PURIFICATION AND CHARACTERZATION OF GLIA-DERIVED NEUROPROTECTIVE FACTORS

胶质细胞源性神经保护因子的纯化和表征

• 批准号: 10680745
• 负责人: TANAKA Junya
• 金额: $ 2.05万
• 依托单位: EHIME UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10680745/
• 关键词: MICROGLIA; ASTROCYTES; NEURON; APOPTOSIS; INHIBITION OF CELL DEATH

We have found the followings regarding the neuroprotective effects in this project.1) Neuroprotective effects of microglial cells. We found that cerebrocortical primary neurons died by apoptosis by incubating cells with sodium nitroprusside (SNP), a nitric oxide (NO)-donor compound, By the use of this apoptosis model, we investigated the effect of microglial cells on neuronal survival. As a result, the NO-induced neuronal apoptosis was completely prevented by microglial cells that were cocultured with neurons. Since the neuronal apoptosis was also largely inhibited by microglial conditioned medium, microglia-derived anti-apoptotic factor may be a soluble-factor(s). Furthermore, the conditioned medium was effective when it was applied after exposure to NO. Among a variety of known cytokines and growth factors that are produced by microglial cells, only tumor necrosis factor a significantly rescued neurons against NO-induced damage. 2) Neuroprotective effects of astrocytes. We investigated to determine whether astrocytes could rescue primary cerebrocortical neurons from reactive oxygen species (ROS)-induced apoptosis. By adding SNP, 3-morpholinosydononimine (SIN-1) or mixture of FeSO4 and ascorbic acid, NO, superoxide anion or hydroxyl radicals were introduced into culture plates of neurons. When astrocytes were cocultured with neurons, ROS-induced neuronal death was completely prevented. However, meningeal fibroblasts could not rescue neurons. When neurons were cultured on astrocytes-derived extracellular matrix (ECM), neuronal apoptosis was mostly prevented. The astrocyte-derived ECM contained laminin and fibronectin, and purified laminin and fibronectin significantly protected neurons against oxidative stress.

我们发现了该项目中有关神经保护作用的以下内容。1）小胶质细胞的神经保护作用。我们发现，脑皮质原代神经元因凋亡而死亡，通过使用这种凋亡模型将细胞与硝化钠（SNP）（SNP）（SNP）（SNP）（SNP）孵育，一种氧化氧化物（NO） - 浓度化合物，我们研究了小胶质细胞对神经元存活的影响。结果，与神经元共培养的小胶质细胞完全阻止了无诱导的神经元凋亡。由于小胶质细胞调节培养基也很大程度上抑制了神经元凋亡，因此小胶质细胞衍生的抗凋亡因子可能是可溶性因子（S）。此外，在暴露于NO后将其应用时，条件培养基是有效的。在小胶质细胞产生的各种已知的细胞因子和生长因子中，只有肿瘤坏死因素可显着拯救神经元，以抗非诱导的损伤。 2）星形胶质细胞的神经保护作用。我们研究以确定星形胶质细胞是否可以从活性氧（ROS）诱导的细胞凋亡中挽救原发性脑皮质神经元。通过添加SNP，3-多酚辛二烯酰胺（SIN-1）或FESO4和抗坏血酸的混合物，将NO，超氧化物阴离子或羟基自由基引入了神经元的培养板中。当星形胶质细胞与神经元共培养时，完全阻止了ROS诱导的神经元死亡。但是，脑膜成纤维细胞无法营救神经元。当神经元在星形胶质细胞衍生的细胞外基质（ECM）上培养时，大多可预防神经元凋亡。星形胶质细胞衍生的ECM含有层粘连蛋白和纤连蛋白，纯化的层粘连蛋白和纤连蛋白显着保护神经元免受氧化应激。

项目成果

Toku K,Tanaka J,et al.: "Microglial cells prevent nitric oxide-induced neuronal apoptosis in vitro"Journal of Neuroscience Research. 53. 415-425 (1999)

Toku K、Tanaka J 等人：“小胶质细胞在体外预防一氧化氮诱导的神经元凋亡”神经科学研究杂志。

Tanaka J,Toku K et al.: "Astrocytes prevent neuronal death induced by reactive oxygen and nitrogen..."Glia. 28. 85-96 (1999)

Tanaka J、Toku K 等人：“星形胶质细胞可防止活性氧和氮诱导的神经元死亡......”神经胶质细胞。

Kazuko Toku,Junya Tanaka,et al.: "Microglial cells prevent nitric oxide-induced neuronal apoptosis in vitro." Journal of Neuroscience Research. 53. 415-425 (1998)

Kazuko Toku、Junya Tanaka 等人：“小胶质细胞在体外可预防一氧化氮诱导的神经元凋亡。”

Keiji Igase,Junya Tanaka et al.: "An 18-mer peptide fragment of prosaposin ameliorates place navigation disability,cortical infarction,and retrograde thalamic degeneration in rats with focal cerebral ischemia." Journal of Cerebral Blood Flow and Metabolis

Keiji Igase、Junya Tanaka 等人：“prosaposin 的 18 聚体肽片段可改善局灶性脑缺血大鼠的位置导航障碍、皮质梗塞和逆行丘脑变性。”

Nakatuka H,Ohta S,Tanaka J,et al.: "Release of cytochrome c from mitochondria to cytosol in gerbil hippocampal..."Brain Research. 849. 216-219 (1999)

Nakatuka H、Ohta S、Tanaka J 等人：“沙鼠海马中细胞色素 c 从线粒体释放到胞质溶胶......”大脑研究。