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PCMT1/MST1通路在甲基丙二酸血症脑损伤中的作用
作者:吕书博  张展  赵德华  李晓乐  苏立  罗春伟  孟云  王丽雯 
单位:郑州大学第三附属医院 新生儿筛查中心, 河南 郑州 450000
关键词:甲基丙二酸 神经元 蛋白异天冬氨酸甲基转移酶1 哺乳动物不育系20样激酶1 凋亡 小鼠 
分类号:R-33;R361.3;R363.13
出版年·卷·期(页码):2019·38·第四期(667-671)
摘要:

目的:探讨PCMT1/MST1通路在甲基丙二酸血症脑损伤中的作用。方法:培养小鼠皮层神经元,用不同浓度甲基丙二酸诱导小鼠皮层神经元,观察细胞形态变化。采用四甲基偶氮唑蓝(MTT)法检测小鼠皮层神经元活力,Western blotting法检测各组(甲基丙二酸诱导组、CGP3466B治疗组和对照组)神经元蛋白异天冬氨酸甲基转移酶1(PCMT1)、磷酸化蛋白激酶(MST1)、裂解MST1(cl-MST1)、磷酸化MST1(p-MST1)、Caspase-3、B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)蛋白水平。结果:不同浓度甲基丙二酸诱导24 h显示,甲基丙二酸浓度≤8 mmol·L-1时神经元形态无明显变化;甲基丙二酸浓度为12 mmol·L-1时神经元胞内出现空泡,神经元突起缩短,胞体皱缩;甲基丙二酸浓度为16 mmol·L-1时神经元胞体脱落,突起几乎全部断裂。甲基丙二酸组神经元PCMT1、MST1蛋白相对表达量均低于对照组和CGP3466B治疗组(P<0.05),cl-MST1、p-MST1蛋白相对表达量均高于对照组和CGP3466B治疗组(P<0.05)。甲基丙二酸组Caspase-3、Bax蛋白相对表达量均高于对照组和CGP3466B治疗组(P<0.05),Bcl-2蛋白相对表达量均低于对照组和CGP3466B治疗组(P<0.05)。结论:甲基丙二酸可引起皮层神经元凋亡,这可能是通过PCMT1/MST1通路来实现的。

Objective: To observe the effect of the methylmalonic acid on mouse cortical neurons and cells protein isoaspartate methyltransferase 1(PCMT1)/mammalian sterile line 20-like kinase 1(MST1) pathway in brain injury caused by methylmalonic acid. Methods: The mouse cortical neurons were cultured. Methylmalonic acid with different concentrations induced mouse cortical neuronal cells and the changes of cell morphology were observed.MTT assay was used to detect the activity of cortical neurons in mice. The protein levels of PCMT1, MST1, cl-MST1, p-MST1, Caspase-3, B-cell lymphoma-2 (Bcl-2), and Bcl-2 related X protein (Bax) were detected by Western blotting. Results: After 24 hours of induction with different concentrations of methylmalonic acid, it was observed under microscope that the morphology of neurons did not change significantly when the concentration of methylmalonic acid ≤ 8 mmol·L-1, but vacuoles appeared in the neurons when the concentration of methylmalonic acid was 12 mmol·L-1, and the neurite processes were shortened. The cell body shrinked; When the concentration of methylmalonic acid was 16 mmol·L-1, the neuron body fell off and almost all the processes were broken, and methylmalonic acid significantly decreased the activity of neuron cells. The relative expression of PCMT1, MST1 protein in methylmalonic acid group was lower than those in control group and CGP3466B group (all P<0.05), and the expression of cl-MST1,p-MST1 protein was higher than those in the control group and the CGP3466B group (all P<0.05). The expression of Caspase-3 and Bax protein in methylmalonic acid group was higher than those in the control group and the CGP3466B group (all P<0.05), and the expression of Bcl-2 protein was lower than that in the control group and the CGP3466B group (P<0.05).Conclusion: Methylmalonic acid can induce apoptosis in cortical neurons. The mechanism may be that methylmalonic acid induces neuronal apoptosis through the PCMT1/MST1 pathway.

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