大鼠脑缺血再灌注损伤后细胞凋亡与Survivin和IGF-1表达的关系

目的 探讨大鼠脑缺血再灌注损伤后Survivin和IGF-1因子表达与细胞凋亡的关系.方法 将80只成年健康雄性Wistar大鼠,分为Survivin组和IGF-1组,每组各40只,并随机分为对照组、假手术组和缺血1h再灌注2h、6h、12h、24h、48h、3d、7d、14d组,每组4只(n=4).应用线栓法制备大鼠脑缺血再灌注模型(MCAO),采用免疫组织化学方法检测Survivin蛋白与IGF-1蛋白在神经元凋亡中的表达情况.结果 Survivin组:假手术组Survivin阳性神经元在海马、皮质区及纹状体区呈基础表达,缺血再灌注2h表达明显增加,48h达高峰,14d表达最低,与假手术组比较有显著性差异(P＜0.05).IGF-1组:正常对照组及假手术组IGF-1阳性神经元在海马、皮质区及纹状体区呈基础表达,缺血再灌2h时IGF-1表达明显增高,48h达高峰,3～14d仍维持高表达,与正常对照组及假手术组比较有显著性差异(p＜0.05).结论 内源性Survivin与IGF-1因子可能具有在抑制神经元凋亡的作用.     

骨髓基质细胞移植缺血性脑梗死大鼠后神经营养因子的表达

背景：骨髓基质细胞在适宜条件下可分化为神经元和星形胶质细胞,分泌可溶性分子促进神经元存活。 目的：观察骨髓基质细胞移植后缺血性脑梗死大鼠脑组织神经营养因子表达情况及其对神经功能的影响。 方法：改良的Longa栓线法制作大鼠大脑中动脉缺血模型,1 h后再灌注,24 h后治疗组尾静脉注射3×106骨髓基质细胞,盐水对照组尾静脉注射1 mL生理盐水,空白对照组不进行注射。 结果与结论：在梗死后第7,14天治疗组的神经损伤评分明显低于对照组(P < 0.05);治疗组神经生长因子和脑源性神经营养因子表达在各时间点均高于对照组(P < 0.05)。结果显示静脉移植骨髓基质细胞可改善缺血性脑梗死大鼠神经功能,移植骨髓基质细胞后缺血脑组织中神经生长因子及脑源性神经营养因子表达促进了神经功能的恢复。     

骨髓基质细胞静脉移植治疗大鼠短暂性局灶性脑缺血

目的探讨骨髓基质细胞静脉移植治疗大鼠短暂性局灶性脑缺血的可行性及其机制。方法将大鼠骨髓基质细胞在体外纯化、扩增并经BrdU标记后,经尾静脉移植到局灶性脑缺血大鼠体内,通过神经缺损评分观察移植后大鼠神经行为学改善情况,通过组织学方法观察移植到脑内的骨髓基质细胞表达脑源性神经营养因子、缺血灶周围细胞凋亡及脑微血管密度的变化。结果骨髓基质细胞移植组大鼠的神经缺损评分显著低于对照组(P<0.05);移植到脑内的骨髓基质细胞主要选择性分布于缺血灶周围区域并表达BDNF;骨髓基质细胞移植组大鼠梗死灶周围的凋亡细胞明显少于对照组(P<0.01)、微血管密度显著高于对照组(P<0.001)。结论经静脉注射移植骨髓基质细胞能够明显促进局灶性脑缺血大鼠的神经行为功能恢复;抗凋亡及促微血管增生可能是骨髓基质细胞移植治疗局灶性脑缺血的机制之一。     

人脐血细胞静脉移植对自发性高血压大鼠缺血再灌注脑损伤疗效及其机制探讨

目的探讨人脐血细胞(HUCBCs)静脉移植治疗自发性高血压(SHR)大鼠缺血再灌注脑损伤疗效及其相关机制。方法人脐血细胞体外分离、扩增并以4,6-2胺-2苯基吲哚(DAPi)标记。68只雄性SHR大鼠经线栓法建立大鼠大脑中动脉栓塞(MCAO)模型,并随机分为2组:治疗组和对照组。术后48h后经尾静脉治疗组注射1×106HUCBCs,对照组注射HUCBCs细胞培养基。于移植后3、14、28d,Bederson评分观察大鼠神经功能状况;图像分析法检测计算各组TTC染色脑梗死面积;荧光显微镜对治疗组脑组织DAPi染色阳性细胞计数;免疫组化法检测脑组织内脑源性神经营养因子(BDNF)染色阳性细胞数以及逆转录聚合链式反应(RT-PCR)检测移植后14、28d缺血侧脑组织BDNFmRNA表达水平。结果(1)移植后3d2组Bederson评分无显著差别,移植后14、28d治疗组显著低于对照组(P<0.05)。(2)移植后3d两组脑梗死面积无显著差别,移植后14、28d治疗组显著低于对照组(P<0.05)。(3)治疗组移植后3d梗死侧见少量DAPi阳性染色细胞,移植后14、28d梗死侧DAPi阳性染色细胞增加(P<0.05)...     

骨髓间质干细胞移植减轻犬急性脑梗死模型的缺血性脑损伤研究

目的观察骨髓间质干细胞(BMSCs)对犬急性缺血脑组织的保护作用,并探讨其可能的机制。方法将24只成年杂交犬随机分为治疗组及对照组,DSA引导下行自体血栓栓塞大脑中动脉闭塞缺血模型制作,并抽取骨髓提取BMSCs,传代并给予4’,6-二脒基-2-苯基吲哚(DAPI)标记;1周后开颅行多点脑内注射BMSCs移植;移植后1周行脑DWI序列扫描,计算梗死灶体积;4周后将犬处死,每组随机选择6只动物取脑标本行TTC染色测定梗死灶体积;另外6只动物进行HE染色、VG染色、TUNEL染色评价脑损伤情况;免疫荧光染色了解脑源性神经营养因子(BDNF)、碱性成纤维生长因子(b FGF)、胰岛素样生长因子1(IGF-1)和血管内皮生长因子(VEGF)的表达情况。结果治疗组梗死灶内广泛存在DAPI阳性细胞。治疗组的梗死灶体积明显小于对照组,P0.01。治疗组梗死灶范围、梗死灶内细胞坏死、胶质增生和胶质瘢痕均较对照组减轻。治疗组凋亡细胞明显少于对照组,P0.05。治疗组细胞因子BDNF、BFGF、IGF-1和VEGF表达阳性的细胞均显著多于对照组。结论脑梗死后给予BMSCs移植,BMSCs能存活并自行向梗死灶迁移,并发挥脑保护作用,其机制可能和分泌多种神经营养因子有关。     

生物波调控因子对实验性脑梗死大鼠脑组织IL-6表达的影响

目的:探讨生物波调控因子(BRF)对实验性脑梗死大鼠脑组织IL-6表达的影响。方法:成年健康雄性SD大鼠90只随机分为BRF组、生理盐水组和假手术组。制备大脑中动脉梗死(MCAO)模型,术后1h以1ml/100g的剂量分别腹腔注射1.25%BRF溶液和生理盐水,此后1次/d。进行行为学评分、测定脑组织含水量、HE染色观察组织病理学改变、免疫组织化学方法测定脑组织IL-6的动态变化。结果:生理盐水组及BRF治疗组脑梗死组织周围组织含水量于24h开始明显升高,48h达高峰,持续至72h,7d时明显下降,在各个时间点与假手术组比较均有统计学意义。除假手术组外,其余各组梗死区炎细胞浸润和IL-6阳性细胞表达于梗死后6h开始增多,48h达高峰,并持续至7d;假手术组未见明显炎细胞浸润,可见少量IL-6阳性细胞。BRF治疗组大鼠行为学评分降低,脑组织水肿程度减轻,术后48h最显著,病理损伤减轻,脑组织IL-6阳性细胞减少,术后48h、72h最显著。结论:生物波调控因子可以通过减轻梗死后脑水肿,降低脑内IL-6的表达,从而对大鼠缺血性脑组织损伤产生保护作用。     

丹参多酚酸盐对缺血再灌注损伤大鼠脑组织BDNF和GDNF的影响

目的观察丹参多酚酸盐对大鼠局灶性脑缺血再灌注损伤后脑组织内脑源性神经营养因子（BDNF）及胶质源性神经营养因子（GDNF）表达的影响,探讨其神经保护作用的机制。方法线栓法制作大鼠大脑中动脉闭塞再灌注损伤模型,SD大鼠随机分为正常对照组、缺血再灌注组、丹参多酚酸盐低剂量治疗组（10mg／kg）和丹参多酚酸盐高剂量治疗组（30mg／kg）,观察各组大鼠的行为学改变,测定脑梗死体积,ELISA法检测脑组织中BDNF和GDNF的含量。结果与缺血再灌组比较,丹参多酚酸盐可以显著降低大鼠神经损伤行为学评分,减少脑梗死体积,增加脑组织内BDNF和GDNF的含量,且呈剂量依赖性。结论丹参多酚酸盐对脑缺血再灌注损伤有明显的保护作用,其机制与促进脑组织合成BDNF及GDNF有关。     

缺血再灌流时胶质源性神经营养因子在大鼠脑组织的分布特点

目的 观察大鼠脑缺血再灌流时胶质源性神经营养因子（GDNF）在脑组织的分布特点，及其在缺血性脑损伤中的作用。方法 阻断大鼠大脑中动脉（MCA）血流2小时，再灌流0．5－48小时制成局灶性脑缺血模型，HE染色评价缺血性脑损伤的组织学特点，免疫组化法观察GDNF在脑组织的分布特点。结果 再灌流0．5小时组有灶性缺血区，24小时组面积最大，包括视前区、纹状体和皮质。再灌注6小时组开始出现神经元不可逆变性，24小时组梗死形成。再灌注0．5小时组，缺血区皮质神经元GDNF弱阳性，缺血周边区中等阳性；再灌流3－48小时组，缺血区神经元GDNF阴性。再灌流48小时组视前区的梗死周边区巨噬细胞GDNF呈强阳性。GDNF阳性细胞计数显示缺血区各实验组与正常组相比均减少（均P＜0．01）；24小时和48小时组分别与0．5小时组和3小时组相比，GDNF阳性细胞数减少（分别P＜0．01）。结论 缺血性脑损伤时，变性死亡的神经元GDNF不表达，存活的神经元和活化的小胶质细胞或巨噬细胞的GDNF表达增加。     

侧脑室注射质粒pLXSN介导bcl-2基因对大鼠脑梗死及BDNF影响的研究

目的 研究大鼠短暂大脑中动脉闭塞（MCAO）模型经侧脑室注射质粒pLXSN介导bcl-2基因后，该基因对脑梗死及脑源性神经营养因子（brain-derived neurotrophic factor,BDNF）表达的影响。方法取135只Wistar大鼠随机分配为3组,每组45只．对照组为脑室注射生理盐水和空质粒两组．治疗组为脑室注射Bcl-质粒pLXSN介导bcl-2基因。以改良线栓法制备短暂缺血2h脑梗死模型。每组分别于缺血后3、6、24、48、72h为时间点。分别测量梗死体积变化以及BDNF蛋白的表达。结果MCAO后24、48、72h．梗死体积治疗组显著小于对照组（P〈0．05）；3、6h时间点无显著变化（P〉0．05）。MCAO后24、48、72h。治疗组Bcl-2和BDNF蛋白的表达较对照组显著增加（P〈0．01）．而3、6h时间点无显著改变（P〉0．05）。结论脑室注射质粒pLXSN介导bcl-2基因对短暂脑缺血有治疗作用．并可增加BDNF表达。上调BDNF表达所起的神经保护作用可能是短暂脑缺血后bcl-2基因的治疗作用机制之一。     

骨髓基质细胞及血管内皮祖细胞共移植对局灶性脑缺血大鼠系列生长因子表达的影响

目的 观察经静脉移植骨髓基质细胞(BMSCs)及血管内皮祖细胞(EPCs)后血管内皮生长因子(VEGF)、基质细胞衍生因子-1(SDF-1)、碱性成纤维生长因子(bFGF)、胰岛素样生长因子-1(IGF-1)、转化生长因子β(TGF-β)、血小板源性生长因子(PDGF)、脑源性生长因子(BDNF)、胶质源性生长因子(GDNF)和神经生长因子(NGF)这9种细胞因子的表达. 方法 80只健康成年Wistar 大鼠按照随机数字表法分为模型对照组、BMSCs移植组、EPCs移植组及联合移植组,每组20只.线栓法建立大鼠大脑中动脉闭塞模型.造模24h后,分别取浓度为3x106个/mL的BMSCs、EPCs悬液及两者混合液1mL经鼠尾静脉移植入BMSCs移植组、EPCs移植组及联合移植组;模型对照组鼠尾静脉注射1 mL生理盐水.移植后7d,实时荧光PCR、Western boltting检测各组大鼠脑内生长因子的表达. 结果 联合移植组中bFGF、VEGF、BDNF mRNA表达水平最高,与其他3组比较差异均有统计学意义(P＜0.05).BMSCs移植组中NGF、GDNF、TGF-β mRNA表达水平最高,与其他3组比较差异均有统计学意义(P＜0.05),联合移植组次之.EPCs组中PDGF-BB、IGF-1、SDF-1 mRNA表达水平最高,与其他3组比较差异均有统计学意义(P＜0.05),联合移植组次之. 结论 BMSCs及EPCs联合静脉移植可上调脑内生长因子表达水平,为提高细胞移植效率提供了新的尝试途径.     

Differential expression of GDNF, BDNF, and NT-3 in the aging nigrostriatal system following a neurotoxic lesion

Protein levels for brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and glial cell line-derived neurotrophic factor (GDNF) were measured in the striatum and ventral midbrain of young and aged Brown Norway/F344 F1 (F344BNF(1)) hybrid rats following a unilateral 6-hydroxydopamine (6-OHDA) lesion of the nigrostriatal pathway. At 2 weeks post-lesion, protein levels of BDNF and GDNF were higher in the denervated striatum when compared to the intact striatum for young (4-5 months old) but not old (31-33 months old) rats. Interestingly, in old rats BDNF protein in the denervated striatum was significantly lower than that measured in the intact striatum. At the same time point BDNF protein levels in the ventral midbrain were higher on the lesioned versus intact side for both young and old rats while no significant side differences were detected for GDNF protein in the ventral midbrain of young or old rats. No significant differences in NT-3 protein levels were detected between the lesioned and intact sides for striatal or ventral midbrain regions in either young or old brain. While no significant age effects were detected for BDNF or NT-3 protein, young rats showed higher GDNF protein levels in both the striatum (lesioned or intact) and ventral midbrain (lesioned or intact) than old rats. These data show that two endogenous neurotrophic factors, BDNF and GDNF, are differentially affected by a 6-OHDA lesion in the aging nigrostriatal system with young brain showing a significant compensatory increase of these two factors in the denervated striatum while no compensatory increase is observed in aged brain.     

Human neural progenitor cells over-expressing IGF-1 protect dopamine neurons and restore function in a rat model of Parkinson's disease

Growth factors such as glial cell line-derived neurotrophic factor (GDNF) have been shown to prevent neurodegeneration and promote regeneration in many animal models of Parkinson's disease (PD). Insulin-like growth factor 1 (IGF-1) is also known to have neuroprotective effects in a number of disease models but has not been extensively studied in models of PD. We produced human neural progenitor cells (hNPC) releasing either GDNF or IGF-1 and transplanted them into a rat model of PD. hNPC secreting either GDNF or IGF-1 were shown to significantly reduce amphetamine-induced rotational asymmetry and dopamine neuron loss when transplanted 7 days after a 6-hydroxydopamine (6-OHDA) lesion. Neither untransduced hNPC nor a sham transplant had this effect suggesting GDNF and IGF-1 release was required. Interestingly, GDNF, but not IGF-1, was able to protect or regenerate tyrosine hydroxylase-positive fibers in the striatum. In contrast, IGF-1, but not GDNF, significantly increased the overall survival of hNPC both in vitro and following transplantation. This suggests a dual role of IGF-1 to both increase hNPC survival after transplantation and exert trophic effects on degenerating dopamine neurons in this rat model of PD.     

Inhibition of brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor expression reduces dopaminergic sprouting in the injured striatum

After striatal injury, sprouting dopaminergic fibres grow towards and intimately surround wound macrophages which, together with microglia, express the dopaminergic neurotrophic factors glial cell line-derived neurotrophic factor (GDNF) and brain derived neurotrophic factor (BDNF). To evaluate the importance of these endogenously secreted neurotrophic factors in generating striatal peri-wound dopaminergic sprouting, the peri-wound expression of BDNF or GDNF was inhibited by intrastriatal infusion of antisense oligonucleotides for 2 weeks in mice. Knock-down of both BDNF and GDNF mRNA and protein levels in the wounded striatum were confirmed by in situ hybridization and enzyme-linked immunosorbent assay, respectively. Dopamine transporter immunohisto-chemistry revealed that inhibition of either BDNF or GDNF expression resulted in a marked decrease in the intensity of peri-wound sprouting. Quantification of this effect using [H3]-mazindol autoradiography confirmed that peri-wound sprouting was significantly reduced in mice receiving BDNF or GDNF antisense infusions whilst control infusions of buffered saline or sense oligonucleotides resulted in the pronounced peri-wound sprouting response normally associated with striatal injury. BDNF and GDNF thus appear to be important neurotrophic factors inducing dopaminergic sprouting after striatal injury.     

Granulocyte colony‐stimulating factor promotes behavioral recovery in a mouse model of traumatic brain injury

Hematopoietic growth factors such as granulocyte colony‐stimulating factor (G‐CSF) represent a novel approach for treatment of traumatic brain injury (TBI). After mild controlled cortical impact (CCI), mice were treated with G‐CSF (100 μg/kg) for 3 consecutive days. The primary behavioral endpoint was performance on the radial arm water maze (RAWM), assessed 7 and 14 days after CCI. Secondary endpoints included 1) motor performance on a rotating cylinder (rotarod), 2) measurement of microglial and astroglial response, 3) hippocampal neurogenesis, and 4) measures of neurotrophic factors (brain‐derived neurotrophic factor [BDNF] and glial cell line‐derived neurotrophic factor [GDNF]) and cytokines in brain homogenates. G‐CSF‐treated animals performed significantly better than vehicle‐treated mice in the RAWM at 1 and 2 weeks but not on the rotarod. Cellular changes found in the G‐CSF group included increased hippocampal neurogenesis as well as astrocytosis and microgliosis in both the striatum and the hippocampus. Neurotrophic factors GDNF and BDNF, elaborated by activated microglia and astrocytes, were increased in G‐CSF‐treated mice. These factors along with G‐CSF itself are known to promote hippocampal neurogenesis and inhibit apoptosis and likely contributed to improvement in the hippocampal‐dependent learning task. Six cytokines that were modulated by G‐CSF treatment following CCI were elevated on day 3, but only one of them remained altered by day 7, and all of them were no different from vehicle controls by day 14. The pro‐ and anti‐inflammatory cytokines modulated by G‐CSF administration interact in a complex and incompletely understood network involving both damage and recovery processes, underscoring the dual role of inflammation after TBI. © 2016 Wiley Periodicals, Inc.     

微囊化猪视网膜色素上皮细胞移植治疗帕金森病的实验研究(英文)

目的研究微囊化后的猪视网膜色素上皮细胞(retinal pigment epithelial,RPE)对帕金森病大鼠模型的移植疗效。方法原代培养RPE 并传代,高效液相色谱法测定培养液上清中多巴胺(dopamine, DA)和高香草酸(homovanillic acid, HVA)的含量,ELISA法检测脑源性神经营养因子(brain-derived neurotrophic factor, BDNF)和胶质细胞源性神经营养因子(glial-derived neurotrophic factor, GDNF)的含量。用高压静电成囊装置制备海藻酸钠-多聚赖氨酸-海藻酸钠微囊化细胞。6-羟基多巴胺(6-hydroxydopamine, 6-OHDA)毁损内侧前脑束 (medial fore-brain bundle,MFB)建立 SD 大鼠帕金森病模型。立体定向移植 RPE+ 微囊,检验旋转实验、免疫组化和脑内生化的变化。结果 RPE 培养上清液中DA、HVA、BDNF、GDNF 的含量稳定,微囊化后细胞长期存活,活性没有明显变化。6-OHDA毁损MFB建立大鼠帕金森病模型的成模率为83%。移植微囊化的RPE后有效率为33%。结论猪 RPE 体外培养生长旺盛,持续分泌 DA、BDNF 和 GNDF,微囊化不影响其分泌功能。RPE 移植对帕金森病大鼠有一定的治疗作用。     

Effects of GDNF-Transfected Marrow Stromal Cells on Rats with Intracerebral Hemorrhage

Objective: The present study aimed to investigate the effects of Mesenchymal stem cells/glial cell line derived neurotrophic factor (MSCs/GDNF) transplantation on nerve reconstruction in rats with intracerebral hemorrhage. Methods: GDNF transduction to MSCs was using adenovirus vector pAdEasy-1-pAdTrack-CMV prepared. Intracerebral hemorrhage (ICH) was induced by injection of collagenase and heparin into the caudate putamen. At the third day after a collagenase-induced ICH, adult male SD rats were randomly divided into saline group, MSCs group and MSCs/GDNF group. Immunofluorescence and RT-PCR were performed to detect the differentiation of MSCs or MSCs with an adenovirus vector encoding GDNF gene in vivo and in vitro. Result: After 6 hours of induction, both MSCs and MSCs/GDNF expressed neuro or glial specific markers and synaptic-associated proteins (SYN, GAP-43, PSD-95); additionally, they secreted bioactive compounds (BDNF, NGF-β). MSCs/GDNF transplantation, compared to MSCs and saline solution injection, significantly improved neurological functions after ICH. The grafted MSCs or MSCs/GDNF survived in the striatum after 2 weeks of transplantation and expressed the neural cell-specific biomarkers NSE, MAP2, and GFAP. Conclusion: These findings demonstrate that MSCs/GDNF transplantation contributes to improved neurological function in experimental ICH rats. The mechanisms are possibly due to neuronal replacement and enhanced neurotrophic factor secretion.     

Changes in spinal GDNF, BDNF, and NT-3 expression after transient spinal cord ischemia in the rat

Previous studies have demonstrated that the expression of several growth factors including glial cell-derived neurotrophic factor (GDNF), brain-derived growth factor (BDNF), and neurotrophin-3 (NT-3) play an important role in defining neuronal survival after brain ischemia. In the present study, using a well-defined model of transient spinal ischemia in rat, we characterized the changes in spinal GDNF, BDNF, and NT-3 expression as defined by enzyme-linked immunosorbent assay (ELISA) and immunofluorescence coupled with deconvolution microscopy. In control animals, baseline levels of GDNF, BDNF, and NT-3 (74 +/- 22, 3,600 +/- 270, 593 +/- 176 pg/g tissue, respectively) were measured. In the ischemic group, 6 min of spinal ischemia resulted in a biphasic response with increases in tissue GDNF and BDNF concentrations at the 2-hr and 72-hr points after ischemia. No significant differences in NT-3 concentration were detected. Deconvolution analysis revealed that the initial increase in tissue GDNF concentration corresponded to a neuronal upregulation whereas the late peak seen at 72 hr corresponded with increased astrocyte-derived GDNF synthesis. Increased expression of BDNF was seen in neurons, astrocytes, and oligodendrocytes. These data suggest that the early increase in neuronal GDNF/BDNF expression likely modulates neuronal resistance/recovery during the initial period of postischemic reflow. Increased astrocyte-derived BDNF/GDNF expression corresponds with transient activation of astrocytes and may play an active role in neuronal plasticity after non-injurious intervals of spinal ischemia.     

微囊化猪视网膜色素上皮细胞移植治疗帕金森病的实验研究

目的研究微囊化后的猪视网膜色素上皮细胞（retinal pigment epithelial,RPE）对帕金森病大鼠模型的移植疗效。方法原代培养RPE并传代,高效液相色谱法测定培养液上清中多巴胺（dopamine,DA）和高香草酸（homovanillic acid,HVA）的含量,ELISA法检测脑源性神经营养因子（brain-derived neurotrophic factor,BDNF）和胶质细胞源性神经营养因子（glial-derived neurotrophic factor,GDNF）的含量。用高压静电成囊装置制备海藻酸钠-多聚赖氨酸-海藻酸钠微囊化细胞。6-羟基多巴胺（6-hydroxydopamine,6-OHDA）毁损内侧前脑束（medial forebrain bundle,MFB）建立SD大鼠帕金森病模型。立体定向移植RPE＋微囊,检验旋转实验、免疫组化和脑内生化的变化。结果RPE培养上清液中DA、HVA、BDNF、GDNF的含量稳定,微囊化后细胞长期存活,活性没有明显变化。6-OHDA毁损MFB建立大鼠帕金森病模型的成模率为83％。移植微囊化的RPE后有效率为33％。结论猪RPE体外培养生长旺盛,持续分泌DA、BDNF和GNDF,微囊化不影响其分泌功能。RPE移植对帕金森病大鼠有一定的治疗作用。     

Glial cell line‐derived neurotrophic factor–secreting genetically modified human bone marrow‐derived mesenchymal stem cells promote recovery in a rat model of Parkinson's disease

Parkinson's disease (PD) is a neurodegenerative disease characterized by progressive degeneration of nigrostriatal dopaminergic (DA) neurons. The therapeutic potential of glial cell line‐derived neurotrophic factor (GDNF), the most potent neurotrophic factor for DA neurons, has been demonstrated in many experimental models of PD. However, chronic delivery of GDNF to DA neurons in the brain remains an unmet challenge. Here, we report the effects of GDNF‐releasing Notch‐induced human bone marrow‐derived mesenchymal stem cells (MSC) grafted into striatum of the 6‐hydroxydopamine (6‐OHDA) progressively lesioned rat model of PD. Human MSC, obtained from bone marrow aspirates of young, healthy adult volunteers, were transiently transfected with the intracellular domain of the Notch1 gene (NICD) to generate SB623 cells. SB623 cells expressing GDNF and/or humanized Renilla green fluorescent protein (hrGFP) following lentiviral transduction or nontransduced cells were stereotaxically placed into rat striatum 1 week after a unilateral partial 6‐OHDA striatal lesion. At 4 weeks, rats that had received GDNF‐transduced SB623 cells had significantly decreased amphetamine‐induced rotation compared with control rats, although this effect was not observed in rats that received GFP‐transduced or nontransduced SB623 cells. At 5 weeks, rejuvenated tyrosine hydroxylase‐immunoreactive (TH‐IR) fibers that appeared to be host DA axons were observed in and around grafts. This effect was more prominent in rats that received GDNF‐secreting cells and was not observed in controls. These observations suggest that human bone‐marrow derived MSC, genetically modified to secrete GDNF, hold potential as an allogeneic or autologous stem cell therapy for PD. © 2010 Wiley‐Liss, Inc.