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本研究通过体外培养大鼠海马神经元建立缺糖缺氧损伤(OGD)模型,观察丹皮酚对神经元的保护作用及其对NMDA受体结合活性的抑制作用。原代培养的神经元于建立OGD模型前5min分别加入丹皮酚及MK-801并观察其作用。OGD模型建立后神经元存活率显著降低,氨基酸分析仪测得的海马神经元细胞外液兴奋性氨基酸(EAA)浓度显著升高,且海马神经元NMDA受体结合活性及NMDA受体NR1亚基mRNA表达显著增强。而经过丹皮酚及MK-801处理后,倒置相差显微镜下观察神经元胞体损伤明显减轻,丹皮酚及MK-801显著降低了细胞死亡率(P<0.05)及细胞外液EAA的升高(P<0.05),并可抑制NMDA受体结合活性的增高及减弱NR1亚基受体mRNA的上调(P<0.05)。实验结果表明,丹皮酚对离体培养的大鼠海马神经元OGD损伤具有保护作用,其作用机制可能与抑制兴奋性氨基酸升高及影响NMDA受体有关。
In this study, we established the model of hypoglycemic-hypoxia injury (OGD) by culturing rat hippocampal neurons in vitro and observing the protective effect of paeonol on neurons and its inhibitory effect on the NMDA receptor binding activity. Primary cultured neurons were added paeonol and MK-801 5 minutes before the establishment of OGD model and observed its role. The survival rate of neurons in OGD model was significantly decreased. The concentration of excitatory amino acid (EAA) in hippocampal neurons increased significantly after amino acid analyzer was measured, and NMDA receptor binding activity and NMDA receptor NR1 subunit mRNA expression was significantly enhanced. After treated with paeonol and MK-801, the damage of neuronal cell bodies was observed under inverted phase contrast microscope. Paeonol and MK-801 significantly reduced cell death rate (P <0.05) and increased extracellular EAA (P <0.05), and inhibited the increase of NMDA receptor binding activity and the up-regulation of NR1 subunit mRNA (P <0.05). The experimental results show that paeonol can protect the hippocampal neurons from injury of OGD in vitro. The mechanism may be related to the inhibition of excitatory amino acids and the influence of NMDA receptors.