【摘要】 目的:观察甲基丙二酸对小鼠皮层神经元凋亡及细胞中c-jun氨基末端激酶（JNK）、磷酸化c-jun氨基末端激酶（p-JNK）、c-fos、c-jun蛋白水平的影响,探讨甲基丙二酸血症脑损伤的机制。方法:培养小鼠皮层神经元,甲基丙二酸诱导小鼠皮层神经元,采用流式细胞术检测小鼠皮层神经元凋亡情况。将神经元分为对照组（加入正常培养基）、甲基丙二酸组(加入甲基丙二酸浓度为15 mmol·L^-1的培养基)和JNK抑制剂组(加入甲基丙二酸浓度为15 mmol·L^-1和SP600125浓度为40μmol·L^-1的培养基),采用蛋白质印迹法检测各组神经元JNK、p-JNK、c-fos、c-jun、B淋巴细胞瘤-2基因（Bcl-2）、Bcl-2相关X蛋白（Bax）、Caspase 3、cleaved-caspase 3蛋白表达水平。结果:与对照组比较,甲基丙二酸浓度为5、10 mmol·L^-1时凋亡神经元比例变化不明显（P>0.05）,甲基丙二酸浓度为15 mmol·L^-1时凋亡神经元比例明显增加（P<0.05）。甲基丙二酸组神经元p-JNK、c-fos、c-jun蛋白相对表达量高于对照组（P<0.05）,JNK蛋白相对表达量和对照组比较差异无统计学意义（P>0.05）。甲基丙二酸组Bax、Caspase 3、cleaved-caspase 3蛋白相对表达量高于对照组（P<0.05）,Bcl-2蛋白相对表达量低于对照组（P<0.05）;JNK抑制剂组Bax、Caspase 3、cleaved-caspase 3蛋白相对表达量低于甲基丙二酸组（P<0.05）,Bcl-2蛋白相对表达量高于甲基丙二酸组（P<0.05）。结论:甲基丙二酸可引起小鼠皮层神经元损伤,其机制可能为甲基丙二酸通过JNK/MAPK通路诱导小鼠皮层神经元凋亡。更多还原

【Abstract】 Objective: To observe the effect of methylmalonic acidemia on the apoptosis of cortical neurons and the c-jun N-terminal kinase（JNK）, phosphorylation c-jun N-terminal kinase（p-JNK）, c-fos, c-jun protein levels in cortical neuronal, and to explore the mechanism of brain damage induced by methylmalonic acidemia. Methods:The mouse cortical neuronal cells were cultured. The mouse cortical neuronal cells were induced by methylmalonic acidemia. Flow cytometry was used to detect the apoptosis of mouse cortical neurons. Neuron cells were divided into control group（added to normal medium）, methylmalonic acidemia group(added to medium with methylmalonic acidemia concentration of 15 mmol·L^-1) and JNK inhibitor group(added to medium with methylmalonic acidemia concentration of 15 mmol·L^-1 and SP600125 concentration of 40 μmol·L^-1).Western blotting was used to detect the protein levels of JNK, p-JNK, c-fos, c-jun, B-lymphoma-2（Bcl-2）, Bcl-2 related X protein（Bax）, Caspase 3 and cleaved-caspase 3 in neurons. Results:Compared with the control group, when the concentration of methylmalonic acidemia were 5 mmol·L^-1 and 10 mmol·L^-1, the change of the proportion of apoptotic neurons was not obvious（P>0.05）, when the concentration of methylmalonic acidemia was 15 mmol·L^-1, the proportion of apoptotic neurons increased significantly（P<0.05）. The relative expressions of p-JNK, c-fos and c-jun proteins in the neurons of methylmalonic acidemia group were higher than those in the control group（P<0.05）. There was no significant difference in the relative expression of JNK protein between the methylmalonic acidemia group and the control group（P>0.05）. The relative expressions of Bax,Caspase 3 and cleaved-caspase 3 protein in the methylmalonic acidemia group were higher than those in the control group（P<0.05）. The relative expression of Bcl-2 protein in the methylmalonic acidemia group was lower than that in the control group（P<0.05）. The relative expressions of Bax, Caspase 3 and cleaved-caspase 3 protein in the JNK inhibitor group were lower than those in the methylmalonic acidemia group（P<0.05）, and the relative expression of Bcl-2 protein in the JNK inhibitor group was higher than that in the methylmalonic acidemia group（P<0.05）. Conclusion: Methylmalonic acidemia can induce cell damage of mouse cortical neurons. The mechanism may be that methylmalonic acidemia can induce apoptosis of mouse cortical neurons by the JNK/MAPK pathway.更多还原