to investigate the effects of the polypeptides isolated from Achyranthes bidentata Blume (abbreviated as ABPP) on rat sciatic crush injury and to test the possible involvement of neurotrophic factors. After surgical crush injury, rats received daily intraperitoneal injection of 0.2 ml saline containing 2 mg ABPP, 1 μg nerve growth factor (NGF) or no additive. The results from walking track analysis, electrophysiological assessment and histological evaluation indicated that the repair outcomes by ABPP treatment were close to those by NGF treatment, but better than those by treatment with saline alone. The quantitative real-time RT-PCR was used to monitor the mRNA expression of growth associated protein in the crush nerves and the mRNA expression of NGF, brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), tyrosine kinase (Trk)A and TrkB in the dorsal root ganglia (DRGs) at L4–L6. The mRNA expression of these genes in the crush nerve sample and DRGs sample was higher after treatment with ABPP or NGF than after treatment with saline alone. Our findings suggest that ABPP might protect peripheral nerve against crush injury through stimulating release of neurotrophic factors and the other cytokines.     

大鼠脊神经压迫后早期痛阈与神经损伤程度的联系

目的观察大鼠脊神经压迫后早期痛阈改变与神经损伤程度的联系。方法雄性SD大鼠32只,随机分为4组（每组n=8）：正常组,假手术组,70 g压迫组和180 g压迫组。正常组不作任何处理,作为对照,假手术组模型制作过程和其它两个模型组相同,但不做神经压迫,70 g压迫组和180 g压迫组分别用70 g或180 g血管夹压迫大鼠右侧颈7脊神经15 min,制作神经急性压迫损伤模型。于术前2 d和术后1、35、、7 d测右侧前足机械性痛阈和热痛阈,术后7 d处死大鼠取右侧颈7脊神经,HE染色病理分级和扫描电镜观察,比较各组痛阈改变以及神经损伤程度的变化。结果与术前相比较,正常组和假手术组术后机械性痛阈和热痛阈无明显变化（P〉0.05）;与术前以及正常组和假手术组相比,70 g压迫组术后1～7 d机械性痛阈和热痛阈明显降低（P〈0.05）,180 g压迫组术后1 d痛阈无明显变化（P〉0.05）,术后3～7 d明显升高（P〈0.05）。正常组和假手术组均未出现神经损伤病理变化,而70 g压迫组和180g压迫组出现不同程度的神经损伤,损伤病理分级180 g压迫组高于70 g压迫组（P〈0.05）。结论70 g或180 g血管夹压迫大鼠颈7脊神经后大鼠痛阈降低或升高,其机制可能与不同的压力导致不同的神经损伤程度有关。     

肌肉注射人神经生长因子基因逆轴突传递修复神经损伤的研究进展

神经生长因子(NGF)促进中枢及外周神经系统神经元细胞存活、分化、轴突再生等重要作用已得到临床的广泛证实。目前临床上主要以局部或肌肉注射NGF蛋白的方式对神经系统的损伤进行治疗。但NGF半衰期短、局部应用副作用大、费用昂贵、难以透过血脑屏障等缺点而限制临床应用。长期以来,科研工作者致力于寻求一种理想的途径或方法以克服这一缺陷。随着基因工程技术的飞速发展,研究人员发现通过骨骼肌肌肉注射途径,以非病毒载体介导外源的NGF基因体内表达并逆轴突传递到神经损伤部位,有望解决这一难题。本文将就NGF及受体的基本结构和特性、逆轴突传递的机制、非病毒载体结合骨骼肌肌肉注射的基因治疗等方面进行总结和阐述。     

Therapeutic Effect of Vinorine on Sciatic Nerve Injured Rat

Vinorine is a monoterpenoid indole alkaloid, a type of natural alkaloids. Growing reports exhibited the numerous pharmacology activities of vinorine such as anti-inflammation, anti-bacterial and anti-tumor. In this study, the effect of vinorine injection (7.5, 15 and 30 mg/kg) on motor function, sensation and nerve regeneration in sciatic nerve crush injury rat was investigated. The results of behavioral analysis, electrophysiological analysis and muscle histological analysis suggested that vinorine promoted the motor function recovery after sciatic nerve injury. The results of mechanical withdrawal thresholds assay and hot plate test demonstrated that vinorine improved the sensation recovery after sciatic nerve injury. The results of Fluoro-gold retrograde labeling, transmission electron microscope assay, toluidine blue and HE staining showed that vinorine attenuated the nerve damage caused by sciatic nerve injury and promoted the nerve regeneration. Furthermore, nerve growth factor (NGF) and its downstream extracellular signal-regulated kinase (ERK) signaling pathway participated in the neuro-recovery effect of vinorine after crush. In conclusion, vinorine treatment accelerated the sciatic nerve regeneration, motor function recovery and sensation recovery after crush injury via regulation of NGF and ERK activity. These results suggested that vinorine is a promising agent for never injury therapy.     

Neural Stem Cells Enhance Nerve Regeneration after Sciatic Nerve Injury in Rats

With the development of tissue engineering and the shortage of autologous nerve grafts in nerve reconstruction, cell transplantation in a conduit is an alternative strategy to improve nerve regeneration. The present study evaluated the effects and mechanism of brain-derived neural stem cells (NSCs) on sciatic nerve injury in rats. At the transection of the sciatic nerve, a 10-mm gap between the nerve stumps was bridged with a silicon conduit filled with 5?×?10⁵ NSCs. In control experiments, the conduit was filled with nerve growth factor (NGF) or normal saline (NS). The functional and morphological properties of regenerated nerves were investigated, and expression of hepatocyte growth factor (HGF) and NGF was measured. One week later, there was no connection through the conduit. Four or eight weeks later, fibrous connections were evident between the proximal and distal segments. Motor function was revealed by measurement of the sciatic functional index (SFI) and sciatic nerve conduction velocity (NCV). Functional recovery in the NSC and NGF groups was significantly more advanced than that in the NS group. NSCs showed significant improvement in axon myelination of the regenerated nerves. Expression of NGF and HGF in the injured sciatic nerve was significantly lower in the NS group than in the NSCs and NGF groups. These results and other advantages of NSCs, such as ease of harvest and relative abundance, suggest that NSCs could be used clinically to enhance peripheral nerve repair.     

Positive Effects of bFGF Modified Rat Amniotic Epithelial Cells Transplantation on Transected Rat Optic Nerve

PurposeEffective therapy for visual loss caused by optic nerve injury or diseases has not been achieved even though the optic nerve has the regeneration potential after injury. This study was designed to modify amniotic epithelial cells (AECs) with basic fibroblast growth factor (bFGF) gene, preliminarily investigating its effect on transected optic nerve.MethodsA human bFGF gene segment was delivered into rat AECs (AECs/hbFGF) by lentiviral vector, and the gene expression was examined by RT-PCR and ELISA. The AECs/hbFGF and untransfected rat AECs were transplanted into the transected site of the rat optic nerve. At 28 days post transplantation, the survival and migration of the transplanted cells was observed by tracking labeled cells; meanwhile retinal ganglion cells (RGCs) were observed and counted by employing biotin dextran amine (BDA) and Nissl staining. Furthermore, the expression of growth associated protein 43 (GAP-43) within the injury site was examined with immunohistochemical staining.ResultsThe AECs/hbFGF was proven to express bFGF gene and secrete bFGF peptide. Both AECs/hbFGF and AECs could survive and migrate after transplantation. RGCs counting implicated that RGCs numbers of the cell transplantation groups were significantly higher than that of the control group, and the AECs/hbFGF group was significantly higher than that of the AECs group. Moreover GAP-43 integral optical density value in the control group was significantly lower than that of the cell transplantation groups, and the value in the AECs/hbFGF group was significantly higher than that of the AECs group.ConclusionsAECs modified with bFGF could reduce RGCs loss and promote expression of GAP-43 in the rat optic nerve transected model, facilitating the process of neural restoration following injury.     

Nerve Growth Factor of Red Nucleus Involvement in Pain Induced by Spared Nerve Injury of the Rat Sciatic Nerve

Nerve growth factor (NGF), a member of the neurotrophin family, is essential for the development and maintenance of sensory neurons and for the formation of central pain circuitry. The current study was designed to evaluate the expression of NGF in the brain of rats with spared nerve injury (SNI), using immunohistochemical technique. The results showed that the level of NGF in the Red nucleus (RN) of SNI rats was apparently higher than that of sham-operated rats. To further study the effect of NGF in the development of neuropathic pain, different doses of anti-NGF antibody (20, 2.0 and 0.2 μg/ml) were microinjected into the RN contralateral to the nerve injury side of SNI rats. The data suggested that the higher doses of anti-NGF antibody (20 and 2.0 μg/ml) significantly attenuated the mechanical allodynia of neuropathic rats, while the 0.2 μg/ml antibody showed no analgesic effect. These results suggest that the NGF of RN is involved in the development of neuropathic allodynia in SNI rats.     

Recovery of Cholinergic Phenotype in the Injured Rat Neostriatum: Roles for Endogenous and Exogenous Nerve Growth Factor

Polyclonal antibodies against recombinant human nerve growth factor (rhNGF) potently inhibited PC12 neurite outgrowth, blocked high-affinity 125I-rhNGF binding but not its receptor, and cross-reacted with rat, mouse, and human nerve growth factor (NGF) but not with brain-derived neurotrophic factor, neurotrophin-3, ciliary neurotrophic factor, insulin-like growth factor, epidermal growth factor, or activin A. Immunocytochemistry revealed many NGF-positive neurons in the rat neostriatum. The NGF-positive neurons disappeared by 3 days after mechanical injury to the neostriatum and were replaced by intensely NGF- and glial fibrillary acidic protein-positive astrocytes. Enzyme-linked immunosorbent assay measurements revealed that the NGF content of the injured striatum was elevated by eightfold 3 days postinjury and by twofold 2 weeks later. The high-affinity choline uptake (HACU) into cholinergic nerve terminals was decreased by 23% at 2 and 4 weeks postinjury, yet choline acetyltransferase (ChAT) activity in these neurons was unchanged at 2 weeks and decreased by 14% at 4 weeks. Daily infusion of 1 microgram of rhNGF into the injury area did not alter the loss of HACU. However, this treatment elevated ChAT activity by 23-29% above intact neostriatal levels and by 53-65% relative to HACU at both survival times. Thus, lesion-induced increases in NGF levels within astrocytes are associated with maintenance of striatal ChAT activity at normal levels following cholinergic injury, even with decreases in HACU. Pharmacologic doses of rhNGF can further augment ChAT activity in damaged cholinergic neurons, showing the usefulness of exogenous NGF even when endogenous NGF is elevated in response to injury.     

Synergistic Effects of Bone Mesenchymal Stem Cells and Chondroitinase ABC on Nerve Regeneration After Acellular Nerve Allograft in Rats

This study aimed to evaluate whether combination therapy of bone marrow stromal cells (BMSCs) transplantation and chondroitinase ABC (ChABC) treatment further enhances axonal regeneration and functional recovery after acellular nerve allograft repair of the sciatic nerve gap in rats. Eight Sprague–Dawley rats were used as nerve donors, and 32 Wistar rats were randomly divided into four groups: Group I: acellular rat sciatic nerve (ARSN) group; Group II: ChABC treatment; Group III: BMSCs transplantation; and Group IV: ChABC treatment and BMSCs transplantation. The results showed that compared with ARSN control group, BMSC transplantation promoted axonal regeneration, the secretion of neural trophic factors NGF, BDNF and axon angiogenesis in nerve graft. ChABC treatment degraded chondroitin sulfate proteoglycans in ARSN in vitro and in vivo and improved BMSCs survival in ARSN. The combination therapy caused much better beneficial effects evidenced by increasing sciatic function index, nerve conduction velocity, restoration rate of tibialis anterior wet muscle weight, and myelinated nerve number, but did not further boost the therapeutic effects on neurotrophic factor production, axon angiogenesis, and sensory functional recovery by BMSC transplantation. Taken together, for the first time, we demonstrate the synergistic effects of BMSC transplantation and BMSCs treatment on peripheral nerve regeneration, and our findings may help establish novel strategies for cell transplantation therapy for peripheral nerve injury.