靶肌肉注射甲钴胺对大鼠周围神经损伤再生的作用

目的　通过靶肌肉 (腓肠肌 )注射不同剂量的甲钴胺 ,观察其对大鼠坐骨神经损伤的再生作用 ,为临床肌肉局部注射甲钴胺促进周围神经再生探讨较佳的用药剂量和方法。　方法选用健康Wistar大鼠 30只 ,左侧坐骨神经横断后即刻行外膜缝合 ,制备大鼠坐骨神经损伤模型。随机分为A、B、C三组 ,每组 1 0只 ,A组注射甲钴胺 30 0 μg kg,B组注射甲钴胺 1 0 0 μg kg ,C组注射同体积的等渗盐水 1ml,术后靶肌肉注射 1次 d。术后第 4周、第 8周每组分别提取 5只大鼠 ,以左小腿三头肌湿重、光镜和电镜坐骨神经形态学观察并图像分析作为检测指标 ,分析不同剂量甲钴胺对大鼠坐骨神经损伤的影响。　结果　术后 4周左小腿三头肌湿重 ,A组为 (1 .36 7± 0 .0 1 2 )g ,B组为 (1 .1 6 4± 0 .0 1 1 )g,C组为 (0 .95 0± 0 .0 0 9)g ,差异有显著性意义 (P <0 .0 5 ) ;术后 8周 ,A组为 (2 .2 0 5± 0 .0 1 5 )g ,B组为 (1 .6 1 1± 0 .0 1 3)g ,C组为 (1 .2 30± 0 .0 1 4 )g ,差异有显著性意义 (P <0 .0 5 )。术后 4周和 8周行光镜和电镜观察 ,图像分析坐骨神经髓鞘的横截面积、壁厚度 ,A组优于B组 ,B组优于C组。　结论　靶肌肉注射甲钴胺可以促进周围神经的再生 ,高剂量的甲钴胺可以更好地发挥其对损伤神经的营养诱导作用 ,值     

神经生长因子及三磷酸腺苷联合应用修复大鼠周围神经损伤及对失神经肌肉的作用

目的 探讨腹腔联合应用鼠源性神经生长因子(NGF)及三磷酸腺苷(ATP)对周围神经损伤修复及失神经肌肉的作用. 方法 选取SD大白鼠96只,体重(250±15)g,雌雄不限,按随机数字表法分为等渗盐水组(NS组)、ATP组、NGF组、ATP+NGF组;选取大鼠坐骨神经,暴露后人为切断造成3 mm缺损损伤,通过硅胶管连接两神经断端,建立神经再生室.术后立即对各组分别予以腹腔内注射等渗盐水0.4 ml、ATP0.4 ml(1 mg/ml)、NGF0.4 ml(0.2 μg/ml)、NGF+ATP 0.4 ml(ATP 1 mg/ml、NGF 0.2 μg/ml),之后每3 d通过腹腔注射1次.在第2,4,6,8周各组分别取6只大门鼠行坐骨神经指数测定、大体观察、坐骨神经肌电图检查,处死大鼠后切取标本观察再生室区段神经再生状况、腓肠肌湿重,HE染色观察腓肠肌纤维直径及截面积. 结果 术后第2,4,6,8周各组分别取6只大白鼠测定实验数据,NGF+ATP组坐骨神经指数、神经传导速度、腓肠肌湿重优于NS组及单纯ATP或NGF组(P＜0.05).大体观察可见NGF+ATP组患侧足趾红肿程度较其余三组轻,局部溃烂亦相对少,未见足趾脱落者.术后2周,各组神经再生室内可见纤细神经纤维生长,但未将神经断端连接一起.术后4,6,8周,神经再生室内可见神经纤维通过,NGF+ATP组相对其余三组神经纤维生长生长情况较好,且局部炎性反应及纤维粘连轻.NGF组神经纤维生长情况好于ATP组,但二者均优于NS组.第2,4,6,8周神经电生理及腓肠肌湿重NGF组优于ATP组,第4,6,8周坐骨神经指数、肌纤维直径及截面积NGF组优于ATP组(P＜0.05),且各时相点NGF组及ATP组的观测指标均明显优于NS组(P＜0.05). 结论 (1)联合应用NGF+ATP对神经损伤修复及失神经肌肉作用明显,优于单纯应用NGF及ATP;(2)单纯应用ATP对神经损伤修复及失神经肌肉有一定作用,但效果劣于NGF及NGF+ATP.     

碱性成纤维细胞生长因子促进周围神经功能恢复的研究

目的 探讨碱性成纤维细胞生长因子对受损伤的周围神经功能恢复的影响。方法 以钳夹损伤左侧坐骨神经大鼠为模型，治疗组肌肉注射ｂＦＧＦ，对照组注射等渗盐水，隔日１次。术后２，３，４周分别对治疗组和对照组损伤侧及健侧坐骨神经，腓肠肌进行功能检测，以健侧为１００％，求出损伤侧各指标的恢复率。     

大鼠坐骨神经损伤后显微及超微结构的三维重建

利用大鼠坐骨神经损伤后再生神经组织连续切片重建其显微及超微结构的三维图像。用硅胶管桥接90只大鼠左侧坐骨神经长6mm缺损，术后7、15、30、60、90天取坐骨神经行光、电镜检查，经显微摄像和扫描仪采集输入微机进行图像配准和分割，采用直接体素绘制模型实现三维重建和显示。结果显示，利用光镜和电镜图像完成了坐骨神经损伤后变性、再生过程中神经纤维及附属结构的形态变化的三维图像重建。重建结果能直观、形象地揭示大鼠再生坐骨神经纤维与正常神经纤维结构形态的差异，可作为周围神经损伤研究的一种新手段。     

动脉套管联合聚乙二醇在大鼠周围神经损伤中的应用

目的 观察应用动脉套管联合小间隙缝合聚乙二醇(PEG)冲洗法修复大鼠周围神经损伤的效果.方法 建立大鼠坐骨神经离断后神经重建模型,144只大鼠平均分为4组,每组36只,均选用雄性SD大鼠,体重200～250g.其中坐骨神经断端采用小间隙缝合盐水冲洗修复为A组,小间隙缝合PEG冲洗修复为B组,动脉套管联合小间隙缝合盐水冲洗修复为C组,动脉套管联合小间隙缝合PEG冲洗修复为D组.评估神经功能恢复时,采用行为学坐骨神经功能指数(SFI)及电生理神经冲动传导与复合肌肉动作电位(CMAP)对4组数据进行分析.评估神经再生情况时,应用苏木精-伊红染色法(HE染色)和透射电子显微镜(TEM)对4组数据进行采集.所有数据在各组术后1、4和8周进行检测.结果 D组行为学SFI值显著优于其他组;电生理学示各组在术后所有时间点均产生混合肌肉运动电位CMAP,但D组运动电位峰值高于其他组;组织病理学HE染色示D组下肢肌肉萎缩程度小于其他组且在对坐骨神经进行染色时发现D组再生神经纤维排列有序,具有正常直径和施万细胞,成纤维细胞较少;TEM示D组再生神经纤维数量最多、轴突最厚、髓鞘外观正常.结论 在对大鼠坐骨神经损伤模型中应用动脉套管联合小间隙缝合PEG冲洗修复效果显著优于其他方法,通过后期的临床转化,有望为临床上周围神经损伤的修复提供一种新的思路.     

重组腺病毒介导肝细胞生长因子基因治疗大鼠肝纤维化的实验研究

目的:评价携带人肝细胞生长因子(HGF)基因的重组腺病毒对大鼠肝纤维化模型的治疗作用。方法:DMN法建立W istar大鼠肝纤维化模型后,将8.6×109pfu的Ad-HGF经尾静脉注射入大鼠体内,对照组注射同剂量的Ad-GFP。第40天心脏采血,检测血清GPT、GOT、TP和TB il,并取肝组织做HE染色,观察两组大鼠肝脏的病理形态。结果:治疗组部分动物肝功能有所改善,对照组不明显。与对照组相比,治疗组大鼠的肝组织损伤程度较轻,汇管区看不到明显的纤维增生,并可见双核肝细胞和分裂相细胞。结论:腺病毒介导的HGF基因可能有部分治疗大鼠肝纤维化的作用。     

肝细胞生长因子在心血管疾病中的作用机制及应用研究进展

肝细胞生长因子(hepatocyte growth factor, HGF)是由69kD的α链及34kD的β链组成的异二聚体蛋白[1-2],因最初发现其活性与肝细胞再生有关,故名肝细胞生长因子,后研究发现HGF是一种可以作用于多种组织与细胞的生长因子,具有促进细胞增殖、细胞运动、形态发生、血管新生、抑制凋亡等作用.HGF的主要来源有成纤维细胞、平滑肌细胞、肥大细胞、巨噬细胞和白细胞等[3].IL-1、PDGF、bFGF和G-CSF等可诱导HGF的表达,TGF和糖皮质激素等则抑制其表达[3].     

联合应用神经生长因子和睫状神经营养因子治疗大鼠坐骨神经损伤

目的 探讨联合应用神经生长因子(NGF)和睫状神经营养因子(CNTF)对大鼠坐骨神经损伤后再生及功能恢复的影响。方法 切除120只大鼠左侧6mm长坐骨神经，随机分为4组，每组30只，分别行NGF CNTF、CNTF、NGF、等渗盐水(NS)靶肌肉注射。伤后行坐骨神经功能指数(SFI)、形态学和S—100α、神经中丝200(NF200)免疫组化检查。结果 联合应用NGF和CNTF组SFI有髓神经纤维直径、数目和S—100α、NF200阳性轴突计数均明显优于单独用药组。结论 联合应用NGF和CNTF可明显促进大鼠坐骨神经损伤后的再生与功能恢复。     

坐骨神经挤压模型大鼠脊髓背角c-fos表达的研究

目的 建立大鼠坐骨神经挤压模型,观察其神经恢复过程中行为学、机械痛阈值、热缩足反射潜伏期(TWL)以及脊髓背角c-fos表达(对c-fos表达产物:Fos蛋白阳性的神经元进行计数)的改变,探讨外周神经损伤再生与痛觉过敏的关系.方法 雄性Wistar大鼠40只,体重180～200g,随机分为坐骨神经血管钳挤压损伤组(A组)和假手术组(B组),每组20只.前者分离出右侧坐骨神经后于神经中段用止血钳挤压神经直到成为透明膜状,持续30s;后者将坐骨神经于空气中暴露5min后,还复至原位.术后第18、21、24、27、30、33、36、39天重复检测大鼠行为学、机械痛阈值、TWL值.术后21、27、33、39d处死大鼠,免疫组化检测Fos蛋白表达.结果 A组术后各时间点的累积疼痛评分均高于B组(P＜0.05),但A组内各时间点间无差异.A组术后21d后机械痛阈值和TWL均低于B组(P＜0.05).A组术侧L4-5.脊髓背角浅层Fos蛋白阳性反应神经元计数大于B组(P＜0.05).A组39d的机械痛阈和TWL值均大于21d(P＜0.05),但Fos蛋白阳性反应性神经元计数无显著差异.结论 坐骨神经挤压模型中外周神经损伤再生过程诱发脊髓背角浅层c-fos表达,同时出现再生神经支配区域的机械痛觉过敏和疼痛相关行为.     

复合全氟三丁胺携氧材料的嗅鞘细胞移植促进大鼠周围神经损伤修复的研究

目的探究复合全氟三丁胺（perfluorotributylamine,PFTBA）的嗅鞘细胞（olfactory ensheathing cells,OECs）移植能否更进一步促进外周神经损伤后神经再生以及功能恢复。方法从大鼠嗅球分离并纯化OECs并鉴定。制作大鼠坐骨神经缺损模型。将60只坐骨神经缺损大鼠随机分为3组：自体神经移植组,OECs神经导管组,PFTBA-OECs神经导管组。在体内环境下分别测试术后3、7、14、28 d PFTBA对OECs存活率的影响。术后4周,对再生神经进行免疫荧光染色以及超薄切片,确定PFTBA-OECs对再生神经的作用。并在术后2、4、8周分别对各实验组大鼠坐骨神经功能指数值进行测量,以确定神经功能恢复情况。结果分离纯化的原代OECs纯度为(95.8±2.1)%;体内条件下,PFTBA能够大幅提高OECs的存活率（P<0.05）;与OECs神经导管组比较,PFTBA-OECs神经导管组极大地促进了神经轴突的再生（P<0.05）;再生神经透射电镜显示,PFTBA-OECs神经导管组再生神经轴突数量明显多于OECs神经导管组,髓鞘厚度明显优于OECs神经导管组（P<0.05）;术后坐骨神经功能指数值显示,PFTBA-OECs神经导管组大鼠功能恢复情况明显优于OECs神经导管组（P<0.05）。结论PFTBA能够明显提升OECs的促神经修复能力,明显提升神经损伤后的轴突再生能力以及髓鞘化程度,为解决细胞移植后面临的缺氧微环境提供了解决方案。     

碱性成纤维细胞生长因子(bFGF)对周围神经再生作用的实验研究

目的:探讨外源性bFGF对周围神经再生的作用。方法:6O只SD大鼠随机分治疗组、对照组各30只,钳夹损伤右侧坐骨神经后每日分别给予bFGF和生理盐水,术后行神经电生理和组织学检查。结果:计量分析治疗组运动神经传导速度、神经纤维密度、轴突直径和髓鞘厚度明显优于对照组。结论:bFGF能促进周围神经再生。     

睫状神经营养因子促进大鼠周围神经再生的实验研究

目的 探讨局部应用睫状神经营养因子（ＣＮＴＦ）对大鼠周围神经损伤后再生的影响。方法 ３４只大鼠分为４组,１￣３组各１０只,切除两侧坐骨神经各１０ｍｍ并用硅胶管套接,左侧套管内注入ＣＮＴＦ５０μｇ,右侧注入等渗盐水对照。术后１,３,４个月分别进行电生理和组织学检查。另４只鼠于术后３个月行辣根过氧化物酶（ＨＲＰ）逆行示踪检查。结果 ＣＮＴＦ治疗组术后１,３,４个月时的电生理和组织学检查结果均明显优于对     

Involvement of Cdc2 in axonal regeneration enhanced by exercise training in rats

PURPOSE: Physical activity can improve sensorimotor recovery after peripheral nerve injury. We examined the effects of treadmill training (TMT) on axonal regeneration in the injured sciatic nerve of the rat and further investigated cellular and molecular events that underlie enhanced axonal regrowth by training. METHODS: After crush injury of the sciatic nerves, rats were randomly assigned into either TMT or sedentary groups. Three to 14 d after injury, changes in protein levels in the regenerating nerve were analyzed by Western blotting and immunofluorescence staining. Axonal regeneration was assessed by anterograde and retrograde tracing techniques. The animals' functional recovery was determined by the sciatic functional index. RESULTS: We identified enhanced axonal regrowth in the distal stump of the sciatic nerve 7-14 d after injury in the rats with TMT. Cell division cycle 2 (Cdc2) mRNA and protein levels were highly increased in the injured sciatic nerves 3 and 7 d after injury, and decreased to basal levels 14 d later. Daily TMT accelerated distal shift of Cdc2 mRNA and protein induced in the regenerating nerves, and Cdc2 kinase activity was similarly increased in the distal stump by TMT. Cdc2 protein induced by TMT was mainly colocalized with Schwann cell marker S100beta protein, and correlated with axial distribution pattern of bromodeoxyuridine-labeled proliferating cell population in the regenerating nerve. We further demonstrate that axonal regeneration and motor function recovery after injury, both of which were promoted by TMT, were greatly suppressed by in vivo administration of Cdc2 inhibitor roscovitine. CONCLUSION: The present data suggest that Cdc2 kinase activated in the regenerating sciatic nerve may play an important role in TMT-mediated enhancement of axonal regeneration.     

缝线黏附重组pcDNA3-GDNF-GFP质粒在周围神经损伤中的实验研究

目的 研究周围神经损伤后新的修复方法,了解携带含胶质细胞源性神经营养因子（GDNF）基因重组质粒的缝合线对损伤的周围神经修复的影响.方法 使用prolene线浸泡于0.1%的左旋多聚赖氨酸和质粒等体积液并冻干,反复3次后使缝线黏附上质粒DNA.50只雌性Wistar大鼠制成右侧坐骨神经缺损动物模型,随机分为实验组和对照组,每组各25只.实验组使用黏附pcDNA3-GDNF-GFP的缝线,对照组使用黏附pcDNA3的缝线进行坐骨神经的缝合修复.术后1 w、8 w使用激光共聚焦显微镜检测GDNF在神经修复处的表达,于坐骨神经吻合处取材行免疫组化检测;手术后12 w行右侧坐骨神经电生理检查.结果 携带pcDNA3-GDNF-GFP重组质粒的缝线修复周围神经时,该质粒可以转染周围神经并得到表达.实验组神经的传导速度、动作电位的峰值及潜伏期分别是（19.82±1.90）m/s、（3.47±0.32）mV、（0.36±0.03）ms,明显优于对照组.结论 使用携pcDNA3-GDNF-GFP重组质粒的缝线修复损伤的周围神经,可以促进损伤神经的再生.     

Magnetic resonance imaging evaluation of acute crush injury of rabbit sciatic nerve: correlation with histology

OBJECTIVE: To investigate the relation between the quantitative assessment of magnetic resonance imaging (MRI) features and the correlation with histology and functional recovery by using the rabbit sciatic nerve crush model. METHODS: In New Zealand, 32 rabbits were randomly divided into 2 groups (group A and B); all rabbits underwent crushing injury of their left sciatic nerve. In group A (n = 16), the sciatic nerves were crushed by using microvessel clamps with a strength of 3.61 kg. In group B (n = 16), the sciatic nerves were crushed with a strength of 10.50 kg. Right sciatic nerves were served as controls. Serial MRI of both hind limbs in each rabbit was performed before and at the time point of 1, 2, 4, and 8 weeks after crushed injury. The MRI protocol included T1-weighted spin-echo (T1WI), 3 dimension turbo spin-echo T2-weighted (3DT2WI), T2-weighted turbo spin-echo images with spectral presaturation with inversion recovery (T2WI/SPIR), balanced fast-field echo (B-FFE) and short-time inversion recovery (STIR) sequences. The coronal image of the sciatic nerve was obtained. The nerve and muscle signal ratio (SIR) on each sequence was measured. The function recovery was observed and pathological examination was performed at each time point. RESULTS: A signal intensity increase of the distal segment of crushed sciatic nerves was found on 3DT2WI, T2WI/SPIR, B-FFE, and STIR, but not on T1WI images. Of 32 crushed nerves, 30 nerves showed high signal intensity. The correct diagnostic rate was 93.75% with false negative-positive of 6.25%. The SIR of the crushed sciatic nerve at distal portion was higher than those of the control nerves; there was a statistically significant difference (P < 0.001). The SIR of the distal portion of crushed nerves was higher than that of the proximal nerve portion; there was a statistically significant difference (P < 0.001). Whereas, the SIR at proximal nerve portions of crushed nerve was similar to control nerves (P > 0.05). The SIR between group A and group B was not found statistically significantly different (P > 0.05). The SIR of crushed nerves at distal portion increased at one week after the crush injury, subsequently further increased, and reached a maximum at 2 weeks. The pathological examination revealed myelin swelling and axonal fragmentation of crushed nerve. Abduction function of injured hind limb was deficit. From 4 to 8 weeks following the crush, the SIR decreased, correspondingly, nerve regeneration was revealed on pathology including extensive Schwann cells proliferation and the immature myelin formation. The abduction function gradually recovered. There was no abnormal finding on MRI for control and sham-operated nerves. CONCLUSION: The SIR of injured nerve at distal portion increased on MRI. The evolution of SIR after injury was correlated with the degeneration and regeneration of nerve and the function recovery of lower extremities. Assessment of peripheral nerve injury by using SIR could reveal acute nerve injury, as well as aid in monitoring the recovery process. The pathophysiological basis for the SIR is predominantly the results of axon breakdown and myelin regeneration     

外源性神经生长因子对神经在冻融肌移植体中再生的影响

采用在冻融骨骼肌桥接物中加入外源性神经生长因子的方法，修复兔坐骨神经缺损２．０ｍｍ。应用电生理学、组织学、免疫组织化学、图像分析、透射电镜等测试方法，发现早期神经轴突长入桥接体的速度及成熟程度明显优于单纯自体冻融骨骼肌桥。为探索非神经组织桥接神经再生，及自体变性骨骼肌桥接物的临床应用，提供了基础实验依据。     

高频超声对兔坐骨神经急性挤压伤的观察

目的：运用高频超声观察兔坐骨神经急性挤压伤，探讨其临床诊断价值。方法：按观察时间的不同将16只健康家兔随机分为4组。建立兔坐骨神经急性挤压伤模型，损伤后第1、2、4、8周，分别应用高频超声在同一水平上观察双侧坐骨神经的变化。结果：坐骨神经挤压伤后，其声像图和内径均有变化，且与神经损伤后退变、再生及肢体功能的动态变化相一致。结论：高频超声可直观地反映神经退变和再生的过程，提供了诊断外周神经损伤的新方法，对临床判断和预后提供客观依据。