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Involvement of platelet-activating factor (PAF) in glutamate neurotoxicity in neuronal sultures

血小板活化因子（PAF）参与神经元缝谷氨酸神经毒性

基本信息

批准号：
07671504
负责人：
HIRASHIMA Yutaka
金额：
$ 1.41万
依托单位：
TOYAMA MEDICAL AND PHARMACEUTICAL UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671504/
关键词：
hypoxia glutamate platelet-activating factor neurotoxicity hippocampal CA1 cultured neuronal cells 低酸素症

项目摘要

To clarify the role platelet-activating factor (PAF) in glutamate neurotoxicity, in vitro experiments using primary neuronal cultures were performed. The anti-PAF immunoglobulin-G (aPAF-IgG) and the three PAF receptor antagonists (BN52021, CV6209, and E5880) were tested for their neuroprotective activity in primary neuronal cultures isolated from embryonic rat cerebral cortex. The cultured neurons were exposed to glutamate (1mM) for 60min. Twenty-four hours after this exposure, aPAF-1gG demonstrated evidence of protective effects against neuronal damage in a dose-dependent manner. Protective effects aiso were observed in cultures treated with the three PAF antagonists (P<0.05 at 1 mug/ml aPAF P<0.01 at 100 muM BN52021, P<0.05 at 10 nM CV6209 and P<0.01 at 10 nM E5880). The Fura-2 assay was used to estimate whether low dosages of exogenous PAF affect cultured neurons. The cultured neurons were loaded with Fura-2/AM.After preincubation for 120 min, the Fura-2-loaded neurons were exposed to various concentrations of PAF for 60 min. By measuring the fluorescent intensity of the medium as representing the amount of Fura-2-released from damaged neurons, we detected an increased release of Fura-2, even at low doses of PAF (P<0.01 at 10 nM PAF). We further studied PAF production by neurons in response to glutamate. The level of PAF measured in the medium exposed to glutamate was significantly higher than the level in the medium unexposed to glutamate (P<0.05). Our results suggest an important role of PAF in glutamate neurotoxicity.

为了阐明血小板活化因子（PAF）在谷氨酸神经毒性中的作用，使用原代神经元培养物进行体外实验。抗PAF免疫球蛋白-G（aPAF-IgG）和三种PAF受体拮抗剂（BN 52021、CV 6209和E5880）在从胚胎大鼠大脑皮层分离的原代神经元培养物中测试其神经保护活性。将培养的神经元暴露于谷氨酸（ImM）60 min。暴露24小时后，aPAF-1gG显示出剂量依赖性的神经元损伤保护作用。在用三种PAF拮抗剂处理的培养物中也观察到保护作用（在1 μ g/ml aPAF时P<0.05，在100 μ M BN 52021时P <0.01，在10 nM CV 6209时P<0.05，在10 nM E5880时P<0.01）。Fura-2检测用于评估低剂量外源性PAF是否影响培养的神经元。将培养的神经元负载Fura-2/AM，预孵育120 min后，将负载Fura-2的神经元暴露于不同浓度的PAF中60 min。通过测量培养基的荧光强度来代表受损神经元释放的Fura-2的量，我们检测到即使在低剂量的PAF下Fura-2的释放也增加（P<0.01，在10 nM PAF下）。我们进一步研究了神经元对谷氨酸的反应。谷氨酸组PAF水平显著高于非谷氨酸组（P<0.05）。我们的研究结果表明PAF在谷氨酸神经毒性中起重要作用。