双嘧达莫可能通过Trk受体促进神经干细胞增殖

目的:探讨双嘧达莫促进体外培养神经干细胞增殖的机制。方法:取新生小鼠大脑进行神经干细胞原代培养,以2.5μmol·L~(-1)双嘧达莫组作为阳性对照组,分别加入腺苷受体拮抗剂CGS15943和Trk受体拮抗剂K252a,通过观察神经球形态、神经球生成数目、MTT法定量,初步探讨双嘧达莫促神经干细胞增殖的机制。结果:K252a 双嘧达莫组神经球数量和细胞活性均明显低于双嘧达莫组(P<0,01),而CGS15943 双嘧达莫组与双嘧达莫组相比差异无统计学意义。结论:双嘧达莫的促神经干细胞增殖作用可能是通过Trk受体介导的。     

姜黄素抑制人神经胶质瘤U251细胞增殖的机制

目的 探讨姜黄素对人神经胶质瘤U251细胞增殖的抑制作用及其相关机制.方法 应用0、10、20、40、60、80 μmol/L的姜黄素处理人神经胶质瘤U251细胞,检测细胞增殖、细胞周期、细胞凋亡及葡萄糖转运蛋白-3(GLUT-3)表达情况.结果 10、20、40、60、80 μmol/L姜黄素作用于U251细胞吸光度(OD)值低于0μmol/L组(P＜0.05).姜黄素组作用72 h时人神经胶质瘤U251细胞G1期细胞比例少于对照组,G2/M期细胞比例多于对照组(P＜0.05).两组细胞凋亡率比较差异无统计学意义(P＞0.05).姜黄素组GLUT-3表达水平低于对照组(P＜0.05).结论 姜黄素可以抑制人神经胶质瘤U251细胞增殖,其作用可能是通过下调GLUT-3的表达水平来实现的.     

姜黄素对β-淀粉样蛋白损伤神经干细胞增殖和凋亡的影响

目的探讨姜黄素对β-淀粉样蛋白损伤神经干细胞(NSC)增殖和凋亡的影响。方法选取NSC分为对照组、姜黄素组、Aβ组和观察组,其中姜黄素组、Aβ组和观察组分别加入终浓度为5μmol/L的姜黄素、0.5μmol/L的Aβ+DMSO(10μL)、5μmol/L的姜黄素+0.5μmol/L的Aβ,对照组加入DMSO(10μL)。使用姜黄素治疗NSC,通过细胞计数法计算NSC的增殖速度,噻唑蓝(MTT)测定NSC活性,逆转录。聚合酶链反应(RTPCR)测定NSC中Nestin mRNA及凋亡相关基因Caspase3 mRNA表达,免疫组化测定NSC向神经元和星形胶质细胞的分化潜能。结果与对照组比较,姜黄素组NSC的细胞活力、Nestin mRNA表达明显增加,Caspase3mRNA表达明显降低(P 0.05或P 0.01),而Aβ组和观察组NSC活力、Nestin mRNA表达和Tuj-1、GFAP明显降低,Caspase3 mRNA表达明显增加,差异有统计学意义(P 0.05或P 0.01)。与Aβ组比较,观察组NSCs细胞活力、Nestin mRNA表达和Tuj-1、GFAP明显增加,Caspase3 mRNA表达明显降低,差异有统计学意义(P 0.05或P 0.01)。结论姜黄素促进NSC增殖,促进NSC向神经元分化,逆转Aβ对NSC的损伤可能是由于抑制了凋亡基因表达。     

BVT-14225通过抑制11β-羟基类固醇脱氢酶1还原活性促进神经干细胞增殖和迁移

目的探讨BVT-14225(BVT)通过抑制11β-羟基类固醇脱氢酶1(11β-HSD1)的还原活性对神经干细胞(NSC)增殖、分化和迁移的影响。方法取孕15 d SD胎大鼠的大脑皮质,分离培养原代NSC,采用免疫荧光染色法对NSC标志物巢蛋白和性别决定基因相关转录因子2(Sox2)进行染色,鉴定原代细胞;逆转录聚合酶链反应及核酸凝胶电泳法检测11β-HSD1 mRNA在NSC上的表达。细胞随机分为5组:细胞对照组(培养48 h)、溶剂对照组(0.1%DMSO孵育48 h)、DHC模型组(DHC 10.0μmol·L^-1处理48 h)、BVT对照组(BVT 10.0μmol·L^-1处理48 h)和BVT干预组(BVT 10μmol·L^-1处理1 h后再给予DHC10.0μmol·L^-1,继续处理48 h)。5-乙炔基-2’脱氧尿嘧啶核苷(EdU)细胞增殖试剂盒检测NSC的增殖能力;乳酸脱氢酶(LDH)释放实验检测细胞毒性;超高效液相色谱-质谱(UPLC-MS)检测11β-HSD1酶的还原活性;Wes...     

孤儿核受体Tailless样蛋白在神经干细胞增殖中的作用

孤儿核受体Tailless样蛋白（TLX）是一种类似于核蛋白的转录因子,最新研究发现其对维持细胞功能可能具有重要影响,尤其在神经干细胞增殖方面发挥非常关键的作用。本文将讨论孤儿核受体TLX在神经干细胞增殖过程中的作用,并分析TLX调控神经干细胞增殖作用的分子机制,期望能为治疗神经退行性疾病提供一个潜在的药物新靶点。     

慢病毒介导酪氨酸激酶受体 A过表达载体促进神经干细胞增殖和分化的实验研究

目的探究酪氨酸激酶受体 A(Trk A)过表达载体对神经干细胞增殖和分化能力的影响。方法取 SD大鼠海马组织进行提取神经干细胞,采用免疫荧光的方法对神经球进行鉴定,利用 siRNA技术构建 Trk A过表达载体,将培养的神经干细胞分成三组,空白对照组为未做任何处理的细胞组,阴性对照组是经空白载体的慢病毒转染的细胞组,实验组是由 Trk A的过表达载体构建的慢病毒转染细胞组。逆转录 PCR(RT?PCR)检测 Trk A和神经干细胞分化标志基因 Tuj1,GFAP和 CNPase的表达, CCK?8试剂盒检测各组细胞的增殖率。结果免疫荧光染色显示神经干细胞细胞球 Nestin(红色荧光), Brud染色(绿色荧光)阳性,且占细胞量的 90%以上;荧光双标染色(棕色荧光)显示双染细胞量占总细胞量的 90%以上,明培养的细胞是神经干细胞。慢病毒滴度的检测结果: Trk A过表达慢病毒载体转染滴度为 3×108 TU/mL。RT?PCR结果说表明,空白对照组(1.09±0.09)和阴性对照组(1.11±0.05)的 Trk A的 mRNA相对表达量比较差异无统计学意义(P＞0.05),实验组的 Trk A的 mRNA相对表达量(7.12±1.32)要明显高于空白对照组和阴性对照组(P＜0.05);神经干细胞分化标志基因 Tuj1,GFAP和 CNPase的表达与 Trk A相同。 CCK?8试剂盒检测结果表明,空白对照组、阴性对照组及实验组的细胞增殖率分别为(87.92±9.21)%、(88.22±9.37)%、(176.01±11.33)%(F＝18.082,P＜0.05)其中空白对照组和阴性对照组的细胞增殖率比较差异无统计学意义(P＞0.05)实验组的细胞增殖率要明显高于空白对,照组和阴性对照组(P＜0.05)。结论 Trk A过表达载体可以促进神经元干细胞向,神经元细胞,星型胶质细胞和少突胶质细胞的分化,并且可以促进神经元干细     

腺苷促进体外培养神经干细胞增殖作用研究

目的研究腺苷促进体外培养神经干细胞（NSCs）增殖作用。方法取新生小鼠大脑进行神经干细胞原代培养,观察神经球生成情况,特异性蛋白质免疫细胞化学染色和BrdU标记鉴定并判断其增殖能力;将腺苷按0.4,2.0,10.0 μmol&#8226;L 13个浓度加入神经干细胞培养基,同时设立溶媒对照组和阳性对照组,通过观察神经球形态、神经球生成数目及 MTT法定量比较,分析不同浓度腺苷对神经干细胞分裂增殖的影响。结果培养物中有大量神经球生成,神经干细胞特异性蛋白质免疫细胞化学染色呈阳性,BrdU标记亦呈阳性反应,所培养的细胞为神经干细胞,具有增殖能力;腺苷中、高浓度组神经球数目、MTT比色结果明显高于溶媒对照组。结论腺苷具有促进NSCs增殖作用。     

糖皮质激素受体与新型糖皮质激素受体调节剂的研究进展

外源性糖皮质激素（GCs）在临床上广泛用于治疗炎症性疾病,其机制主要通过糖皮质激素受体（GR）发挥作用。但长期大量的使用GCs引起的不良反应使得GCs的使用受到限制,学术界正不断努力寻找更安全的GR调节剂。通过对GR的作用机制及长期使用GCs导致的不良反应进行介绍,同时,总结了近年来更安全的新型GR调节剂的研究进展,以期为新型GR配体的研发提供参考。     

姜黄素对肿瘤细胞增殖的影响

目的观察姜黄素在体外对多种常见肿瘤细胞增殖的影响。方法分别用0、1、2、5、10、20、50、100μmol·L^-1姜黄素处理人乳腺癌细胞（MCF-7）、人子宫颈癌细胞（Hela）、人肺癌细胞（A549）、人肝癌细胞（HepG-2）24、48、72 h,采用MTT法检测细胞增殖抑制率,并计算72 h时的半数抑制浓度（50%concentration of inhibition,IC50）。结果姜黄素在体外对上述4种细胞增殖均有抑制作用,且呈浓度、时间依赖性。结论姜黄素具有抑制多种肿瘤细胞增殖的作用。     

糖皮质激素对人卵巢癌细胞系3AO增殖分化及对糖皮质激素受体表达的影响

目的：观察不同浓度地塞米松（Dex)对人卵巢癌细胞系（3AO）增殖及分化的影响及其对糖皮质激素受体（GR）的调节作用。方法：以不同浓度Dex处理3AO细胞,采用活细胞计数法观察细胞增殖,用氨基安替比林法测定碱性磷酸酶（AKP）活性;用酶联免疫法测定细胞标志抗原CA125水平的变化;用放射配体结合法测定GR的表达。结果：Dex对3AO细胞的增殖有抑制作用,同时伴有细胞形态的变化及AKP活性增高和CA125的表达下降。Dex对3AO细胞AKP活性的诱导作用可被RU486所阻断。在3AO细胞中存在糖皮质激素受体（GR）,Dex对GR结合活性有下调作用。结论：Dex对3AO细胞有增殖抑制和诱导分化作用,这种作用是通过GR来介导的。     

Diclofenac inhibits proliferation and differentiation of neural stem cells

Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used in clinical situations as anti-inflammatory, analgesic and antipyretic drugs. However, it is still unknown whether NSAIDs have effects on the development of the central nervous system. In the present study, we investigated the effects of NSAIDs on neural stem cell (NSC) proliferation and differentiation into neurons. In contrast to aspirin, naproxen, indomethacin and ibuprofen, treatment with diclofenac (10 microM) for 2 days induced the death of NSCs in a concentration-dependent manner. Diclofenac also inhibited the proliferation of NSCs and their differentiation into neurons. Treatment with diclofenac resulted in nuclear condensation (a morphological change due to apoptosis of NSCs) 24hr after the treatment and activated caspase-3 after 6 hr, indicating that diclofenac may cause apoptosis of neuronal cells via activation of the caspase cascade. These results suggest that diclofenac may affect the development of the central nervous system.     

Melatonin modulates rat myotube-acetylcholine receptors by inhibiting calmodulin

Melatonin, the pineal gland hormone, modulates alpha-bungarotoxin sensitive nicotinic acetylcholine receptors in sympathetic nerve terminals, cerebellum and chick retina imposing a diurnal variation in functional responses [Markus, R.P., Zago, W.M., Carneiro, R.C., 1996. Melatonin modulation of presynaptic nicotinic acetylcholine receptors in the rat vas deferens. J. Pharmacol. Exp. Ther. 279, 18-22; Markus, R.P., Santos, J.M., Zago, W., Reno, L.A., 2003. Melatonin nocturnal surge modulates nicotinic receptors and nicotine-induced [3HI] glutamate release in rat cerebellum slices. J. Pharmacol. Exp. Ther. 305, 525-530; Sampaio, L.F.S., Hamassaki-Britto, D.E., Markus, R.P., 2005. Influence of melatonin on the development of functional nicotinic acetylcholine receptors in cultured chick retinal cells. Braz. J. Med. Biol. Res. 38, 603-613]. Here we show that in rat myotubes forskolin and melatonin reduced the number of nicotinic acetylcholine receptors expressed in plasma membrane. In addition, these cells expressed melatonin MT1 receptors, which are known to be coupled to G(i)-protein. However, the pharmacological profile of melatonin analogs regarding the reduction in cyclic AMP accumulation and number of nicotinic acetylcholine receptors did not point to a mechanism mediated by activation of G(i)-protein coupled receptors. On the other hand, calmidazolium, a classical inhibitor of calmodulin, reduced in a similar manner both effects. Considering that one isoform of adenylyl cyclase present in rat myotubes is regulated by Ca2+/calmodulin, we propose that melatonin modulates the number of nicotinic acetylcholine receptors via reduction in cyclic AMP accumulation.     

胎鼠神经干细胞培养方法的建立及药物对干细胞增殖的影响

目的建立神经干细胞培养模型，观察药物对其增殖能力的影响。方法建立大鼠胎脑神经干细胞的培养方法，并用MTT法和3H-胸腺嘧啶核苷参入法观察黄皮酰胺、丹酚酸A及人参皂苷Rg1等对第2代神经球细胞状态的影响。结果免疫细胞化学结果显示，培养的神经细胞具备干细胞的基本特性；MTT和液闪结果则表明，上述3种药物可不同程度地影响神经干细胞的存活率和/或增殖活性。结论本实验所建立的胚胎大鼠神经干细胞培养模型可以观察到一些有脑保护作用的药物对干细胞存活和增殖有一定影响。     

Growth-stimulating effect of kallikrein on rat neural stem cells

Tissue kallikrein is expressed in many species and is widely distributed throughout the body, including the brain. In general, this protease is well known to release the vasoactive peptide, kinin, from kininogen. We report here that kallikrein has a prominent growth-stimulating effect on neural stem cells prepared from the brains of prenatal rats. This growth-stimulating effect was suppressed by antiserum against rat tissue kallikrein. Since bradykinin B2-receptor antagonist, Hoe140, did not suppress the growth-stimulating effect, kallikrein-mediated kinin release does not appear to be involved in this effect. Thus, our data suggest a new physiological function of kallikrein, the growth of neural stem cells. Such involvement would suggest that kallikrein is not only potentially involved in an important function of brain development, but also available for studies on regenerative medicine for neurons.     

Regulation effect of nardosinone on the proliferation and functioning of neural stem cells

The purpose of this study was to investigate the effect of nardosinone(Nar) on the proliferation,migration and differentiation of mouse neural stem cells(NSC).The NSC were isolated from mice embryos at gestational day E14 and cultured using the neurosphere culture system.Nar or vehicle was added to the culture media at the start of culture.Cell viability and proliferation were measured with MTT and BrdU incorporation assays respectively.Cell cycle was analyzed by flow cytometric cell sorting.The NSC migration and differentiation were monitored with immune-fluorescence techniques.ERK/CREB signaling pathway was detected with Western blotting analysis.The results demonstrate that after 3 d incubation with the cells,Nar 50 μmol·L-1 increased the viability,BrdU incorporation,and the proportion of cells in S and G2 phase(P<0.05).Exposure to Nar for 24 h promoted cell migration in a dose-dependent manner.Quantitative analyses revealed Nar 50 μmol·L-1 and 25 μmol·L-1 increased the proportion of neurons and oligodendrocytes differentiated from NSC after 7 d and 14 d(P<0.05).Nar,co-incubation with the NSC in proliferation media for 3 d,increased the level of phosphorylated ERK1/2(p-ERK1/2) and phosphorylated CREB(p-CREB) in NSC(P<0.05) without altering the expression of ERK1/2 and CREB.Nar 50 μmol·L-1 and 25 μmol·L-1 also increased the level of p-ERK1/2 and p-CREB in NSC after 7 d co-incubation in differentiation media,however,only increased p-ERK1/2 was observed after 14 d treatment.Together,these findings suggest that Nar promotes the proliferation and migration of NSC,and influence the preference of NSC differentiation.These effects may relate to the action on ERK/CREB signaling pathway.     

人胚胎神经干细胞体外培养及其增殖与分化的研究

目的 建立神经干细胞分离、培养及分化的鉴定技术,观察神经干细胞增殖、分化的特点。方法 从人胚胎海马区分离神经干细胞,采用无血清培养基,进行体外扩增培养、传代。采用免疫细胞化学法鉴定神经干细胞和分化的神经细胞;利用流式细胞仪和细胞生长曲线检测神经干细胞的增殖能力。结果 从人胚胎脑海马区分离的细胞具有增殖和多向分化潜能,可进行传代培养,获得的细胞团中大部分为nestin表达阳性细胞。贴壁分化后可以出现NSE、GFAP表达阳性的细胞。结论 用上述方法分离培养的细胞能表达nestin蛋白,具有自我更新和增殖能力,并具有向神经元、星形胶质细胞分化的潜能,具备神经干细胞的特征,可用于细胞移植等相关研究。     

香丹注射液对大鼠骨髓间充质干细胞的增殖作用

目的 研究香丹注射液对大鼠骨髓间充质干细胞(MSC)增殖的影响.方法 肌注香丹注射液和5-溴尿嘧啶核苷(Brdu);体外培养MSC,用四甲基偶氮唑法检测香丹注射液促进大鼠MSC的增殖作用,免疫组化检测MSC细胞的Brdu标记阳性率.结果 3种高浓度的药液对MSC的增殖与对照组比较有显著性差异;停药后3-7 d,香丹组Brdu标记阳性细胞率与生理盐水组比较有显著性差异.结论 香丹注射液有促进MSC增殖的作用.