脊髓脑源性神经营养因子表达水平评估大鼠坐骨神经缺损修复程度

目的研究应用脊髓脑源性神经营养因子(BDNF)表达程度评价大鼠坐骨神经缺损修复情况。方法取大鼠30只,随机分成正常组、自体神经组和硅胶管组,每组10只。自体神经组将剪断的10 mm神经缝合于原处,硅胶管组用10 mm医用硅胶管桥接神经缺损,正常组不手术。饲养4个月,于脊髓腰膨大处取材,切片,免疫荧光染色,荧光显微镜下观察。结果各组脊髓前角内均有BDNF表达,表现为点状红色荧光,均匀弥漫分布,胶质细胞内及周围均有分布,未染出神经元。光密度正常组>自体神经组>硅胶管组。结论应用脊髓脑源性神经营养因子表达程度可以评价周围神经损伤的修复情况。     

脑梗死患者外周血淋巴细胞神经营养因子及受体mRNA和相关蛋白质产物的表达

目的 研究脑梗死后外周血淋巴细胞神经营养因子（NGF、BDNF、NT-3）及受体mRNA和相关蛋白质表达变化。方法 用RT-PCR和Western blot技术观测外周血淋巴细胞神经营养因子及受体mRNA和相关蛋白质的动态变化。结果急性期外周血淋巴细胞NT-3及受体mRNA和相关蛋白质表达较对照组明显减少（P〈0．01）,恢复期逐渐上升;急性期NGF与BDNF及其受体mRNA和相关蛋白质表达较对照组显著增加（P〈0．01）,恢复期无明显表达。结论 脑梗死急性期时NGF与BDNF及其受体mRNA和相关蛋白质表达增加,而NT-3及受体mRNA和相关蛋白质表达在缺血初期下降,提示缺血早期NT-3可增加神经元死亡。     

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。     

神经营养因子介导人胚胎干细胞存活

作为多能性群体的人胚胎干细胞（hES），其生长要求在存活、增殖和自我更新信号之间保持平衡。本研究论证了hES细胞能表达介导抗凋亡信号的原肌凝蛋白相关性激酶（TRK）家族的受体。发现3种TRK配体——脑源性神经营养因子、神经营养因子3和4是hES细胞的存活因子。向hES细胞培养物中加入神经营养因子，可使其克隆的存活提高36倍。在含有神经营养因子的培养基中，hES细胞仍然为二倍体并保留了完整的发育潜力。在存在神经营养因子的条件下，hES细胞中的TRK受体被磷酸化；抑制TRK受体将导致hES细胞凋亡。hES细胞中神经营养因子的残留活性受磷脂酰肌醇-3-激酶途径而非分裂索激活的蛋白激酶途径介导。神经营养因子能提高hES细胞的存活能力并且可能有助于对它们的操控，据此可开发高通量筛选模型以鉴别负责hES细胞分化的因子。     

神经干细胞移植治疗脊髓损伤的实验性研究进展

脊髓损伤 (spinalcordinjury)后机体历经原发性损伤和继发性损伤的序贯过程 ,造成了灾难性的后果 ,表现为损伤节段平面以下感觉、运动功能的完全丧失和大小便失禁。Grossman[1] 等在脊髓挫伤的动物模型上证实脊髓功能的永久性障碍主要是继发性损伤造成的神经元和神经胶质丧失导致的。因为中枢神经系统自我修复能力有限 ,故而目前唯一可行的方法是通过移植来替代缺失的神经细胞[2 ] 。在急性脊髓损伤的动物模型上 ,通过移植细胞桥接组织 (外周神经、雪旺细胞 )、胚胎中枢神经细胞、分泌神经营养因子 3(NF 3)的成纤维细胞、杂交瘤细胞 (能…     

左乙拉西坦对难治性癫痫幼鼠海马组织中Slit2、srGAP2和GAP-43动态表达的影响

目的:探讨左乙垃西坦对难治性癫痫幼鼠海马组织中神经轴突导向因子2(Axon guidance factor 2,Slit2)、再生修复关键分子2(Key molecules for regeneration and repair 2,srGAP2)、生长相关蛋白43(Growth associated protein4...     

强脑因子抗脑衰老作用的实验研究

对动物脑组织提取物－强脑因子，进行了类神经营养因子抗脑衰老功能的检测。结果表明该因子对人胚胎脑神经培养细胞有促生存活性和促突起生长活性，对成年小鼠的学习能力有一定的程度的改善，并可显著提高老化鼠抗自由基能力，证明强脑因子是具有抗神经系统衰老作用的生物活性制剂。     

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。     

食管鳞癌组织中Netrin-1蛋白及mRNA的表达

目的 检测食管鳞癌组织中神经轴突导向因子(Netrin-1)及mRNA表达.方法 应用免疫组化SP法和原位杂交方法 检测50例食管鳞癌组织及其相应的19例癌旁不典型增生组织和20例正常食管黏膜组织中Netrin-1、mRNA的表达.结果 ①食管鳞癌组织中Netrin-1、mRNA阳性表达率均高于癌旁不典型增生组织和正常食管黏膜组织(P均<0.05).②有淋巴结转移组食管鳞癌组织中Netrin-1、mRNA阳性表达率均高于无淋巴结转移组(P均<0.05).结论 人食管鳞癌组织中Netrin-1蛋白及mRNA均呈高表达,可能参与食管癌的发生、发展.     

丁苯酞对神经干细胞向神经细胞分化的促进作用

目的 研究丁苯酞通过PI3K-AKT信号通路促进神经干细胞向神经细胞分化的作用。方法 分离培养SD大鼠胚胎脑皮质层的神经干细胞,采用分化专用完全培养基培养后,通过光镜下观察和Nestin蛋白免疫荧光法进行鉴定。设置对照组和实验组,其中实验组添加丁苯酞辅助分化,分为丁苯酞低剂量组（1 μmol·L^-1）、中剂量组（5 μmol·L^-1）和高剂量组（10 μmol·L^-1）,作用24 h后,CCK-8法检测神经干细胞的细胞活力,碱性磷酸酶染色法检测神经干细胞的分化能力,免疫荧光法检测神经细胞分化程度,RT-qPCR法检测转录因子SOX2、PAX6和NeuN的mRNA表达水平,Western-blot及RT-qPCR检测PI3K-AKT信号通路的表达。结果 丁苯酞干预后神经干细胞活力提高,分化能力提高,碱性磷酸酶试验呈强阳性,转录因子SOX2、PAX6和NeuN的mRNA表达水平增高,PI3K-AKT信号通路蛋白和mRNA表达水平均增高,且呈现剂量依赖性。结论 丁苯酞可以促进神经干细胞向神经细胞分化,其作用机制可能与PI3K-AKT的激活有关。     

大麻酚类受体—1在脊髓,背角,背根神经节和周围神经的分布

AIM: The localization of CB1 receptors in the spinal cord, spinal roots, dorsal root ganglion (DRG), and peripheral nerve of the rat was determined. METHODS: We studied the distribution of CB1 cannabinoid receptors by immunohistochemistry using an antibody raised against the N-terminal of the receptor. RESULTS: The spinal cord showed numerous transverse fibers labelled for CB1 receptors throughout and concentrated in the dorsal horn. Lightly-stained cells were observed throughout the spinal cord gray matter. The DRG also showed cells and fibers labelled for CB1 receptors. Labelled fibers were observed in both dorsal and ventral roots as well as in peripheral nerves. CONCLUSION: The presence of CB1 receptors in the DRG, the dorsal root, and the dorsal horn is in accordance with the analgesic effects of cannabinoids. The presence of labelled cells and fibers in the ventral horn and ventral root provides a substrate for cannabinoid-induced muscle relaxant and antispastic effects.     

大麻酚类受体-1在脊髓、背角、背根神经节和周围神经的分布(英文)

AIM: The localization of CB1 receptors in the spinalcord, spinal roots, dorsal root ganglion (DRG), andperipheral nerve of the rat was determined.METHODS: We studied the distribution of CB1cannabinoid receptors by immunohistochemistry usingan antibody raised against the N-termina1 of thereceptor. RESULTS: The spinal cord showednumerous transverse fibers labelled for CB1 receptorsthroughout and concentrated in the dorsal horn.Lightly-stained cells were observed throughout thespinal cord gray matter. The DRG also showed cellsand fibers labelled for CB1 receptors. Labelled fiberswere observed in both dorsal and ventral roots as well as     

Specific binding sites for 125I-endothelin-1 in the porcine and human spinal cord.

Specific binding sites for 125I-endothelin-1 (125I-ET-1) in the spinal cord were investigated using quantitative receptor autoradiographic and chemical cross-linking methods. The binding sites were highly concentrated in porcine and human spinal cord areas corresponding anatomically to the dorsal horn (Rexed's laminae I-III), an area around the central canal (lamina X) and the principal part of the intermediolateral nucleus (IMLp). The localization of the binding sites differed from those of 125I-omega-conotoxin GVIA (125I-CgTx) and 125I-Bolton-Hunter substance P (125I-BH-SP), with the exception that the IMLp shared 125I-ET-1 with 125I-CgTx and 125I-BH-SP binding sites. Specific 125I-ET-1 binding sites in the areas examined were characteristically single and of high affinity. There were no differences between the potencies of unlabeled ET family peptides, ET-1, ET-2, ET-3 and sarafotoxin S6b at inhibiting 125I-ET-1 binding to the areas. Chemical cross-linking studies showed that 125I-ET-1 and 125I-ET-3 mainly bound to a protein with molecular mass of 43 kDa in the porcine and human thoracic spinal cord membranes. The present finding shows the neuronal significance of this newly discovered peptide in the spinal cord.     

Developmental expression of CAPON and Dexras1 in spinal cord of rats

To study the expression of the carboxy-terminal PSD-95/DLG/ZO-1 ligand of nNOS (CAPON) and Dexras1 mRNA during development in the spinal cord of rats, real-time polymerase chain reaction (Real-time PCR), as a quantitative method, was used to study the developmental expression of CAPOn and Dexras1 mRNA level in the spinal cord. The spatial expression of CAPON and Dexras1 mRNA was examined by a combination of in situ hybridization (ISH) and immunofluorescence. During the development of the spinal cord, CAPON mRNA was expressed in low levels from embryo day 14 to day 18. At postnatal day 1, it reached the peak and was expressed in the part which will become the dorsal horn when mature. It then decreased gradually until postnatal week 12, when it presented in the ventral horn. At embryo day 14, Dexras1 mRNA was expressed at low levels, increased during embryo day 16 to day 18 and peaked at postnatal day 1. Spatiotemporal expression of Dexras1 mRNA was similar to CAPOn as confirmed by correlation analysis and colocalization. CAPOn and neuronal nitric oxide synthase (nNOS) was expressed within the same cells of the dorsal horn at postnatal day 1 but had different subcellular localizations. Co-expression of CAPOn and Dexras1 mRNA in myeloid tissue during development process of rat indicates that the adaptor protein, CAPON may play a probable role in differentiation of neurons, synaptic plasticity and synaptogenesis by regulating nNOS to activate Dexras1. __________ Translated from Chinese Journal of Neuroanatomy, 2007, 23(4): 349–354 [译自: 神经解剖学杂志]     

Upregulation of S100A4 after spinal cord transection in adult rats

AIM: To investigate whether spinal cord transection induces changes of gene expression of S100A4 protein. METHODS: In a spinal cord transection model, S100A4 expression and cellular localization were examined using cDNA microarray, Northern blot, immunohistochemistry, and immunofluorescence double-labeling methods. RESULTS: There was very limited S100A4 mRNA expression in the control spinal cord. However, S100A4 mRNA expression was increased significantly in both the rostral and caudal spinal cord segments adjacent to the injury site. Specifically, S100A4 gene expression was substantially increased at d 2, peaked at d 7 and d 14, and remained high up to 28 d post-injury. During its peak expression, S100A4 protein was localized in astrocytes of the spinal cord within 5 mm from the site of spinal transection. CONCLUSION: Spinal cord transection induces prolonged S100A4 expression at both mRNA and protein levels in areas close to the injury site. Increased expression of S100A4 in astrocytes after spinal cor     

Role of reactive oxygen species and spinal cord apoptotic genes in the development of neuropathic pain.

A mouse model of neuropathic pain consisting of chronic constriction injury (CCI) of the sciatic nerve was used to examine the involvement of reactive oxygen species (ROS) in early spinal cord pro-apoptotic gene over-expression during the development of neuropathic pain. RT-PCR analysis showed increased expression of bax, apoptotic protease-activating factor-1 (apaf-1), and caspase-9 in the dorsal horn spinal cord 3 days after chronic constriction injury of sciatic nerve. Consistent with biomolecular data, a marked increase in TUNEL-positive and caspase-3 active form was observed by 3 days CCI. Administration of phenyl-N-tert-butylnitrone (PBN), a potent ROS scavenger, reduced the development of thermal hyperalgesia and mechanical allodynia at 1 and 3 days post-CCI, and decreased the mRNA levels of bax, apaf-1, and caspase-9. PBN also reduced apoptotic and active Caspase-3 positive profiles in the superficial laminae (I-III) of the spinal cord. This study provides evidence that PBN inhibits over-expression of pro-apoptotic genes and neural apoptosis in the spinal cord dorsal horn induced by early-CCI of the sciatic nerve. These findings suggest that ROS regulate expression of some apoptotic genes which might play a role in the onset of neuropathic pain.     

坐骨神经慢性压迫损伤性疼痛与脊髓背角P物质关系研究

目的观察大鼠坐骨神经慢性压迫性损伤（CCI）后脊髓背角P物质表达的变化,探讨P物质在疼痛发生机制中的作用。方法SD雄性大鼠60只,随机分为：A组：CCI组（30只）;B组：对照组（30只）。术前及术后3、7、14、28 d分别测定大鼠热痛阈值、机械痛阈值和行为学评分。术后3、7、14、28d每组取4只,麻醉后用4%多聚甲醛灌注固定,取L4-6段脊髓,以备免疫组化,测定SP的变化。结果所有CCI动物从术后第3天起,出现明显的疼痛行为学改变和热痛阈值、机械痛阈值的降低,与对照组比较差异有统计学意义（P〈0.05或P〈0.01）。免疫组织化学结果表明,A组术后术侧明显高于B组（P〈0.05或P〈0.01）;A组术侧明显高于健侧（P〈0.05或P〈0.01）;而B组仅在第4天术侧高于健侧（P〈0.05）。结论慢性坐骨神经损伤后,脊髓背角SP的表达增加,而且表达增加与CCI大鼠的痛觉过敏、行为变化在时相上基本一致,说明CCI大鼠痛觉过敏与脊髓背角SP的表达增加有关。     

促皮质素对甲醛引起大鼠脊髓内生长抑素的影响

目的：研究促皮质素（Ｃｏｒ）对甲醛引起大鼠脊髓内生长抑素及其合成的影响,方法：采用免疫组织化学,免疫组织化学双重染色法和原位杂交技术。结果：足底注射甲醛使大鼠脊髓背角内ＦＬＩ,ＳｏｍＬＩ,Ｓｏｍ－ＬＩ／ＦＬＩ和ＰＰＳ－ｍＲＮＡ神经元数均对较对照组显著增多,单独ｉｐＣｏｒ对脊髓背角ＦＬＩ和Ｓｏｍ－ＬＩ无明显影响。但是,ｉｐＣｏｒ显著抑制甲醛引起的背角内ＦＬＩ,Ｓｏｍ－ＬＩ,Ｓｏｍ－ＬＩ／ＦＬＩ和ＰＰ     

地塞米松对哮喘豚鼠肺及内脏感觉传入部位IL-1β表达的抑制作用

目的探讨地塞米松对哮喘豚鼠肺及内脏感觉传入系统(C7T5脊神经节及对应的脊髓后角)IL-1β表达的抑制作用。方法以卵蛋白致敏豚鼠制作哮喘模型,免疫组织化学检测各组豚鼠C7T5脊神经节及对应的脊髓后角IL-1β的表达,Westernblot免疫印迹检测各组豚鼠肺、C7T5脊神经节及对应的脊髓后角IL-1β的蛋白表达变化。结果哮喘组豚鼠IL-1β在肺及内脏感觉传入系统表达明显高于对照组(P<0.01),而地塞米松组则明显低于哮喘组(P<0.01)。结论IL-1β可能参与哮喘的发病过程,地塞米松抑制哮喘豚鼠IL-1β的表达可能是激素治疗哮喘的机制之一。     

Autoradiographic evidence for dopaminergic innervation in guinea pig spinal cord

In studies of the localization of the dopaminergic nerve terminals in the cervical cord of guinea pig, autoradiographic analysis of the spinal cord loaded with [3H]dopamine [( 3H]DA) was done under conditions that prevented the nonspecific uptake of [3H]DA. There was specific labeling in the gray matter and a high density of [3H]DA was present in the dorsal horn (DH). Moderate labeling was observed in the neuropil in the vicinity of the central canal. There were grain concentrations in close approximation to the cell bodies of numerous neurons in the DH and to the cell bodies of a few of the motoneurons in the ventral horn (VH). These dopaminergic terminals are possibly linked to sensory transmission and somatic motor function.