嗅鞘细胞移植对大鼠实验性自身免疫性脑脊髓炎的影响

目的:探索嗅鞘细胞移植实验性自身免疫性脑脊髓炎(EAE)大鼠的移植途径、可行的移植时间窗,移植后的迁移特性以及发挥保护作用的可能机制。方法:分别采用豚鼠脊髓匀浆(GPSCH)与MOGIgd融合蛋白免疫Lewis大鼠,制作EAE模型;每组发病大鼠分别归入:MOG组和GPSCH组。MOG组分为:OECs空白对照组(MOG0组)4只、OECs尾静脉移植组(MOG1组)7只、OECs侧脑室移植组(MOG2组)4只;GPSCH组分为:OECs空白对照组(GPSCH0组)4只、OECs尾静脉移植组(GPSCH1组)4只。发病高峰期,按照实验分组,分别采用立体定向侧脑室细胞移植和尾静脉细胞移植,观察移植后大鼠的临床症状;移植后2周观察OECs在大鼠体内的分布情况及组织病理学方面的缓解情况。结果:OECs分别经尾静脉、侧脑室移植后,EAE大鼠症状改善,与空白对照组神经功能评分峰差值比较有显著差异(F=18.470,P0.01;t=-7.147,P0.01),MOG1组和MOG2组评分峰差值间无显著差异(P0.05);Hoechst33342示踪证实了OECs能在大鼠体内存活及强大的迁移能力;OECs经尾静脉移植后,可以透过破坏的血脑屏障进入脑内,分布于软脑膜和病灶周围;经侧脑室移植的嗅鞘细胞,向病灶局部广泛迁移;组织病理学评分(HE染色和Luxol fast blue髓鞘染色),移植组与未移植组间无显著差异(P0.05),2种途径移植组间亦无显著差异(P0.05)。结论:纯化培养的成年大鼠嗅球嗅鞘细胞分别经尾静脉和侧脑室移植EAE大鼠,均可缓解发病大鼠的症状。     

Olfactory Ensheathing Cell Transplantation Into Spinal Cord Prolongs the Survival of Mutant SOD1G93A ALS Rats Through Neuroprotection and Remyelination

Amyotrophic lateral sclerosis (ALS) is a progressively fatal, incurable, neurodegenerative disorder. In this study, we investigated whether olfactory ensheathing cells (OEC) transplantation could provide protection to motor neurons and enable remyelination in mutant SOD1^G93A transgenic rats with ALS. Seventy‐two rats were divided into four groups: SOD1^G93A rats (n = 20); medium+SOD1^G93A rats (n = 20); OECs+SOD1^G93A rats (n = 24); and another eight wild‐type rats were used as controls. About 5 μL (1 × 10⁵) OECs in DF12 medium was injected into the dorsal funiculus of the thoracic spinal cord at a predetermined depth. Survival analysis revealed a significant increase in the survival time in OEC+SOD1^G93A rats. Body weight records and inclined board test showed a significant difference between OEC+SOD1^G93A and SOD1^G93A from the onset at 7 days to 11 days (P < 0.05). Four weeks following transplantation, motor neuron counts in the ventral horn of the spinal cord noted a significant motor neuron loss in SOD1^G93A rats when compared with wild‐type rats (P < 0.001), and much less neuronal loss and collapse was noted in OEC+SOD1^G93A rats when compared with SOD1^G93A rats(P < 0.001); immunohistochemistry and Western blot analysis of choline acetyltransferase supported the motor neuron count. Images of confocal microscope indicated that the transplanted OECs had survived for more than 4 weeks and migrated 4.2 mm through the spinal cord. Evidence of remyelination of transplanted OEC was captured with triple fluorescence labeling of green fluorescent protein, neurofilament, and myelin basic protein and was further confirmed by Western blot analysis of MPB. In conclusion, the transplanted OECs could serve as a source of neuroprotection and remyelination to modify the ALS microenvironment. Anat Rec, 2011. © 2011 Wiley‐Liss, Inc.     

实验性变态反应性脑脊髓炎大鼠嗅鞘细胞移植排斥反应研究

目的研究嗅鞘细胞(OECs)的免疫原性及移植实验性免疫性脑脊髓炎(EAE)后颈淋巴系统的反应。方法用新生ICR小鼠嗅球培养出OECs,流式细胞术(FCM)检测其表面主要组织相容性抗原I、II类(MHC-I、MHC-II)诱导型和组成型表达情况。将经荧光染料CFSE标记OECs悬液或PBS注入EAE发病大鼠,收集注射7d后颈部淋细胞,FCM检测CFSE+、抗原提呈细胞标志物CD83+及CFSE+CD83+的表达率,混合淋巴细胞培养检测淋巴细胞增殖率(PI)。结果 OECs组成型表达少量MHC-I分子但几乎不表达MHC-II分子,但能在IFN-γ作用下能显著上调这二类分子。移植实验显示移植OECs的EAE大鼠颈部淋巴结中CD83+、CFSE+及CD83+CFSE+检出率及OECs诱导的PI值均高于对照组或正常大鼠(P<0.05)。同时,PBS注射的EAE大鼠,较正常大鼠亦有较高的CD83+检出率及PI值(P<0.05)。结论 OECs组成型或诱导型表达的MHC分子提示OECs有一定免疫原性,移植后局部淋巴系统可出现针对OECs的抗原引流、提呈及T细胞的活化,进一步分析提示这种免疫反应的启动依赖于EAE的炎性环境。     

The Immunohistochemical Characterization of Human Fetal Olfactory Bulb and Olfactory Ensheathing Cells in Culture as a Source for Clinical CNS Restoration

Clinical studies have expanded the therapeutic olfactory ensheathing cells (OECs) transplantation to different human Central Nervous System (CNS) diseases. In fact, the OEC transplantation in clinic is a mixture of olfactory bulb cells; they even have not demonstrated that they have such a subpopulation yet. However, as a source of OECs transplantation, the development and identification of human fetal OECs are still need more understanding, because some surgery try to restoration CNS injury with a more purity of OEC cultures generated by a number of different procedures. In this article, twelve human fetal olfactory bulb (OB) samples were obtained from six fetuses in 20 weeks of gestation, it was studied by immunofluorescence on histological sections and cultured cells with multiple antibodies under confocal microscopy. The P75NTR positive OB‐OECs (olfactory ensheathing cell from the olfactory bulb) were present in both outer olfactory nerve layers and glomerular layer. The percentage of OB cells in culture, about 22.31 was P75NTR positive, 45.77 was S100β, and 31.92 was GFAP. P75NTR and GFAP were coexpressed with S100β, respectively; however, P75NTR was not coexpressed with GFAP in human fetal OECs. It is suggested that the localization and development of human OECs in OB are different to those in rodent, and the P75NTR immunohistological staining is still necessary to identify and characterize human fetal OECs in culture before transplantation. Anat Rec, 2010. © 2009 Wiley‐Liss, Inc.     

嗅鞘细胞联合神经干细胞脑内移植改善Parkinson病大鼠行为的观察

为了观察嗅鞘细胞(OECs)联合神经干细胞(NSCs)脑内移植对Parkinson病(PD)的治疗作用,首先将6-OHDA注射至左侧黑质致密部和腹侧被盖区制备PD大鼠。将成功制备的PD大鼠随机分为5组,即0.9%生理盐水对照组、单纯移植NSCs或OECs组、OECs+NSCs共移植组和PD模型对照组。将培养并传至第3代的NSCs、OECs和OECs+NSCs分别移植到PD大鼠的纹状体内,于移植后7、14、30、60d检测PD大鼠旋转行为变化后处死,取脑做冰冻切片,行BrdU、酪氨酸羟化酶(TH)、p75免疫荧光和TH免疫组织化学检测。移植后21d以内,各组大鼠的旋转行为无明显变化;30d和60d时,单纯移植NSCs或OECs组和OECs+NSCs共移植组与生理盐水和模型对照组相比,旋转行为有较明显的改善(P<0.05),其中OECs+NSCs共移植组的旋转行为改善更为显著。在移植后30d和60d,单纯移植NSCs或OECs组在移植区可见少数TH阳性神经元,而OECs+NSCs共移植组的TH阳性神经元明显多于单纯移植NSCs或OECs组(P<0.05)。上述结果表明:OECs能促进胚鼠中脑来源的NSCs向TH阳性神经元分化;单纯移植NSCs或OECs和OECs+NSCs共移植,均能不同程度地改善PD大鼠的旋转行为,但联合移植优于单纯移植。     

嗅鞘细胞和神经干细胞联合移植阿尔茨海默病大鼠脑内的增殖和定向分化

背景：阿尔茨海默病大鼠脑内神经元减少,神经干细胞移植后增殖和向神经元分化能力有限。 目的：观察联合移植嗅鞘细胞和神经干细胞在阿尔茨海默病大鼠脑内,嗅鞘细胞对神经干细胞的增殖和向胆碱能神经元分化的影响。 方法：体外培养嗅鞘细胞和神经干细胞,移植前用5-溴脱氧尿嘧啶核苷标记神经干细胞。将生理盐水,神经干细胞和神经干细胞+嗅鞘细胞分别移植入阿尔茨海默病模型大鼠海马。移植7,14,21,28 d后,进行大鼠脑组织切片免疫组织化学染色检测BrdU和ChAT阳性表达。 结果与结论：联合移植组神经干细胞的增殖和分化情况明显优于其他两组,联合移植组BrdU阳性细胞数和ChAT阳性细胞数明显高于神经干细胞移植组和生理盐水组(P < 0.01)。提示嗅鞘细胞能促进移植的神经干细胞在阿尔茨海默病大鼠脑内增殖和向胆碱能神经元的分化。     

尾静脉移植羊膜间充质干细胞治疗大鼠脑缺血再灌注损伤

背景：羊膜间充质干细胞是一种理想的再生医学领域的种子细胞来源。 目的：探讨尾静脉移植羊膜间充质干细胞对脑缺血损伤的作用。 方法：线栓法制备大鼠大脑中动脉梗死模型,缺血2 h后进行再灌注。造模后24 h通过尾静脉移植1.0×105个人羊膜间充质干细胞,于造模后2,4周完成神经功能评分后取材。以未移植的模型大鼠为对照。 结果与结论：脑损伤后,大鼠出现神经功能评分明显增高,羊膜间充质干细胞移植1周,大鼠的神经功能评分显著降低(P < 0.05),移植2,4周时,神经功能进一步改善。免疫荧光染色显示,移植羊膜间充质干细胞的大鼠脑组织中可见较多发绿色荧光的BrdU阳性细胞,主要位于缺血区周围、皮质下和侧脑室。说明尾静脉移植羊膜间充质干细胞可迁移至大鼠脑缺血区,改善大鼠的神经功能。     

侧脑室移植骨髓基质细胞对脑缺血再灌注大鼠运动及认知功能的影响

背景：将骨髓基质细胞经侧脑室移植治疗大脑中动脉阻断缺血所致脑梗死模型鼠已取得一定效果,但尚未见对其认知功能的影响。 目的：观察骨髓基质细胞侧脑室移植对大脑中动脉阻断缺血模型大鼠行为认知功能的影响,并观察脑梗死灶大小的变化和植入骨髓基质细胞的迁移路径。 方法：制作SD大鼠大脑中动脉阻断缺血2 h再灌注模型后随机分为3组,骨髓基质细胞组及磷酸缓冲液组分别于梗死侧侧脑室注射骨髓基质细胞悬液5 μL(含约1.0×10⁶个细胞)或等量的磷酸缓冲液,模型组和不造模的正常组不作任何处理。应用平衡木实验观察大鼠运动协调能力,水迷宫实验观察其游泳速度及空间学习记忆能力;苏木精-伊红染色观察梗死灶大小的变化,免疫组化观察BrdU阳性细胞的迁移路径。 结果与结论：骨髓基质细胞组在移植后第3,7,14天平衡木评分均有显著改善(P < 0.05)。移植后第7~10天,骨髓基质细胞组大鼠的游泳速度均快于磷酸缓冲液组(P  < 0.05)。骨髓基质细胞组大鼠寻找平台潜伏期的时间明显缩短(P < 0.05),在原平台象限所占时间百分比和路程百分比及穿越平台次数明显增加(P < 0.05)。移植后7,14 d,各模型组大鼠苏木精-伊红染色均可见典型的脑缺血梗死病灶,梗死面积百分比无明显差异。BrdU染色显示,移植后第1天,阳性细胞都聚集在移植侧侧脑室,以紧密排列的细胞团形式存在于脑室壁;第3天大部分细胞穿越脑室壁以单个细胞形式向周围缺血区迁移;第14天移植细胞在梗死的纹状体、皮质可见。提示大脑中动脉阻断缺血后24 h侧脑室移植骨髓基质细胞可明显改善运动协调能力及空间学习记忆能力;移植骨髓基质细胞未能减小梗死灶大小,但侧脑室移植的骨髓基质细胞可存活并定向性地迁移至缺血的纹状体和皮质。     

嗅鞘细胞移植联合督脉电针治疗大鼠前脊髓综合征

背景：电针在脊髓损伤的康复过程中有其一定的临床作用,同时以嗅鞘细胞为代表的细胞治疗在部分患者的康复过程中亦起到了一定的作用,两者是否具有协同应用尚不清楚。 目的：观察督脉电针联合嗅鞘细胞移植对前脊髓综合征大鼠神经营养因子3水平及P75NTR表达的影响。 方法：将40只成年大鼠随机分为对照组、督脉电针组、嗅鞘细胞移植组、督脉电针+嗅鞘细胞移植组,经嗅鞘细胞移植和督脉电针治疗2周后 ELISA法检测脊髓组织神经营养因子3水平,免疫组化检测P75NTR的表达。 结果与结论：督脉电针+嗅鞘细胞移植组神经营养因子3水平较对照组及其他实验组高(P < 0.05);嗅鞘细胞移植组和督脉电针+嗅鞘细胞移植组的脊髓损伤部位以及相邻的组织内均有P75NTR表达,且督脉电针+嗅鞘细胞移植组阳性表达明显较嗅鞘细胞移植组多。实验证实督脉电针联合嗅鞘细胞移植能够明显增高大鼠前脊髓综合征邻近组织的神经营养因子3水平;督脉电针可以有效促进移植的嗅鞘细胞在宿主内存活。     

Therapeutic potential of olfactory ensheathing cells in neurodegenerative diseases

The regenerative capacity of the olfactory system has generated interest in potential clinical application of cells from the olfactory epithelium in the treatment of neurodegenerative diseases. Experimental evidence from animal models and clinical studies suggest that transplantation of olfactory ensheathing cells (OEC), specialized glia in the olfactory system, may be therapeutically useful in neurodegenerative diseases such as spinal cord injury and stroke. This review article describes the different experimental approaches in OEC transplantation. We also discuss the possible effects of OEC implantation on the underlying pathophysiology in neurological disease, including neuroplasticity. Our recent study of this particular population of cells has disclosed some of the molecular basis of the regenerative mechanism of OECs. In summary OECs produce several neurotrophic factors such as stromal cell-derived factor 1α and brain-derived neurotrophic factor and enhance axonal regeneration to promote neuroplasticity in neurodegenerative diseases.     

In vivo MRI of olfactory ensheathing cell grafts and regenerating axons in transplant mediated repair of the adult rat optic nerve

The purpose of the present study was to use magnetic resonance imaging (MRI) as a tool for monitoring transplant-mediated repair of the adult rat visual pathway. We labelled rat olfactory ensheathing cells (OECs) using micron-sized particles of iron oxide (MPIO) and transplanted them by: i) intravitreal injection (ivit) and ii) intra-optic nerve (ON) injection (iON) in adult rats with ON crush (ONC) injury. We applied T(2)-weighted MRI and manganese-enhanced MRI (MEMRI) to visualise transplanted cells and ON axons at specific times after injury and cell engraftment. Our findings demonstrate that ivit MPIO-labelled OECs are unequivocally detected by T(2)-weighted MRI in vivo and that the T(1)-weighted 3D FLASH sequence applied for MEMRI facilitates simultaneous visualisation of Mn(2+-) enhanced regenerating retinal ganglion cell (RGC) axons and MPIO-labelled OEC grafts. Furthermore, analysis of MRI data and ultrastructural findings supports the hypothesis that iON OEC transplants mediate regeneration and remyelination of RGC axons post injury.     

骨髓间充质干细胞静脉移植治疗大鼠脊髓损伤

背景：干细胞移植可以重建中枢神经系统的结构和功能,近年来引起了广泛的关注。 目的：尾静脉移植骨髓间充质干细胞观察其对大鼠脊髓损伤修复的影响并探讨其机制。 方法：密度梯度离心法结合贴壁筛选法分离骨髓间充质干细胞。将成年雌性Wistar大鼠以动脉瘤夹钳夹法制备大鼠脊髓损伤模型后随机分为细胞移植组和对照组,分别干预。 结果与结论：细胞移植组大鼠BBB后肢功能评分恢复优于对照组(P < 0.05)。自损伤后第30天开始,细胞移植组大鼠运动诱发电位潜伏期及体感诱发电位P1波潜伏期的恢复均优于对照组(P < 0.05),并持续至实验结束。细胞移植组大鼠脊髓内在损伤中心及头、尾端距离脊髓损伤中心1 cm处均可见阳性细胞。说明尾静脉注射移植骨髓间充质干细胞可以促进脊髓损伤大鼠神经功能恢复,其机制可能与移植细胞迁移到损伤部位,分化为神经元样和神经胶质细胞样细胞,并分泌或促进宿主分泌神经营养因子有关。     

人骨髓间质干细胞和嗅鞘细胞移植对大鼠急性脊髓损伤功能修复的比较研究

目的对比人骨髓基质细胞(BMSCs)与人胚嗅鞘细胞(OECs)移植对大鼠脊髓损伤功能修复的影响。方法将45只SD大鼠分成脊髓损伤后BMSCs移植组(BMSCs组)、OECs移植组(OECs组)和PBS对照组(PBS组),通过BBB评分、运动诱发电位(MEP)评估脊髓传导功能的改善状况,免疫组织化学方法检测移植细胞存活和分化情况,病理形态学方法观察组织结构修复情况。结果BMSCs组BBB评分高于OECs组(P<0.05);两细胞治疗组MEP潜伏期明显缩短(P<0.05);BMSCs组嗜银染色可见脊髓损伤近端有较多再生纤维向远端延伸,形成神经纤维束,而OECs组再生纤维较少;两种移植细胞均可在损伤处部分存活,BMSCs组可见BMSCs来源的细胞Nestin、NF、GFAP的阳性表达。结论BMSCs移植比OECs移植能更有效促进大鼠急性脊髓损伤修复。     

自组装肽包裹嗅神经鞘细胞抑制脊髓损伤后胶质瘢痕形成并促进轴突再生

目的研究自组装肽包裹嗅神经鞘细胞(OECs)移植对大鼠脊髓损伤(SCI)后轴突再生的影响。方法成年雄性SD大鼠40只,行脊髓T_(10)节段背侧横断术,动物随机分为4组,每组10只。SAP-OECs组于间隙内移植SAP包裹的OECs悬液,SAp、OECs对照组同法分别移植等量SAP及DF12-OECs悬液;单纯全切对照组不做处理。术后2或4周过量麻醉处死动物,脊髓损伤节段连续冰冻矢状切片。行抗GAP-43和GFAP免疫组织化学和免疫荧光双标染色。结果SAP-0ECs组损伤区内有大量GAP-43免疫反应纤维,SAP组相较OECs及单纯半切对照组则可见相当数量GAP-43免疫反应纤维。OECs及单纯半切对照组损伤区周围可见一条致密度的GFAP阳性的反应性星形胶质细胞条带,而SAP组GFAP免疫反应纤维比对照组少,且多于SAP-OECs组。此外激光共聚焦显微镜下观察SAP-OECs组在损伤后4周GAP-43免疫反应纤维能通过位于脊髓损伤区尾端的GFAP阳性区域。结论自组装肽包裹嗅鞘细胞可以抑制脊髓损伤后胶质疤痕形成并促进轴突再生。     

新生大鼠神经干细胞移植对脑缺血再灌注损伤的治疗作用

目的:探讨新生大鼠神经干细胞移植治疗局灶性脑缺血再灌注损伤的可行性及疗效.方法:体外培养新生大鼠神经干细胞.采用改良的线栓法制作脑缺血再灌注模型,3d后用脑立体定位仪经脑室移植神经干细胞,移植时间点及再灌注1～7周对移植大鼠进行神经功能损伤程度评分(NSS).再灌注1、 2、 3、 5、 7周末麻醉处死大鼠,脑组织石蜡包埋.免疫组织化学方法观察移植后神经干细胞的存活、分布.结果:神经干细胞表达巢蛋白,在血清条件下分化为表达微管相关蛋白(MAP2)的神经元和表达胶质细胞纤维酸性蛋白(GFAP)的星形胶质细胞.神经干细胞移植组NSS评分在各个时间点均显著低于对照组.移植的神经干细胞分布于缺血侧皮质、纹状体,再灌注后3、 5、 7周,皮质、纹状体阳性细胞数分别较1、 2周显著增多,第3、 5、 7周之间差异无统计学意义.前3周组织结构疏松,缺损严重,而第5、 7周组织结构较前3周完整致密.结论:移植的神经干细胞能在脑缺血大鼠脑内存活、迁移,并能改善缺血后大鼠的神经功能状况.     

嗅鞘细胞静脉移植治疗脊髓损伤

背景：嗅鞘细胞移植治疗脊髓损伤在众多疗法中效果较佳,成为最有前景的治疗方法之一。目前移植方法为局部移植,存在操作复杂、创伤大、重复移植治疗困难等缺点。寻找一种简单易行且疗效好的移植方法成为各国学者研究的热点。 目的：分析嗅鞘细胞静脉移植治疗脊髓损伤的可行性和疗效。 方法：制备Wistar大鼠脊髓半切模型,随机分4组：嗅鞘细胞髓内局部移植组、嗅鞘细胞静脉移植组、D/F12静脉移植组和空白对照组。定期行CBS功能评分及组织学检查,评价脊髓修复情况。 结果与结论：嗅鞘细胞髓内局部移植组、嗅鞘细胞静脉移植组的功能恢复和组织学改变优于D/F12静脉移植组和空白对照组,嗅鞘细胞髓内局部移植组、嗅鞘细胞静脉移植组间无显著差别。说明嗅鞘细胞静脉移植可向脊髓损伤部位迁移并修复脊髓损伤,其疗效与嗅鞘细胞髓内局部移植相当。     

Neuropathology of Olfactory Ensheathing Cell Transplantation into the Brain of Two Amyotrophic Lateral Sclerosis (ALS) Patients

Although a large number of amyotrophic lateral sclerosis (ALS) patients have undergone transplantation procedures with olfactory ensheathing cells (OECs) in the Bejing Hospital, to our knowledge, no post‐mortem neuropathologic analyses have been performed. We examined the post‐mortem brain of two Italian patients affected by ALS who underwent cellular transplantation in Beijing with their consent. Our aim was to assess the events following the graft procedure to possibly support the rationale of the treatment strategy. The neuropathologic findings were analyzed on the basis of the limited awareness of the experimental conditions and discussed in relation to the safety, efficacy and long‐term outcome of the transplanted cells. Islands of quiescent, undifferentiated cells within the delivery track persisting for up to 12 months–24 months were found. Prominent glial and inflammatory reaction around the delivery track strongly supports the encasement of the graft. Evidence of axonal regeneration, neuronal differentiation and myelination was not seen. The surgical procedure of implantation was not compatible with a neurotrophic effect. The OEC transplantation did not modify the neuropathology of ALS in the two patients. In conclusion, the present neuropathologic analysis does not support a beneficial effect of fetal OEC implantation into the frontal lobes of ALS patients.     

嗅鞘细胞移植脊髓损伤大鼠Sema3A及其受体NP-1基因的表达

背景：嗅鞘细胞移植促进脊髓损伤修复机制中对神经生长抑制导向因子的研究较少。 目的：观察嗅鞘细胞移植对脊髓半横断损伤后神经生长抑制导向因子Sema3A及其受体NP-1在RNA水平表达的变化。 方法：17只SD大鼠随机分为嗅鞘细胞移植组、DMEM/F12 培养液移植组和正常对照组。制作T11~T12水平大鼠脊髓左侧半横断损伤模型,立刻分别注射嗅鞘细胞悬液或等量的DMEM/F12 培养液。 结果与结论：6周后取横断水平前后两个脊髓节段,用RT-PCR的方法对Sema3A、NP-1的mRNA进行半定量分析。①嗅鞘细胞在大鼠脊髓内仍有大量存活。②Sema3A及NP-1 mRNA的量,DMEM/F12 培养液移植组明显多于正常对照组(P < 0.05),嗅鞘细胞移植组与培养液移植组比较明显下调,且与正常对照组比较差异无显著性意义(P > 0.05)。③提示嗅鞘细胞移植有效减少了神经生长抑制导向因子Sema3A及其受体NP-1 mRNA的表达。     

嗅鞘细胞移植对脊髓半横断损伤大鼠后肢运动功能的影响

目的观察嗅鞘细胞移植物对脊髓半横断损伤大鼠后肢运动功能的影响。方法30只雌性成年SD大鼠制成脊髓半横断损伤模型,并随机分成5组立即进行移植,A组移植纯化的嗅鞘细胞(olfactory ensheathing cells,OECs)悬液,B组移植纯化的OECs培养上清液,C组移植未纯化的OECs,D组移植未纯化的OECs培养上清液,E组移植DMEM/F12培养液,术后6 w内进行BBB评分和IP实验,用单因素方差分析进行统计学分析。结果A组的BBB评分在3w后较其它组高,差异有统计学意义(P<0.05),A组的IP实验结果在4 w后较其它组高,差异有统计学意义(P<0.05)。结论纯化培养嗅鞘细胞移植能促进脊髓半横断损伤大鼠后肢运动功能恢复。     

嗅鞘细胞移植对大鼠脊髓半横断损伤后轴突再生及后肢功能恢复的作用

为了对嗅鞘细胞移植治疗大鼠脊髓损伤(spinal cord injury, SCI)后轴突再生及后肢功能恢复进行综合性的评价,本实验采用22只雌性成年SD大鼠,并随机分成A、B、C、D 4组。其中A、B、D组行左侧T11 ～T12节段脊髓半横断手术,然后A组移植嗅鞘细胞(olfactory ensheathing cells,OECs)悬液;B组移植DMEM/F12培养液;D组移植核荧光试剂(Hoechst33342)标记的嗅鞘细胞,用于鉴定嗅鞘细胞在体内的存活情况;C组作为正常对照组。术后6周内,对大鼠进行BBB评分、IP斜板试验并观察皮质体感诱发电位(cortical somatosensory evoked potential, CSEP)、运动诱发电位(motor evoked potential, MEP)的潜伏期。将辣根过氧化物酶(HRP)注射到横断面尾段脊髓,逆行追踪观察横断面头段脊髓及对侧中脑红核内HRP逆标细胞数,并观察左侧后肢小腿三头肌肌细胞横截面积及直径的变化。结果显示:(1)移植后的OECs数量未见明显减少,细胞核形态较好,细胞未向头、尾侧迁移;(2)移植3周后BBB评分及IP斜板,试验A、B组较C组明显低(P<0.05),但A组明显高于B组(P<0.05);(3)A、B组的CSEP及MEP潜伏期较C组明显延长(P<0.05),但A组要比B组明显缩短(P<0.05);(4)HRP逆行追踪观察到对侧中脑红核大细胞区及脊髓T9 ～T11节段逆标细胞的数量,A组明显多于B组,但A、B组均少于C组;(5)左侧后肢小腿三头肌肌细胞的横截面积及直径,A、B组均较C组减少,但A组减小的幅度明显小于B组。以上结果表明OECs移植能促进半横断脊髓轴突的再生及后肢功能的恢复。