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目的探讨NMDA受体在戊二酸尿症Ⅰ型纹状体神经元兴奋性损伤中的作用。方法利用构建的特异性沉默戊二酰辅酶A脱氢酶(GCDH)慢病毒载体感染原代培养纹状体神经元结合高浓度赖氨酸培养构建GA1细胞模型。Western-Blot检测NMDA受体蛋白表达水平变化。NMDA受体拮抗剂MK-801预处理神经元,再行慢病毒及高浓度赖氨酸干预,MTT检测神经元活性及Hoechst3342检测神经元凋亡情况。结果与对照组比较,实验组的NR2B蛋白表达升高,差异有统计学意义(P<0.001)。实验组、对照组以及MK-801预处理组三组间神经元活性和正常核比例的差异均有统计学意义(P<0.01);两两比较发现,实验组神经元活性和正常核比例较对照组明显下降,MK-801预处理后神经元活性和正常核比例较实验组明显升高,但仍低于对照组,差异均有统计学意义(P<0.05)。结论 NR2B受体介导戊二酸尿症Ⅰ型体内代谢累积物致纹状体神经元损伤。
Objective To investigate the role of NMDA receptors in the excitotoxicity of glutaral aciduria type Ⅰ striatum neurons. Methods The GA1 cell model was constructed by culturing primary cultured striatal neurons with high concentration of lysine and infecting the cultured neurons with glutaraldehyde coenzyme A dehydrogenase (GCDH) lentiviral vector. Western-Blot detection of NMDA receptor protein expression level changes. NMDA receptor antagonist MK-801 was used to pretreat neurons, and then lentivirus and high concentration of lysine were intervened. The activity of neurons was detected by MTT assay and neuron apoptosis was detected by Hoechst3342. Results Compared with the control group, the NR2B protein expression in the experimental group increased significantly (P <0.001). The difference of neuronal activity and normal nuclear ratio between experimental group, control group and MK-801 pretreatment group was statistically significant (P <0.01). Comparing the two groups showed that the neuron activity and normal nuclear ratio Compared with the control group, the number of neuron activity and normal nuclei in MK-801 preconditioning group was significantly higher than that in the control group, but still lower than that in the control group. The difference was statistically significant (P <0.05). Conclusion The NR2B receptor mediates glutaraldehydeuria type I metabolism accumulation in striatal neurons.