鼠神经生长因子对急性脑梗死患者外周血EPCs、VEGF及SDF-1水平的影响

目的 探讨鼠神经生长因子(mNGF)对急性脑梗死患者外周血EPCs、VEGF及SDF-1水平的影响。方法 将130例急性脑梗死患者随机分为对照组和观察组,每组各65例; 对照组给予活血祛瘀、抑制血小板聚集、改善微循环等常规治疗,观察组在常规治疗基础上加用鼠NGF肌肉注射治疗,30 μg/次,1次/d; 2组均治疗21 d,比较2组治疗后总有效率、梗死面积、美国国立卫生研究院脑卒中量表(NIHSS)评分、格拉斯哥昏迷指数(GCS)评分及外周血内皮祖细胞(EPCs)、血管内皮生长因子(VEGF)、基质细胞衍生因子-1(SDF-1)水平。结果 ①观察组治疗总有效率为83.08%,明显高于对照组的67.69%(P<0.05); ②与治疗前比较,2组治疗后梗死面积、NIHSS评分明显降低(P<0.05),GCS 评分明显增高(P<0.05),且观察组变化幅度显著大于对照组(P<0.05); ③2组治疗前外周血EPCs数量、VEGF及SDF-1水平无明显差异(P>0.05),观察组治疗后EPCs数量、VEGF及SDF-1水平显著高于对照组同期水平(P<0.05)。结论 mNGF 可促进急性脑梗死患者外周血EPCs、VEGF及SDF-1水平表达,从而有效改善脑梗死后神经功能缺损症状。     

缺氧诱导因子-1α对脑梗死后内源性神经干细胞作用的研究进展

神经干细胞(neural stem cells,NSCs)治疗脑梗死无时间窗限制,无出血风险,且移植的细胞可以分泌神经生长因子改善微环境,抑制炎症反应,减少细胞凋亡,促进神经再生,重建受损的神经网络。缺氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)是脑缺血缺氧损伤中起神经保护作用的关键因子,其在脑梗死后促进NSCs再生的作用广泛而复杂。本文将系统综述HIF-1α在脑缺血缺氧损伤后对促进内源性NSCs增值、迁移、分化方面所起的作用,说明其可能成为临床治疗脑梗死的有效作用靶点。     

基质细胞衍生因子-1诱导神经干细胞靶向性迁移的实验研究

目的 观察基质细胞衍生因子-1(SDF-1)对神经干细胞(NSCs)迁移的影响.方法 由胚胎大鼠脑组织获取NSCs并进行传代培养和分化鉴定,使用流式细胞术检测NSCs纯度及SDF-1特异性受体CXCR4的表达情况,之后利用Blind-Well小室体外迁移体系观察不同浓度的SDF-1(0 ng/L、1 ng/mL、10 ng/mL、50 ng/mL、100 ng/mL、500 ng/mL和1000 ng/mL)对NSCs迁移数量的影响,并使用μ-载片观察SDF-1对NSCs迁移距离的影响.结果成功分离培养得到了能够在体外不断分裂增殖、具有多向分化潜能的NSCs,连续传3代后绝大部分细胞nestin表达阳性,nestin和CXCR4的共表达率达到80%左右.细胞趋化实验结果表明,SDF-1对NSCs有较强的趋化作用,随着SDF-1浓度的升高,发生迁移的细胞数量也随之增加,并于SDF-1浓度为500ng/mL时达到最高峰[(256.79±38.27)个细胞/每高倍视野].在μ-载片细胞生长通道两侧的SDF-1浓度梯度(500ng/mL→0ng/mL)作用下,由细胞克隆球迁出的细胞呈不对称分布,通常有更多的迁出细胞分布于趋化因子浓度较高的一侧,且该侧细胞的最大迁移距离也比对侧远.结论 SDF-1与其特异性受体CXCR4相巨作用能够诱导NSCs发生靶向性迁移.     

基质细胞衍生因子-1诱导神经干细胞靶向性迁移的实验研究

Objective To investigate the effect ofstromal cell derived factor-1 (SDF-1) on the regulation of neural stem cells (NSCs) migration.Methods NSCs were obtained from the cerebral cortex of embryonic rats and cultured in serum-free medium,and their stem cell properties were assessed by means of induced differentiation in vitro into neurons and astrocytes.After in vitro cell culture,the purity of NSCs and the co-expression rate of CXCR4/nestin were detected by flow cytometry.Blind-well chambers were employed to detect the chemotactic effects of SDF-1 by counting the cells which had crossed a 8 μm pore membrane when confronted with varying concentrations of SDF-1 (0,1,10,50,100,500 and 1000 ng/mL),and the distribution of cells migrated out of the same neurosphere was overviewed by μ-slides in the persistent concentration gradient of SDF-1.Results Neurospheres were formed by persistent proliferation of NSCs, which were capable of differentiating into neurons (β-tubulin+) and astrocytes (GFAP+) in media without mitogens,and flow cytometry analyses showed that most of the cultured cells expressed nestin and the co-expression rate of CXCR4/nestin was nearly 80%.SDF-1 showed great chemotaxis to NSCs,and the amount of cells having migrated through the membrane in 500 ng/ml SDF-1 group was higher than that in other groups (P<0.05).When the cells were confronted with a linear concentration gradient (from 500 to 0 ng/mL),which was generated by diffusion and stable for at least 48 h,the cells migrated out ofa neruosphere could distribute irregularly with more cells locating in the region of higher concentration of SDF-1 and longer migration distance away from the center of the neurosphere than the opposite.Conclusion SDF-1 binding to its specific receptor CXCR4 was capable of inducing NSCs migrating directionally to the source of SDF-1.     

基质细胞衍生因子-1诱导神经干细胞靶向性迁移的实验研究

Objective To investigate the effect ofstromal cell derived factor-1 (SDF-1) on the regulation of neural stem cells (NSCs) migration.Methods NSCs were obtained from the cerebral cortex of embryonic rats and cultured in serum-free medium,and their stem cell properties were assessed by means of induced differentiation in vitro into neurons and astrocytes.After in vitro cell culture,the purity of NSCs and the co-expression rate of CXCR4/nestin were detected by flow cytometry.Blind-well chambers were employed to detect the chemotactic effects of SDF-1 by counting the cells which had crossed a 8 μm pore membrane when confronted with varying concentrations of SDF-1 (0,1,10,50,100,500 and 1000 ng/mL),and the distribution of cells migrated out of the same neurosphere was overviewed by μ-slides in the persistent concentration gradient of SDF-1.Results Neurospheres were formed by persistent proliferation of NSCs, which were capable of differentiating into neurons (β-tubulin+) and astrocytes (GFAP+) in media without mitogens,and flow cytometry analyses showed that most of the cultured cells expressed nestin and the co-expression rate of CXCR4/nestin was nearly 80%.SDF-1 showed great chemotaxis to NSCs,and the amount of cells having migrated through the membrane in 500 ng/ml SDF-1 group was higher than that in other groups (P<0.05).When the cells were confronted with a linear concentration gradient (from 500 to 0 ng/mL),which was generated by diffusion and stable for at least 48 h,the cells migrated out ofa neruosphere could distribute irregularly with more cells locating in the region of higher concentration of SDF-1 and longer migration distance away from the center of the neurosphere than the opposite.Conclusion SDF-1 binding to its specific receptor CXCR4 was capable of inducing NSCs migrating directionally to the source of SDF-1.     

EGF对神经干细胞迁移的影响

目的研究表皮生长因子(EGF)对神经干细胞迁移的影响。方法取孕13—14dSD大鼠胚胎纹状体行神经干细胞培养，7d后取悬浮神经球制成单细胞悬液再培养，EGF作用下连续培养14d后分为实验组和对照组，继续培养，观察：EGF对神经干细胞迁移的影响。结果原代培养的细胞球是神经干细胞，EGF传代培养14d时细胞球大小约100个细胞左右，实验组观察到第14d细胞球增殖变慢，细胞球长出突起与邻近的细胞球相接触，14—17d观察到神经干细胞迁移，17d后细胞迁移现象消失；对照组中未观察到细胞迁移。结论：EGF在某一特定时间内能够诱导神经干细胞迁移。     

基质细胞衍生因子1趋化神经干细胞迁移的体外效应

背景：神经干细胞的迁移在神经系统的发育和损伤修复中起着至关重要的作用,近来研究表明趋化因子参与神经干细胞的迁移,但关于其迁移机制目前尚不清楚。 目的：观察体外条件下基质细胞衍生因子1对胎鼠海马神经干细胞的趋化迁移作用。 方法：通过无血清法体外分离、培养及鉴定胎鼠海马神经干细胞;细胞免疫荧光及RT-PCR检测其CXCR4是否表达;观察不同浓度基质细胞衍生因子1对神经干细胞的趋化迁移作用,中和CXCR4受体以验证基质细胞衍生因子1趋化迁移作用的特异性。 结果与结论：胎鼠海马来源神经干细胞表达趋化因子受体CXCR4,呈阳性。RT-PCR琼脂糖凝胶电泳后出现643 bp特异性扩增条带。体外条件下基质细胞衍生因子1趋化迁移随浓度而增强,500 μg/L为最佳趋化浓度。加入抗CXCR4多克隆抗体中和后,神经干细胞迁移较基质细胞衍生因子1组明显减少,与对照组比较,差异无显著性意义(P > 0.05),提示抗CXCR4多克隆抗体可阻断趋化迁移作用。     

血清SDF-1、Ang-1与急性缺血性脑卒中预后的关系及其预测价值

目的 探讨急性缺血性脑卒中(AIS)患者血清基质细胞衍生因子-1(SDF-1)、血管生成素-1(Ang-1)水平变化的临床意义。方法 选取2017年2月-2019年2月本院收治的AIS患者107例作为观察组,并根据mRS评分将观察组分为预后良好组(n=72, mRS评分≤2分)和预后不良组(n=35, mRS评分>2分); 另选取同期来本院行体检的健康志愿者50例为对照组; 比较对照组、观察组不同预后的血清SDF-1、Ang-1水平的动态变化; 采用多元 Logistic回归分析预后的影响因素; 采用ROC曲线评估SDF-1、Ang-1水平对患者预后的预测价值。结果 与对照组比较,观察组入院第1、7、14、21 d的SDF-1、Ang-1水平均存在一定的波动性变化(P<0.05)。预后良好组入院第1、7、14、21 d的SDF-1水平低于预后不良组,Ang-1水平高于预后不良组(P<0.05)。单因素分析显示,房颤、大面积脑梗死、血尿酸、血肌酐、血浆脑钠肽水平、NIHSS评分与AIS预后有关(P<0.05)。多因素Logistic回归分析显示,血清SDF-1水平升高、Ang-1水平降低、大面积脑梗死、房颤、血浆脑钠肽水平≥280 ng/L、NIHSS评分≥13分是AIS预后的独立危险因素(P<0.05)。ROC分析显示,SDF-1、Ang-1水平用于预测AIS患者预后的AUC、敏感度、特异度均高于0.7,二者联合检测的敏感度和特异度分别为0.857(30/35),0.819(59/72),均高于两指标单独检测。结论 AIS患者血清SDF-1、Ang-1水平呈异常波动,且其表达水平与患者的预后密切相关,二者联合检测对患者预后有较高的预测价值     

VEGF在人胚胎干细胞向神经元分化中作用的研究

目的观察血管内皮生长因子（vascular endothelial growth factor,VEGF）在人胚胎干细胞分化为神经元过程中是否发挥作用及相关机制。方法人胚胎干细胞经拟胚体向神经元分化,分为3组：A组为常规诱导组,B组为常规诱导＋VEGF（10ng／mL）作用组,C组为常规诱导＋VEGF（10ng／mL）＋VEGFR2／Fc嵌合体（10ng／mL）作用组;用RT-PCR、免疫荧光法检测各阶段细胞标志物,计算并用流式细胞仪检测各组不同阶段细胞阳性率。结果用RT-PCR分别检测到OCT4、Nestin、MAP2表达;免疫荧光法检测显示B组产生神经干细胞、分化为神经元的阳性率明显高于A、C两组,差异有统计学意义（P〈0．01）,A、C两组之间无显著性差异（P〉0．05）;流式细胞仪检测与免疫荧光法相似。结论人胚胎干细胞体外分化过程中血管内皮生长因子通过血管内皮生长因子受体2来促进神经干细胞增殖及向神经元分化。     

人胚胎干细胞神经分化中VEGF作用的分子机制研究

目的 观察血管内皮生长因子(vascular endothelial growth factor,VEGF)对人胚胎干细胞分化为神经元的促进作用是否与p38MAPK信号通路相关.方法 人胚胎干细胞经拟胚体向神经元分化,分为3组:A组:常规诱导组;B组:常规诱导+VEGF(10 ng/mL)作用组;C组:常规诱导+p38...     

趋化因子SDF-1体外趋化骨髓基质细胞迁移的实验研究

目的探讨趋化因子SDF-1在体外对骨髓基质细胞的迁移作用。方法采用全骨髓法培养成年Wistar大鼠骨髓基质细胞。取第五代骨髓基质细胞行免疫荧光鉴定；后通过细胞免疫荧光及RT-PCR的方法检测骨髓基质细胞表达趋化因子受体CXCR4的情况．利用Boyden小室法探讨趋化因子SDF-1对骨髓基质细胞的体外趋化作用及其特异性。结果第五代骨髓基质细胞都表达间充质干细胞标记物Vimentin、Laminin及Fibronectin；细胞免疫荧光及RT-PCR结果证实骨髓基质细胞表达趋化因子受体CXCR4；趋化因子SDF-1（5、50、500ng／mL）体外可趋化骨髓基质细胞迁移，抗SDF-1多克隆抗体可对抗其趋化迁移作用。结论SDF-1／CXCR4通路参与骨髓基质细胞体外迁移，为进一步研究骨髓基质细胞的迁移机制提供了理论依据。     

Effect of epidermal growth factor on the migration of neural stem cells

INTRODUCTION Since neural stem cells (NSCs) are separated from corpus striatum of adult rats in 1992 [1], researches on NSCs become a hot topic all over the world because they are characterized by excellent superiority and complete cure on treating geneti…     

神经干细胞延伸于脑肿瘤干细胞的研究

1992年Reynolds首先成功地从成年小鼠纹状体中分离出神经干细胞（neural stem cells，NSCs）；1997年Mckay确立了神经干细胞能提供大量脑组织所需要的功能细胞的概念；2001年Reya提出了肿瘤干细胞理论：Singh等在人脑胶质瘤组织中分离出神经干细胞样的肿瘤干细胞，并进一步证明后者可直接生成肿瘤细胞，目前称之为脑肿瘤干细胞（brain tumor stem cells，BTSCs），由此开创了神经干细胞研究的新领域。[第一段]     

纤维连接蛋白促进人神经球内干细胞的迁移

目的研究细胞外基质成份纤维连接蛋白(Fn)对人神经干细胞迁移的作用。方法用Fn包被培养孔,于细胞球转种后的24h、48h、72h、96h、120h、144h、168h和240h各时间点动态观察神经球细胞的迁移变化,测量细胞迁移的最远距离,计算迁移速率,用荧光免疫技术检测迁移细胞的特异性表型标志,计算一定视野内nestin+细胞所占的百分比,同时,平行观察含胎牛血清(FBS)培养诱导的迁移。设立多聚赖氨酸(Ply)包被组以及空白组作为实验对照。结果Ply包被组以及空白组均无细胞迁移发生,FBS组和Fn组的细胞球有细胞从球体向外迁移,二者迁移速率均在前48h达最高峰,但迁移方式不同。Fn组迁移的细胞分化不明显,细胞分裂相多见,nestin+细胞占(30.60±8.06)%,而FBS组迁移出来的细胞成进行性分化梯度,偶见有nestin+细胞,外周边缘区域GFAP+星形胶质细胞是其主要细胞群。结论Fn在体外可促进人胎脑神经干细胞的迁移。     

Immunomodulation by neural stem cells

Neural (stem) cell transplantation has been proposed as a means of cell replacement therapy. Multipotential neural precursor cells (NPCs) that expand in floating spheres, and are (partially) committed to a glial fate, showed excellent remyelinating properties in a focal, chemically induced demyelinated lesion in the rat spinal cord. When transplanted into the CNS of rodents with acute and chronic EAE the NPCs were attracted by the inflammatory process to migrate exclusively into inflamed white matter but not into adjacent gray matter. Following magnetic labeling, mouse NPCs and human ESC-derived neural precursors' migration was detected by high-resolution magnetic resonance images. Intraventricular transplantation of neural spheres attenuated brain inflammation in acute and chronic EAE, reduced the clinical severity of disease, and reduced demyelination and axonal pathology. Intravenous (IV) NPC injection also inhibited EAE and reduced CNS inflammation and tissue injury. However, NPCs did not enter the CNS but were transiently found in lymph nodes and spleen, where they inhibited the activation and proliferation of T cells and markedly reduced their encephalitogenicity. Thus, IV administration of neural precursors inhibits EAE by a peripheral immunosuppression, involving a profound bystander inhibitory effect of NPCs on T cell activation and proliferation in lymph nodes. In conclusion, neural precursor cells exert an immunomodulatory effect that inhibits CNS inflammation. Cell therapy in MS should be optimized to utilize both regenerative and immunologic properties of the cells.     

三种底物对体外培养的神经干细胞分化和迁移能力的影响

目的:探讨不同底物对体外培养的神经干细胞分化和迁移能力的影响.方法:采用多聚鸟氨酸、层黏连蛋白、鼠尾胶原等作为底物,观察它们对大鼠神经干细胞分化的诱导和对细胞迁移能力的影响.结果:三种底物均能诱导神经干细胞的分化,诱导分化的能力为层黏连蛋白＞多聚鸟氨酸＞鼠尾胶原,多聚鸟氨酸与层黏连蛋白两者联用具有协同叠加效应,而且这些底物也具有介导分化后的神经细胞迁移的能力,各种底物对细胞迁移影响力的大小与其诱导分化的能力相似.结论:多聚鸟氨酸、层黏连蛋白和胶原不同程度上促进神经干细胞分化和迁移.