pSVPoMcat微基因修饰雪旺细胞脊髓内移植对操作脊髓再生的影响

Objective In order to observe the role of genetically modified Schwann cell (SC) with pSVPoMcat in the regeneration of injured spinal cord.Method The cells were implanted into the spinal cord.Ninety SD rats were used to establish a model of hemi- transection of spinal cord at the level of T8,and were divided into three groups,randomly, that is,pSVPoMcat modified SC implantation(Group A), SC implantation(Group B),and without cell implantation as control(Group C).After three months the presence of axonal regeneration of the injured spinal cord was examined by means of horseradish peroxidase(HRP)retrograde labeling technique and stereography.Result The results indicated that HRP labeled cells in Group A and B could be found in the superior region of injured spinal cord and the brain stem such as the red nuclei and oculomotor nuclei. The density of ventral horn neurons of the spinal cord and the number of myelinated axons in 100 μ m of the white matter was A >B >C group.Conclusion In brief,the pSVPoMcat modified SC intraspinal implantation could promote regeneration of the injured spinal cord.     

pSVPoMCAT微基因修饰雪旺氏细胞移植治疗实验性脊髓损伤的酶及免疫组化评价

Forthefirsttime，weconstructedmicrogeneforbaseproteinofmyelinsheath（transientlycalledpSVPoMcat）．Usingcationliposome，pSVPoMcatwasinducedintohighlypurifiedschwanncells（SC）．ItwasprovedthatmicrogenepSVPoMcatimplantedtomodifySCcansurviveforalongtimeafterimplantationtoinjuredspinalcord．SCcanpromotemovementfunctionrecoveryafterspinalcordinjury（SCI），butthetherapeuticeffectcannotbeconfirmedmorphologically犤１－２犦．So，theauthorperformedspecificmorphologicalstudyonaxoninthespinalwhi…     

微基因修饰雪旺氏细胞移植对大鼠脊髓损伤后GAP－43表达的影响

Objective To study the effect of microgene pSVPoMcat implanted to modify schwann cell on growth associated protein－43(GAP－43) expression after spinal cord injury in adult rats.Method Hemisected of the T8 segment of the spinal cord was performed for all the experiment rats.The rats were randomly divided into three groups as follows:Group A with microgene pSVPoMcat implanted to genetically modify SC;Group B with SC implanted ;Group C with hemisection of the spinal cord only.The changes of expression of GAP－43 in spinal cord were observed by immunochemistry with antibodies against GAP－43 .Simultaneous,the combined behavioral scores(CBS)was measured.Result There were not any different(P >0.05)among the three groups in first week and 12 week.There were significant diffeence(P<0.05)among three groups in 2nd,8th,and more dxpression of GAP－43 at the 2nd week in group A.The neurofunctional recovery was best in group A.Conclusion The microgene pSVPoMcat implanted to modify schwann cell can promote the expression of GAP－43 in spinal cord and functional recovery in adults rats after SCI.     

pSVPoMCAT微基因修饰雪旺氏细胞移植治疗实验性脊髓损伤的酶及免疫组化评价

Genetherapyofspinalcordinjury（SCI）remainsintheexper－imentalstage．Now，neurotrophicfactortomodifySchwanncell（SC）orfibroblastsareimplantedintoSCIregiontoobservetheregenera－tionsofaxons犤１犦．WefirstbuildPo－５＇－flankinginducedmicrogeneforbasicproteinofmyelinsheathwhichistransientlynamedaspSVPoMcat．ThepSVPoMcatgenewasintroducedintoSCthroughcationliposome．ItwasprovedthatpSVPoMcatcouldenhancefunc－tionofSCandelongateitssurvivalperiodinvitro犤２犦．pSVPoMcatwasimplantedintoS…     

Effect of Microgene pSVPoMcat Implantation to Modify Genetically Schwann Cells on Central Conduction Function after Spinal Cord Injury

We have tested cortical somatasensory evoked potentials (CSEP)after spinal cord injury treated by pSVPoMcat modified Schwann cells implantation and the change of combined behavior score introduced by Gale et al.to explain its effect on central condution after experimental SCI.1 Materials and Methods1.1 SCI model and implantation process Adult SD rats( weighing 200－230g,with no gender regarded,provided by the animal Center of the West China University of medical)were made hemi－transe…     

微基因修饰雪旺氏细胞移植对脊髓损伤中枢传导功能的作用

Objective To approach the effect of microgene pSVPoMcat to modify genetically Schwann cells (SC) on central conduction functionafter spinal cord injury (SCI) . Method Experimental animals were divided into three groups: the group of microgene pSVPoMcat implanted to genetically modify SC (group A), SC implanted group (group B), and the control group (group C). The cortical somatasensory evoked potentials (CESP) and combined behavioral score (CBS) were continuous monitored from operation second week to twelfth week Result The result showed that the peak latency and peak amplitudes of group A, B have a recovery tendency and it was constant with CBS. Conclusion We concluded that the microgene pSVPoMcat to modify genetically Sc may play a promotion role in recovery of central conduction function after SCI.     

pSVPoMcat微基因修饰雪旺细胞脊髓内移植对损伤脊髓细胞的保护作用

Objective To study the protective effects of the intracord transplantation of microgene pSVPoMcat－ genetically－ modified Schwann cells (MSCs)on spinal cord injury (SCI).Method Rats with semi－ division(SD) of the spinal cord was divided into 4 groups.Group S consisted of the rats with SD treated with the transplantation of MSCs, Group B of the rats with SD treated with the transplantation of SCs without genetic modification,Group C of the rats with SD without treatment and Group D was the normal control. 8 hours after operation,the half of the rats of each group were killed and the injured segment of the spinal cord was resected to be examined with atomic absorption spectrophotometry . Another half of the rats of all the groups were examined with neurological function tests to have a combined behavioral score (CBS).Result There was a significant increase of water content and Na＋ and Ca2＋ ions and a decrease of K＋ and Mg 2＋ ions in the injured cord segment of Group C and a statistically significant recovery was observed in Group A. The intracord transplantation of pSVPoMcat genetically modidied SCs improved the neurological outcome of spinal cord injury.Conclusion Our findings indicate that intracord transplantation of pSVPoMcat－ genetically－ modified－ Schwanncells exerts protective effects on the injured segment of the spinal cord through the improvement of the internal ion environment of the spinal cord.     

The effect of microgene pSVPoMcat to modify Schwann cell on GAP -43 expression after spinal cord injury in adult rats

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人神经生长因子基因修饰的许旺细胞治疗脊髓损伤的实验研究

目的:观察神经生长因子(nerve growth factor,NGF)基因修饰的许旺细胞(Schwann cells,SCs)对大鼠脊髓损伤后运动功能恢复及GAP-43的表达情况,探讨其对脊髓损伤修复的作用机制.方法:清洁级SD大鼠52只,随机分为EGFP-N1-NGF转染许旺移植组16只(A组),许旺细胞移植组16只(B组),PBS对照组12只(C组)和假手术组8只(D组),用改良Allen法制造大鼠脊髓损伤模型.分别于术后7,14,21,28 d用BBB评分法观察动物行为学的变化,用肌电图检测动物中枢传导速度和波幅;然后处死动物,取损伤区1 cm范围脊髓节段,常规石蜡包埋,组织切片,GAP-43免疫组织染色观察脊髓损伤的组织学变化.结果:BBB评分结果D组＞A组＞B组＞C组;肌电图检测结果显示中枢传导速度A、B、C组间差别有统计学意义P＜0.05,D组明显高于其它三组;GAP-43免疫组化检测结果提示A＞B＞C组,D组仅有少量表达.结论:本研究证实NGF基因修饰的许旺细胞能够促进脊髓损伤的恢复,可能是促进神经轴突的生长有关.     

延迟植入嗅鞘细胞修复成鼠的胸髓损伤

背景:脊髓损伤后在一定的微环境中有再生的能力。嗅鞘细胞兼具星形胶质细胞和许旺细胞的特性,具有促进脊髓轴突再生的能力。目的:制作成鼠胸髓损伤模型,观察嗅鞘细胞对损伤脊髓轴突再生的作用。设计:观察性实验。单位:中山大学附属第二医院。材料:实验于2001-01/2002-11在中山大学附属第二医院完成。20只成年SD雄性大鼠,体质量(380±20)g,由中山大学实验动物中心提供(机构许可证号:SYXK(粤)2004-0020)。低糖的DMEM培养液(L-DMEM,GibcoBRL公司)、胎牛血清(Hyclone公司)、MBP(髓磷脂碱性蛋白,Sigma公司)、抗神经生长因子受体抗体(Sigma公司)。应用随机数字表完全随机化分为细胞移植组和对照组,每组10只。方法:将成年SD大鼠麻醉断颈处死,无菌条件下取出完整嗅球分嗅神经。采用改良Allen法打击脊髓建立胸髓损伤模型。细胞移植组于损伤脊髓处注入10μL嗅鞘细胞悬液(2.5×1010L-1),对照组仅注入相同剂量DMEM/F12(1∶1)培养液。移植后6周通过组织学和免疫组织化学方法观察嗅鞘细胞对脊髓轴突再生的影响。主要观察指标:①抗神经生长因子受体抗体染色鉴定嗅鞘细胞。②髓磷脂碱性蛋白染色观察髓鞘的修复。③嗜银染色观察神经轴突再生。结果:细胞移植组有2只动物死亡,对照组有3只死亡,共15只大鼠进入结果分析。①细胞移植组可见损伤脊膜完整,较正常脊髓稍变细。苏木精-伊红染色见损伤区有多极细胞,且与脊髓组织有移行过度现象,说明存活的嗅鞘细胞与宿主融合良好。新生的轴突多呈束状,周围有小圆淋巴细胞浸润。嗜银染色可见再生轴突长入损伤区组织中,多与束状排列的多极细胞伴行;对照组脊髓损伤处明显变细,脊膜尚完整。苏木精-伊红染色见脊髓损伤区未见新生轴突。嗜银染色未见再生轴突。②细胞移植组可见多极细胞,多呈束状排列,胞浆内有大量抗神经生长因子受体抗体阳性颗粒存在,进一步证明嗅鞘细胞移植6周后仍然存活,且与宿主融合良好。髓磷脂碱性蛋白染色见损伤区有呈线状髓磷脂碱性蛋白阳性纤维,提示损伤区两端均有髓鞘样物质产生,且互相靠拢。同时多极细胞中也发现有髓磷脂碱性蛋白阳性物质存在,说明移植的嗅鞘细胞有产生髓鞘样物质的作用。对照组未见轴突再生。结论:嗅鞘细胞延迟植入成鼠打击伤后脊髓后,可存活并产生髓鞘样物质,促进宿主脊髓轴突再生。