利美尼定促进突变SOD1蛋白质降解的作用研究

目的利美尼定对家族性肌萎缩侧索硬化症(ALS)相关突变蛋白质SOD1G93A的作用机制.方法用瞬时转染的方法在运动神经元样细胞系NSC-34中过表达WTSOD1,与家族型ALS相关的SOD1G93A突变蛋白,再给予自噬通路的特异性诱导剂和阻断剂,通过Western blot蛋白印迹法检测突变SOD1、自噬标记物的蛋白质表达水平.结果自噬诱导剂trehalose可以使SOD1G93A蛋白质表达水平明显减少.在自噬阻断剂3-甲基腺嘌呤(3-MA)的干预下,SOD1G93A蛋白质表达水平明显升高.l0uM利美尼定能够明显降低G93A SOD1的表达,但对WT SOD1的表达水平无明显影响.结论SOD1G93A主要经由自噬途径降解,利美尼定能够明显促进G93A SOD1的降解,但对WT SOD1的蛋白质表达水平无明显促进作用.

Study on the mechanism of rilmenidine promoting mutant SOD1 degradation

Objective To explore the mechanism of SOD1G93A degradation,a mutant protein associated with familial amyotrophic lateral sclerosis(ALS).Methods Transient transfection was used to overexpress WT S0D1 and the SOD1G93A protein associated with familial ALS in the motor neuron-like cell line NSC-34.Then,specific inducers and blockers of autophagy pathway were administered.Western blot immunoblotting was used to detect the protein expression levels of mutant S0D1 and autophagy markers.Results-The autophagy inducer trehalose can significantly reduce the protein expression level of SOD1G93A.Under the intervention of the autophagy blocker 3-MA,the protein expression level of SOD1G93A increased significantly.IOuM Rilmenidine can significantly reduce the expression of SOD1G93A,but has no obvious effect on the expression level of WT S0D1.Conclusion SOD1G93A is mainly degraded through autophagy pathway.Rilmenidine can significantly promote the degradation of G93A S0D1,but has no obvious effect on the degradation of WT S0D1.