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目的:有研究发现谷氨酸受体(GluR)与难治性癫的发作相关,故研究GluR亚单位抗体(GluR3抗体)对海马神经元的损伤作用。方法:取SD新生鼠海马皮质组织,以此培养的原代海马神经元作为靶细胞。抗体在不同的作用时间(20和40h)和稀释倍数下,通过细胞存活率(MTT法)和乳酸脱氢酶(LDH)释放量来测定抗体对神经元的毒性;TUNEL染色法测定神经元的凋亡。结果:GluR3抗体对神经元具有细胞毒性和凋亡作用,并与抗体的滴度和作用时间呈剂量依赖性。即使在低滴度的情况下(×1000和×2000),此抗体仍可诱导凋亡,且与对照组比较差异有显著统计学意义(P<0.01,P<0.05)。结论:GluR3抗体的致机制可能与其能诱导神经元的损伤有关。
OBJECTIVE: It has been found that the glutamate receptor (GluR) is associated with the onset of refractory epilepsy. Therefore, the effect of GluR subunit antibody (GluR3 antibody) on hippocampal neurons injury was studied. Methods: Hippocampal cortical tissues of SD neonatal rats were taken as primary cultured hippocampal neurons. Antibodies at different time of action (20 and 40h) and dilution times by cell viability (MTT method) and lactate dehydrogenase (LDH) release to determine the amount of antibody to neurons; TUNEL staining assay neurons Apoptosis. RESULTS: GluR3 antibody had cytotoxicity and apoptosis effects on neurons and was dose-dependent with titer and duration of antibody. The antibody could induce apoptosis even in the case of low titer (× 1000 and × 2000), and the difference was statistically significant compared with the control group (P <0.01, P <0.05). Conclusion: The mechanism of GluR3 antibody may be related to its ability to induce neuronal damage.