脑出血大鼠脑内NGF和BDNF蛋白表达变化的研究

目的观察脑出血大鼠脑组织内神经生长因子(NGF)和脑源性神经营养因子(BDNF)表达的变化,探讨神经营养因子在脑出血后脑损伤的作用。方法SD雄性大鼠随机分为正常组、假手术组和脑出血组,通过自体血脑内注射法建立脑出血动物模型,采用免疫组织化学法检测脑出血后3、6、12、24、48、72h、7d脑组织中NGF和BDNF蛋白表达情况。结果脑出血组6hNGF表达增加,12hBDNF表达增加,均于24h达高峰,与假手术组及正常组相比差异具有显著性(P<0.05)。结论脑出血后可诱导NGF和BDNF蛋白表达增加,有利于对脑出血后损伤神经元的保护和修复。     

骨髓间充质干细胞移植对大鼠脊髓损伤后再修复作用的研究

目的探讨间充质干细胞（MSCS）移植对大鼠脊髓损伤后脑源性神经营养因子（BDNF）表达的影响。方法选取SD大鼠4只作骨髓间充质细胞的分离与培养，提取MSCS；制做脊髓横断损伤模型，细胞移植组24只，PBS（磷酸盐缓冲液）液组24只，空白对照组12只。于脊髓损伤后第7天，无菌条件下，细胞移植组以微量注射器缓慢注入含MSCS的培养液5μl，磷酸盐缓冲液组5μl，对照组未加任何干预因素。分别于术后7d、14d、28d麻醉下行心脏灌流固定取T10节段脊髓，细胞移植组与磷酸盐缓冲组取出损伤节段的脊髓，空白对照组于同一节段取出相应脊髓。应用免疫组化法观察MSCS移植后大鼠脊髓损伤区BDNF的表达变化。结果脑源性神经营养因子在正常大鼠脊髓组织中有一定表达，间充质干细胞移植术后7d、14d及28d，细胞移植组脑源性神经营养因子均高水平表达，与缓冲液组相比较差别明显，具有统计学意义。结论间充质干细胞在移植后通过上调脑源性神经营养因子的表达从而促进轴突的再生，可能是治疗脊髓损伤的重要机制。     

蒙药新黑苏嘎乌日勒对脑中动脉阻塞-再灌注损伤模型大鼠神经功能损伤的改善作用及机制研究

目的:研究蒙药新黑苏嘎乌日勒对脑中动脉阻塞-再灌注损伤模型大鼠神经功能损伤的改善作用及机制。方法:将80只大鼠随机分为假手术组、模型组、新黑苏嘎乌日勒组(540 mg/kg)、化学药阳性对照组(尼莫地平片,16.12 mg/kg)、中药阳性对照组(银杏叶片,5.18 mg/kg),每组16只;除假手术组外,其余各组大鼠采用Zea-Longa线栓法复制脑中动脉阻塞-再灌注损伤模型;造模成功后,各给药组大鼠灌胃相应药物,假手术组和模型组灌胃等量蒸馏水,连续灌胃14 d。末次给药1 h后,对各组大鼠进行神经功能损伤评分;采用2,3,5-三苯基氯化四氮唑法计算各组大鼠脑梗死率、脑含水率及脑指数;苏木精-伊红染色观察各组大鼠脑组织海马区神经元病理学变化;免疫组化染色法检测各组大鼠脑前额叶脑源性神经营养因子(BDNF)和神经生长因子(NGF)蛋白表达水平。结果:与假手术组比较,模型组大鼠神经功能损伤评分、脑梗死率、脑含水率、脑指数和BDNF、NGF蛋白表达水平均显著升高(P<0.01),脑组织海马区神经元细胞排列松散,形状不规则,部分胞体皱缩,胞浆内尼氏小体减少,核仁消失。与模型组比较,新黑苏嘎乌日勒组、化学药阳性对照组、中药阳性对照组大鼠神经功能损伤评分、脑梗死率、脑含水率、脑指数均显著降低(P<0.05或P<0.01),脑前额叶BDNF、NGF蛋白表达水平均显著升高(P<0.05或P<0.01),脑组织海马区神经元细胞排列整齐紧密,细胞核圆整,细胞质染色均匀。结论:蒙药新黑苏嘎乌日勒对脑中动脉阻塞-再灌注损伤模型大鼠神经功能损伤具有一定的改善作用,其作用机制可能与增加BDNF及NGF蛋白的表达有关。     

类脑救治脑创伤的研究

目的比较和研究不同培养天数类脑移植治疗脑创伤的效果。方法 (1)比较85 d类脑和55 d类脑的细胞数目和数量。(2)使用活检穿孔器在大鼠右侧感觉运动皮质造成机械性损伤,使其出现运动功能障碍。随即在大鼠皮质损伤区域移植55 d类脑和85 d类脑,分为55 d类脑移植组、85 d类脑移植组、创伤组和假手术组。(3)分别在大鼠术后的第7,14,28和56天通过免疫荧光、免疫组化等染色比较各组间神经再生、炎症反应等情况。(4)在手术后的第2,5,7,11,14,21,28,35和42天分别对55 d类脑移植组、创伤组和假手术组大鼠进行m NSS(modified neurological severity scores)评分和平衡木实验,计算各组大鼠的得分情况,评估大鼠术后神经功能恢复情况。结果 (1) 55 d类脑包含更多的神经干细胞,85 d类脑包含更多的成熟神经元,同时具有更多的细胞量。(2)类脑移植可以促进脑创伤大鼠神经再生且不加重炎症反应,55 d类脑移植效果更佳。(3)类脑可以在宿主脑内进一步生长分化,同时上调神经连接蛋白和神经营养因子的表达。(4)类脑移植可以促进脑创伤大鼠运动功能恢复。结论类脑移植可以促进脑创伤大鼠神经再生,上调脑创伤大鼠脑内神经连接蛋白和神经营养因子的表达,改善脑创伤大鼠神经功能,其中55 d类脑移植有更高的细胞存活率和更好的神经保护作用。移植的类脑可以在大鼠脑内存活分化成多种运动皮质区的神经细胞,同时还能进行细胞迁移和与大脑皮质形成血管连接。     

脑缺血大鼠骨髓间质干细胞移植后脑内分布与迁移

目的观察骨髓间质干细胞移植后在脑缺血大鼠脑内的存活、分布及迁移情况。方法Fieoll-Paque分离液梯度离心分离大鼠骨髓间质干细胞（rat bone mesenchymal stem cells，rMSCs），经体外培养扩增并流式细胞术鉴定；线拴法制作大鼠大脑中动脉闭塞（MCAO）脑局灶缺血模型，Hoechst33342标记细胞，分别通过静脉移植及立体定向局部移植到大鼠缺血侧纹状体。经过1、3、5周处死大鼠，取脑组织切片，在倒置荧光显微镜下观察细胞在脑内的存活、分布及迁移情况。结果应用梯度离心方法可以分离得到rMSCs，经过体外扩增可得到足够细胞数量用于移植，流式细胞术检测CD34、CD45表达阴性，CD29、CD90表达阳性。大鼠脑缺血后，无论静脉还是局部移植，rMSCs在脑内均能较长时间存活，移植细胞与宿主有很好组织相容性。静脉移植的细胞集中于脑缺血灶及其周围，缺血灶对侧脑组织移植的细胞较少（P〈0．05）；局部移植的细胞随着时间的推移向缺血灶不断迁移，分布在两侧大脑的差别都具有统计学意义（P〈0．05）。结论rMSCs通过静脉及立体定向两条途径进行移植后，能够在宿主缺血的脑中存活，向损伤部位迁移。rMSCs的这种迁移现象很可能与缺血灶的细胞信号改变有关，在临床上具有很大的潜在价值。     

骨髓基质细胞移植兼用脑源性神经营养因子促进大鼠脊髓损伤修复的实验探究

目的：探讨骨髓基质细胞移植兼用脑源性神经营养因子促进大鼠脊髓损伤修复效果。方法：选取30只Wistar大鼠,按照移植方法分为研究组与对照组,研究组移植采取骨髓基质细胞移植兼用脑源性神经营养因子（BDNF）,对照组单纯移植的骨髓间充质肝细胞（BMSCs）悬液,分析比较两组效果。结果：术后2～8周,两组神经功能评分有统计学差异（P＜0.05）。结论：BMSCs移植能够提高大鼠半横断脊髓结构及功能水平恢复BMSCs与BDNF,联合应用对脊髓损伤修复具有明显协同效果。     

注射用鼠神经生长因子对大鼠缺血脑组织神经生长因子及脑源性神经营养因子表达的影响

目的研究注射用鼠神经生长因子对大鼠缺血脑组织中脑源性神经营养因子(BDNF)和神经生长因子(NGF)表达的影响。方法健康雄性Wistar大鼠,采用改良的Zea-Longa法建立大脑中动脉局灶缺血模型(MCAO)。实验分为模型对照组和注射用鼠神经生长因子组,每组各20只大鼠。注射用鼠神经生长因子组予以腹腔内注射鼠神经生长因子,每天1次,每次9 k U;模型对照组每天注射等量0.9%氯化钠注射液。72 h后行神经功能评分,然后处死,分别取脑梗死周围、海马区和梗死区脑组织作免疫组织化学染色检测BDNF、NGF蛋白表达变化,原位杂交检测BDNF m RNA、NGF m RNA的表达。结果治疗3 d后注射用鼠神经生长因子组神经功能缺损评分低于模型对照组(P<0.05)。注射用鼠神经生长因子治疗组BDNF和NGF蛋白、BDNF m RNA和NGF m RNA在梗死区周围和海马区表达均高于模型对照组,差异有统计学意义(P<0.05),在梗死区差异均无统计学意义(P>0.05)。结论注射用鼠神经生长因子可上调大鼠梗死区周围和海马区缺血脑组织中BDNF和NGF的表达,并可能通过此机制起到保护作用。     

骨髓基质细胞移植抑制缺血心肌纤维化

目的探讨骨髓基质细胞移植改善缺血心脏功能的机制。方法制作F344大鼠心肌梗死模型,将分离培养的骨髓基质细胞移植于心梗周边区（治疗组）,取1、3、5、7、9、15d后缺血心肌切片,通过免疫组织化学染色及免疫荧光染色检测波形蛋白的表达,了解梗死心肌纤维化程度,并应用血液动力学在术后各时间点检测大鼠心脏功能的变化。结果治疗组心功能较同期对照组明显改善。术后7、9、15d治疗组缺血心肌纤维化程度较同期对照组明显减轻。结论骨髓基质细胞移植于缺血心肌后明显改善缺血心脏功能。其机制之一可能是抑制缺血心肌的纤维化,延缓心室重构。     

黄芪总苷对大鼠局灶性脑缺血/再灌注损伤后BDNF、TrkB和p75NTR mRNA表达的影响

目的观察黄芪总苷(astragalosides,AST)对大鼠局灶性脑缺血/再灌注损伤后的神经保护作用,并探讨其作用机制。方法采用大脑中动脉线栓法制备大鼠局灶性脑缺血(MCAO)再灌注模型,观察AST对大鼠缺血/再灌注损伤后1、3、7和14 d的神经功能的影响;采用RT-PCR方法,观察AST对脑组织BDNF和p75NTR mRNA表达的影响;采用Real-time PCR方法,观察AST对脑组织TrkB mRNA表达的影响。结果AST(40 mg.kg-1)在ig后3 d可以明显降低模型大鼠神经功能评分;在3、7和14 d可以提高脑组织中BDNF mRNA的表达;在7 d和14 d可以降低脑组织中p75NTR mRNA的表达,提高TrkB mRNA表达。结论AST对大鼠局灶性脑缺血/再灌注损伤后的神经功能恢复有一定的促进作用,其机制可能与促进BDNF和TrkB mRNA表达、抑制p75NTR mRNA的表达有关。     

BMSCs靶向移植联合EPO法对脊髓损伤组织多因子表达水平的影响

目的 探讨骨髓间充质干细胞(BMSCs)靶向移植联合促红细胞素(EPO)的综合治疗方法对脊髓损伤的修复效果.方法 采用Western blot法检测对照组、模型组、BMSCs治疗组、EPO治疗组和BMSCs联合EPO治疗组大鼠的神经生长因子(NGF)、神经营养因子(NT-3)、脑源性神经营养因子(BDNF)、神经胶质细胞源性神经营养因子(GD-NF)和睫状神经营养因子(CNTF)的表达情况.结果 BMSCs联合EPO综合治疗较BMSCs、EPO单纯治疗更能提高NGF、BDNF、GDNF和CNTF的表达水平.结论 BMSCs联合EPO综合治疗有利于脊髓损伤的修复,具有良好的应用前景.     

APP17肽对心肺复苏大鼠海马神经元NGF、BDNF表达的影响

目的观察心肺复苏术后大鼠海马神经元神经生长因子(nerve growth factor,NGF)、脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)表达的变化,以及APP17肽对NGF、BDNF表达的影响。方法采用窒息法制作大鼠呼吸心跳骤停模型,然后行心肺复苏术(cardiopulmonary re-suscitation,CPR)。♂性Wistar大鼠18只,随机分3组:手术对照组(A组)、心肺复苏组(B组)、心肺复苏+APP17肽组(C组),复苏至自主循环恢复(return of spontaneous circula-tion,ROSC)时静脉注射APP17肽10μg·(300g)-1。应用免疫组化方法观察各组大鼠复苏2h后海马神经元表达NGF、BDNF情况。结果与对照组(A组)相比心肺复苏组(B组)大鼠海马神经元NGF、BDNF表达明显降低(P<0.05);心肺复苏+APP17肽组(C组)NGF、BDNF表达明显增加,高于B组和A组(P<0.01)。结论心肺复苏模型大鼠自主循环恢复2h海马神经元NGF、BDNF表达降低,APP17肽能恢复神经元NGF、BDNF的表达,对缺血、缺氧性神经元损伤有保护作用。     

山茱萸多糖对青霉素致痫幼鼠学习记忆能力及海马组织脑源性神经因子、神经生长因子表达的影响

目的:探讨山茱萸多糖对青霉素致痫幼鼠学习记忆能力及海马组织脑源性神经因子(BDNF)、神经生长因子(NGF)表达的影响。方法:将30只日龄21 d龄幼鼠随机分为生理盐水对照组、青霉素点燃模型组、山茱萸多糖处理组各10只。生理盐水对照组只接受生理盐水灌胃,不造模;山茱萸多糖处理组在造模成功后灌胃0.05 g/mL山茱萸多糖;青霉素点燃模型组造模成功后灌胃等量生理盐水;连续灌胃28 d。采用Morris水迷宫检测各组幼鼠学习记忆能力;RT-PCR法检测海马组织BDNF mRNA、NGF mRNA表达;Western-blot法检测海马组织BDNF与NGF蛋白含量。结果:与生理盐水对照组比较,青霉素点燃模型组幼鼠学习记忆能力明显降低(P<0.01);与模型组比较,山茱萸多糖处理组学习记忆能力明显升高(P<0.01)。与生理盐水对照组比较,青霉素点燃模型组幼鼠BDNF、NGF mRNA和蛋白表达明显降低(P<0.01);与模型组比较,山茱萸多糖处理组BDNF、NGF mRNA和蛋白表达明显升高(P<0.01)。结论:山茱萸多糖具有提高青霉素点燃幼鼠学习记忆能力的作用,其机制可能是通过上调BDNF和NGF基因表达。     

Lithium treatment alters brain concentrations of nerve growth factor,brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor in a rat model of depression

Nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) are proteins involved in neuronal survival and plasticity of dopaminergic, cholinergic and serotonergic neurons in the central nervous system. Since decreased size and impaired function of some neuronal populations may be relevant in depression it has been hypothesized that these molecules may have a functional role in the pathophysiology as well as treatment of depression. Using an animal model of depression, the Flinders Sensitive Line (FSL) rats and their controls, the Flinders Resistant Line (FRL), we investigated the effects of chronic lithium treatment on brain NGF, BDNF and GDNF. Lithium was administered as food supplementation during 6 wk. NGF, BDNF and GDNF measurements were performed by enzyme-linked immunosorbent assay (ELISA). Lithium altered the brain concentrations of neurotrophic factors in the hippocampus, frontal cortex, occipital cortex and striatum. Moreover, the changes were different in the two rat strains. Our data support the notion that neurotrophic factors play a role in depression and in the mechanism of the action of lithium.     

神经生长因子对大鼠坐骨神经损伤的修复作用

目的：观察大鼠坐骨神经横断挫伤后,外源性神经生长因子（NGF）对大鼠周围神经损伤的修复作用。方法：采用大鼠坐骨神经横断致伤方法,实验分为：假手术组、生理盐水对照组、NGF组,每组10只,观察时间为2wk。结果：术后2wk,NGF组的体感诱发电位（SEP）、运动诱发电位（MEP）与生理盐水对照组相比,潜伏期明显缩短（P＜0．05）。经Tarlov评分,NGF组有70％可恢复到4－5分,而生理盐水对照组只有20％恢复到4－5分。结论：NGF对周围神经损伤后有促进神经传导和损伤修复的作用。     

医用三氧对慢性坐骨神经损伤大鼠神经生长因子水平的影响

目的 探讨不同浓度医用三氧(O3)对慢性坐骨神经损伤(chronic constriction injury,CCI)大鼠神经生长因子水平的影响,为临床治疗神经病理性疼痛提供基础科学依据.方法 60只SPF级健康成年雄性SD大鼠随机分为假手术组、CCI组和不同浓度O3(15、30和60μg/ml依序为O3-15组、O3...     

Reduced serum concentrations of nerve growth factor, but not brain-derived neurotrophic factor, in chronic cannabis abusers

Chronic cannabis use produces effects within the central nervous system (CNS) which include deficits in learning and attention tasks and decreased brain volume. Neurotrophins, in particular nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF), are proteins that serve as survival factors for CNS neurons. Deficits in the production and utilization of these proteins can lead to CNS dysfunctions including those associated with cannabis abuse.In this study we measured by enzyme-linked immunosorbent assay (ELISA) the NGF and BDNF serum levels in two groups of subjects: cannabis-dependent patients and healthy subjects. We found that NGF serum levels were significantly reduced in cannabis abusers as compared to healthy subjects.These findings indicate that NGF may have a role in the central action of cannabis and potentially in the neurotoxicity induced by this drug. These data also suggest that chronic cannabis consumption may be a risk factor for developing psychosis among drug users.     

Stimulation of nerve growth factor expression in astrocytes by peroxynitrite

BACKGROUND: Overproduction of nitric oxide (NO) has been recognized as a major mechanism of neurotoxicity. NO reacts with superoxide to generate peroxynitrite, a strong oxidizing and nitrating species. Peroxynitrite is formed in glial cells and degenerating neurons in neuropathological conditions, including amyotrophic lateral sclerosis (ALS). In ALS, motor neurons re-express the p75 neurotrophin receptor (p75NTR) and might become vulnerable to NGF. In the present study, we investigated whether peroxynitrite stimulated nerve growth factor (NGF) expression in spinal cord astrocytes. MATERIALS AND METHODS: Astrocyte monolayers were exposed to peroxynitrite and nitrotyrosine formation was determined by immunofluorescence. mRNA levels for NGF, brain derived neutrophic factor (BDNF) and fibroblast growth factor-2 (FGF-2) were measured by semi-quantitative RT-PCR and NGF release was determined by ELISA. RESULTS AND DISCUSSION: A single exposure to peroxynitrite specifically induced NGF expression and secretion in astrocytes coincident with reactive morphological changes and increased nitrotyrosine immunoreactivity. These results suggest that NGF expression in reactive astrocytes is under the control of oxidative stress.     

Protective effect of nerve growth factor on neurons after traumatic brain injury

The protective effect of nerve growth factor (NGF) on neurons after traumatic brain injury (TBI) was investigated. A brain trauma model of fluid-percussion in rats was established, and 7s NGF was infused continuously in its cerebral ventricle. The activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT), and [Ca2+]i overloading in brain tissues was observed after giving exogenous NGF postinjury. We found that the activity of SOD, GSH-Px, and CAT was markedly higher in NGF-treated group than in the simple trauma group (P < 0.01). Although the level of [Ca2+]i in the NGF-treated group increased, the value was significantly lower than that in the simple trauma and control groups (P < 0.01). These findings suggest that exogenous NGF can (a) increase the activity of the major antioxidant enzymes in brain tissues and attenuate the injuries to neurons induced by oxygen-free radicals, (b) reduce the severe overload of [Ca2+]i and stabilize its homeostasis, and (c) provide clear protective effects on neurons after traumatic brain injury.     

组织型激肽释放酶对脑缺血大鼠神经生长因子的影响

目的探讨组织型激肽释放酶(Tissue kallikrein,TK)治疗大鼠缺血性脑梗死的疗效以及对神经生长因子(NGF)的影响。方法将36只Wistar大鼠随机分配到3组:空白对照组、盐水对照组[NS 2 mL/(kg.d)]、TK组[TK 1 715×10-3U/(kg.d)]。药物在缺血后8 h给予,连续用药3 d。大鼠梗死后1、3及7 d后分别进行神经功能缺失评分,对脑缺血组织NGF进行免疫组化测定。结果治疗3 d后,与盐水对照组及空白对照组相比,TK组神经功能缺损明显减轻(P<0.05),缺血区内NGF表达增多(P<0.05)。结论 TK可以通过增加NGF而治疗大鼠缺血性脑梗死,促进神经细胞再生。