乳鼠坐骨神经条件培养基促进周围神经再生的实验研究

通过对乳鼠坐骨神经条件培养基的研究，了解其对周围神经再生的作用。方法：用静脉桥接１０ｍｍ缺损的鼠坐骨神经，静脉腔内注入用Ａｓｋａｎａｓ植块法得来的乳鼠坐骨神经条件培养基，对照组则注入单纯的培养液。术后８周，１２周进行电生理，光镜及图象分析的研究。     

银杏叶提取物促进大鼠坐骨神经再生的实验研究

研究显示 ,银杏叶提取物 (ｅｘｔｒａｃｔｏｆｌｅａｖｅＧｉｎｋｇｏｂｉｌｏｂａ,ＥＧｂ)具有良好的神经保护作用[1 4 ] 。我们进行了ＥＧｂ对神经再生影响的研究。1.材料与方法 :雄性ＳＤ大白鼠 48只 ,随机分成损伤实验组 (ｎ =2 4)和对照组 (ｎ =2 4) ,ＥＧｂ由美国ＰＨＡＲＭＡＮＥＸＩＮＣ公司提供。制作坐骨神经挤压伤模型 ,术后对照组给予生理盐水 2ｍｌ灌胃 ,实验组给予ＥＧｂ 2 0ｍｇ (溶于 2ｍｌ生理盐水中 )灌胃。分别于术后第 2、4、6周作以下观察 :坐骨神经功能指数(ＳＦＩ) ;电生理检测 :用ＥｓａｏｔｅＰｈａｓｉｓ肌电仪测…     

外源性表皮生长因子促端侧吻合神经再生的研究

目的　评价外源性表皮生长因子对端侧吻合后神经远段再生的影响。　方法　选用雄性Wistar大白鼠 32只 ,随机分成 2组 ,每组 16只。EGF组 :切断右侧腓总神经 ,在邻近的胫神经干外膜上开一直径为 1mm小窗 ,将腓神经远端吻合到胫神经干侧方开窗处。右小腿外侧肌内每天注入生理盐水稀释的EGF注射液 ( 2 μg·μl-1) 0 1ml ,共用 2周 ;对照组 :吻合方法同EGF组 ,术后注射等量的生理盐水。各组分别于术后 4周、8周进行大体、组织学、形态定量学和电生理检测。　结果　端侧吻合加EGF组在各时间点 ,吻合口远段再生神经的数目、有髓神经纤维截面积、运动神经传导速度均明显优于对照组 (P <0 0 5 )。超微结构观察 :EGF组再生神经的有髓纤维数、髓鞘厚度和髓鞘的成熟度明显优于对照组。　结论　外源性表皮生长因子可以促进端侧吻合后神经远段再生和功能的恢复。     

促红细胞生成素促进大鼠坐骨神经再生作用的实验研究

目的 探讨促红细胞生成素(Erythoropoietin,EPO)对大鼠坐骨神经损伤修复后神经再生的影响,为周围神经损伤的临床治疗提供实验依据.方法 雄性SD大鼠40只,随机分为两组,即EPO组和神经生长因子(NGF)组,用硅胶管桥接10 mm的坐骨神经缺损,EPO组和NGF组分别注射EPO和NGF.术后4周和8周时每组分别提取10只大鼠,以坐骨神经功能指数(SFI)、运动神经传导速度(MNCV)、形态学观察和蛋白基因产物9.5(PGP 9.5)免疫组织化学染色,评估EPO对大鼠坐骨神经再生的影响.结果 术后4周SFI,EPO组为[(-78.85±3.87),x-±s,下同],NGF组为(-79.98±4.58),差异无统计学意义(P>0.05);术后8周SFI,EPO组为(-60.26±2.91),NGF组为(-64.65±4.11),差异有统计学意义(P<0.05).术后4周和8周时,EPO组MNCV、有髓神经纤维数目以及PGP9.5免疫阳性神经纤维的平均光密度和积分光密度均优于NGF组,差异有统计学意义(P<0.05).结论 EPO 能促进大鼠坐骨神经损伤后的修复与再生.     

不同强度直流电刺激器对鼠坐骨神经再生的影响

不同强度直流电刺激器对鼠坐骨神经再生的影响孟庆刚，于钟毓，岳琦，杨桂兰，孙立国，邢燕，李文庆尽管目前已证实直流电刺激可有效地促进鼠周围神经损伤的再生，但有关最佳刺激电流强度的确定仍是实验中尚未解决的难题。本实验通过应用不同强度全置入式弱直流电刺激器，...     

生长因子促进转基因小鼠坐骨神经体外预变性的实验研究

目的 探索一种简便高效的小鼠坐骨神经体外预变性的方法,以便在短期内获取大量高活性的雪旺细胞.方法　选用绿色荧光蛋白(GFP)转基因的小鼠24只,随机分为A、B、C和D四组.A、B、C组切取双侧坐骨神经,分别置于DMEM、DMEM+10％FBS及含生长因子的雪旺细胞培养液中体外预变性1周,D组切断一侧坐骨神经体内预变性1周.1周后取材,行免疫组织学和细胞学方面的检测,比较各组获取的雪旺细胞量和纯度.结果　C组获得的雪旺细胞量及纯度较高,与D组的雪旺细胞相似.结论　生长因子能促进小鼠坐骨神经的体外预变性,具有与体内预变性相似的效果,能在短期能获得大量雪旺细胞.     

甘油保存鼠坐骨神经异体移植诱导神经再生的研究

为制备既无明显抗原性.又具备完整神经内膜管结构,且便于长期保存的神经移植物.我们用甘油保存鼠坐骨神经异体移植.诱导神经再生.现将实验结果报道如下.     

人羊膜上皮细胞对大鼠坐骨神经再生影响的实验研究

目的 通过实验研究了解人羊膜上皮细胞(human amniotic epithelial cells,HAECs)对大鼠坐骨神经再生的促进作用.方法 取60只SD大鼠随机分为两组,羊膜细胞组和对照组,各组再分为2周和4周组,每组15只.制作大鼠周围神经损伤再生室模型,HAECs组局部应用培养的HAECs,对照组局部使用等量的生理盐水.术后分别于2周、4周检测神经传导功能和HE染色观察神经纤维形态学变化.结果 术后2周两组的神经电生理及组织学检测比较差异无统计学意义;4周HAECs组的大鼠坐骨神经传导功能明显恢复,HE染色显示术后坐骨神经手术部位出现大量炎性肉芽组织,呈现纤维性修复,吻合口以远HAECs组神经纤维形态结构较对照组完整,神经纤维与髓鞘直径均较对照组大.结论 HAECs移植可加速大鼠坐骨神经损伤后神经传导功能恢复,有助于有髓神经纤维损伤后的轴突与髓鞘的再生修复.

Abstract：

Objective To study the effects of human amniotc epithelial cells (HAECs) on regeneration of rat sciatic nerve. Methods Sixty SD rats were randomly divided into two groups, the HAECs group and control group. Each group was further divided into two week group and four week group, with 15 rats each. The sciatic nerve was cut and the regeneration chamber was created. The HAECs were implanted into the regeneration chamber in the HAECs group and normal saline was injected into the regeneration chamber in the control group.Electro-physiological and the histological study was done at two weeks and four weeks after the surgery. The results of nerve conduction study and HE staining of regenerating nerve fibers were analyzed statistically.Results There were no statistically significant differences between the two groups at two weeks post-operatively.At four weeks, the experimental group showed significantly higher nerve conduction velocity and amplitude of evoked potentials comparing to those of the control group. Nerve fibers and inflammatory granulation tissue was seen near the lesion site by HE staining in the control group, whereas the HAECs group showed more integrated nerve fihers and mature myelination distal to the lesion site. Conclusion The application of human amniotic epithelial cells may enhance nerve regeneration in rats.     

表皮生长因子促进皮肤创面愈合的研究

为了观察外源性表皮生长因子对创面愈合的影响，在３０例大鼠皮肤创面愈合模型中，局部外用表皮生长因子（ＥＧＦ），以软膏基质作为对照，分别于１，２周测量创面面积，测定创面组织中的ＤＮＡ、蛋白质及羟脯氨酸含量，记录创面完全愈合时间。结果表明，实验组与对照组创面完全愈合的时间分别为（１４．６±１．２）天和（１８．５±２．０６）天（Ｐ＜０．０１），ＥＧＦ能显著增加组织中的ＤＮＡ、蛋白质及羟脯氨酸含量（Ｐ＜０．０１）。结果提示，ＥＧＦ能显著加速创面的修复，缩短愈合时间。     

Vascular endothelial growth factor promotes peripheral nerve regeneration after sciatic nerve transection in rat

Objective：To evaluate the local effect of vascular endothelial growth factor （VEGF） on transected sciatic nerve regeneration.Methods：Sixty male white Wistar rats were divided into four experimental groups randomly （n=15）.In transected group the left sciatic nerve was transected and the stump was fixed to adjacent muscle.In treatment group the defect was bridged using a silicone graft filled with 10μL VEGF.In silicone group the graft was filled with phosphate-buffered saline.In sham-operated group the sciatic nerve was exposed and manipulated.Each group was subdivided into three subgroups with five animals in each and nerve fibers were studied 4,8 and 12 weeks after operation.Results：Behavioral test,functional study of sciatic nerve,gastrocnemius muscle mass and morphometric indices confirmed a faster recovery of regenerated axons in VEGF group than in silicone group （P〈0.05）.In immunohistochemical assessment,reactions to S-100 in VEGF group were more positive than that in silicone group.Conclusion：Local administration of VEGF will improve functional recovery and morphometric indices of sciatic nerve.     

外源性神经生长因子对内源性神经生长因子的影响

目的：探讨外源性神经生长因子对内源性神经生长因子的影响，方法：采用大鼠坐骨神经硅管内再生室模型，靶器官内注射神经生长因子（实验组）或生理盐水（对照组）２周，注射后１天，１周天及２周，采用新生大鼠背根神经节细胞培养，测定硅管内液神经营养活性，用化学发光免疫检测法（ｃｈｅｍｉｌｕｍｉｎｅｓｃｅｎｃｅｉｍｍｕｎｏａｓｓａｙ，ＣＬＩＡ）测定神经生长因子含量，结果：实验组与对照组细胞存活数及神经生长因子含量     

电针对大鼠坐骨神经再生影响的功能评价

目的从功能恢复评价电针对神经再生的促进作用。方法建立大鼠坐骨神经损伤模型，将９０只ＳＤ大鼠随机分为损伤对照组和电针组两组。分别于术后２、４、６、１０周测定运动神经传导速度、小腿三头肌肌力及坐骨神经功能指数，并以自身健侧作对照得出其恢复率。同时测定术后１周感觉神经再生的距离。结果电针组术后１周感觉神经再生距离及２、４、６、１０周运动神经传导速度、肌力、ＳＦＩ等的恢复率均明显优于对照组。结论电针可以促进损伤神经的再生，明显提高神经功能的恢复     

Siatic nerve regeneration in rats stimulated by fibrin glue containing nerve growth factor: an experimental study

Objective

To study the effects of fibrin sealant containing nerve growth factor on the peripheral nerve regeneration.

Study design

A four-group experimental design with repeated measures on one factor was used.

Methods

Fibrin glue (FG) containing NGF was injected into the site of end-and-end sutured peripheral nerve (sciatic nerve) (group I: NGF + FG), meanwhile three control groups were set-up: group II (NGF), group III (FG), and group IV (normal saline). Observation to the function and morphology of the sciatic nerve was carried out at the end of 4, 8, 12 weeks postoperation. Data were analyzed using ANOVA, with the appropriate post hoc between-groups comparison test.

Results

Electrophysiological testing. The NAP and NCV of group I (NGF + FG) were greater than those of group II (NGF), group III (FG), or group IV (normal saline) (p < 0.05). Sciatic functional index (SFI). It began to ameliorate at 4 weeks postoperation and SFI increased as time went on. And the SFI in group I (NGF + FG) was better than those in group II (NGF), group III (FG), or group IV (normal saline) (p < 0.05). Morphology. In the MGF-stained sections, dissociated myelin debris was less and regenerated nerve fibres were in larger quantities in group I (NGF + FG) than in other groups. In the HE-stained sections, regenerated nerve fibres distal to anastomosis significantly increased, and axon and myelin had a clearer outline in group I (NGF + FG) than in other groups. Electron microscopy indicated that the regenerated nerve fibres were more mature and the development of the axons was greater in group I than in other control groups.

Conclusions

FG can be used as carrier of exogenous NGF, and they can provide synergistic effects for the peripheral nerve regeneration after the integration of the two.     

坐骨神经不等径小间隙动脉套接吻合后再生时相的组织学观察

目的 观察大鼠坐骨神经不等径小间隙动脉套接吻合后神经再生的时相变化,探讨其神经再生的机制. 方法 选用Wistar大鼠20只,4只作为套接动脉供体,其余16只切断其右下肢坐骨神经作为实验模型,随机分成坐骨神经等径外膜吻合组和不等径小间隙动脉套接吻合组,每组各8只,分别于术后7、14、21、28 d取材,苏木精-伊红(HE)染色,观察两组神经再生时相的变化. 结果 不等径小间隙动脉套接组再生神经术后21d通过小间隙;手术后28 d,不等径小间隙动脉套接组再生的神经较近端数量增加,再生神经是成熟的,排列规整.结论 近端较细小与远端较粗大的神经通过小间隙动脉套接吻合,神经再生出现放大效应,具有储备功能;小间隙动脉套接吻合是发挥神经再生储备功能的有效手段.     

胶质细胞源性神经营养因子对周围神经再生的作用

目的研究胶质细胞源性神经营养因子(glial cell line-derived neurotrophic factor,GDNF)对周围神经再生的作用.方法硅管套接大鼠坐骨神经,术中一次性将GDNF、生理盐水(nor-mal saline solution,SAL)分别加入硅管中.术后4周,应用溃变神经纤维染色法、辣根过氧化物酶(horseradish peroxidase,HRP)逆行追踪技术观察GDNF对轴突再生的影响.结果与SAL组相比,GDNF组渍变神经纤维面积明显减少,溃变神经纤维面积百分率由17.3%降低到1.9%(P＜0.01);GDNF组脊髓标记胞体显著增加,HRP标记胞体数的百分率由43.5%上升到68.3%(P＜0.01).结论外源性GDNF能明显促进周围神经再生.     

应用Laminin促进周围神经再生的研究

目的探讨局部应用Laminin（LN）对周围神经再生的影响。方法将34只SD大鼠分为4组,1～3组每组10只,手术切除双侧10mm坐骨神经并用硅胶管套接,左侧套管内注入LN0．6μg,右侧注入生理盐水作对照。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逆行示踪检查。结果LN治疗组术后1、3、4个月时的神经肌肉动作电位、运动神经传导速度、再生有髓神经纤维数量及髓鞘厚度均明显优于对照组。相应节段脊髓前角和背根神经节标记神经元明显多于对照组。结论LN可促进周围神经损伤后的再生。