论文部分内容阅读
目的探讨一氧化氮合酶抑制剂(nitric oxide synthase inhibitor,NOS inhibitor)在N-甲基-D-天(门)冬氨酸(N-Methyl-D-Asparate,NMDA)诱导的兴奋性神经毒中的作用及其可能的作用机制。方法原代培养24 h内新生SD大鼠脑组织皮层神经元,利用免疫荧光方法鉴定细胞纯度。将培养的神经元随机分为对照组、NMDA染毒组和NMDA+NG-硝基-L-精氨酸甲酯(L-NAME)染毒组,每组24个平行。NMDA+L-NAME染毒组加入终浓度为100μmol/L的L-NAME染毒1 h后,在NMDA+L-NAME染毒组和NMDA染毒组分别加入终浓度为100μmol/L的NMDA染毒2 h。测定神经元细胞的活性、NO浓度、细胞凋亡情况及磷酸化p38丝裂原活化蛋白激酶(p-p38MAPK)的表达。结果免疫荧光显示90%细胞为兔抗大鼠神经元特异性烯醇化酶(neuronal specific enolase,NSE)阳性细胞;与对照组相比,NMDA染毒组与NMDA+L-NAME染毒组大鼠脑组织神经元中NO的浓度、细胞凋亡率及p-p38MAPK的表达明显升高,细胞活性明显下降,差异均有统计学意义(P<0.01);而与NMDA染毒组相比,NMDA+L-NAME染毒组大鼠脑组织神经元中NO的浓度、细胞凋亡率及p-p38MAPK的表达明显降低,细胞活性明显上升,差异有统计学意义(P<0.01)。结论 NMDA诱导的神经细胞凋亡部分依赖NO毒性作用,抑制NO的毒性作用可部分抑制神经细胞凋亡。
Objective To investigate the effects of NOS inhibitor on excitatory neurotoxicity induced by N-methyl-D-asparate (NMDA) In the role and possible mechanism of action. Methods Primary cortex neurons of neonatal SD rats were cultured within 24 h. The purity of cells was identified by immunofluorescence. The cultured neurons were randomly divided into control group, NMDA-treated group and NMDA-N-nitro-L-arginine methyl ester group (L-NAME). NMDA + L-NAME and L-NAME at a final concentration of 100 μmol / L for 1 h, NMDA + NMDA + NMDA at a final concentration of 100 μmol / L Poison 2 h. The activity of neurons, the concentration of NO, the apoptosis and the expression of phospho-p38 mitogen-activated protein kinase (p-p38MAPK) were measured. Results Immunofluorescence showed that 90% of the cells were anti-rat neuron-specific enolase (NSE) positive cells. Compared with the control group, NMDA + L-NAME treatment group Compared with NMDA group, the concentration of NO, the apoptosis rate and the expression of p-p38MAPK in brain tissue neurons increased significantly, and the cell activity decreased significantly (P <0.01) The concentration of NO, the apoptosis rate and the expression of p-p38MAPK in neurons of + L-NAME exposure group were significantly decreased, and the cell activity was significantly increased (P <0.01). Conclusion NMDA-induced neuronal apoptosis partly depends on NO toxicity and inhibition of NO toxicity partly inhibits neuronal apoptosis.