运动神经元病相关蛋白p150glued在自噬通路中的功能

The role of motor neuron disease associated protein p150glued in autophagy pathway

目的探讨运动神经元病相关蛋白p150glued在自噬通路中的作用和功能。方法以1.5个月龄C57BL/6J野生型小鼠和HEK293细胞为材料,利用转染和蛋白印迹(Western blot)法检测p150glued缺失对于Dynactin亚基表达水平以及自噬-溶酶体蛋白降解通路中关键蛋白的影响。结果p150glued在小鼠神经系统中表达广泛,且与脊髓相比,坐骨神经中其含量显著下降(P<0.05)。p150glued蛋白缺失引起动力蛋白激活蛋白Dynactin亚基DCTN4、DCTN5、p50和Arp1a蛋白表达水平显著下降(P<0.05)。p150glued蛋白含量下调使自噬-溶酶体蛋白降解通路中LC3A/B-II、p62和LAMP1蛋白表达水平显著提高(P<0.05),而ATG3、ATG7和LC3A/B-I蛋白含量未发生明显变化(P>0.05)。结论运动神经元病相关蛋白p150glued含量减少可通过引起Dynactin亚基表达水平下降,使自噬小体和溶酶体运输障碍,从而导致自噬-溶酶体蛋白降解通路异常。

Objective To investigate the role of p150 glued in autophagy pathway.Methods The expression of p150 glued,Dynactin subunits and autophagy-lysosome pathway key proteins of 1.5 months wildtype C57 BL/6 J mouse and HEK293 cells were examined by siRNA transfection and western blotting.Results p150 glued extensively expression in the nervous system of mice,and compared with the spinal cord,the content of p150 glued in sciatic nerve was relatively low(P<0.05).Knockdown of p150 glued reduced the level of DCTN4,DCTN5,p50 and Arp1 a(P<0.05),and improved the level of LC3 A/B-II,p62,LAMP1(P<0.05).Conclusion The loss of p150 glued can reduce the level of Dynactin subunits,resulting in the dysfunction of the autophagosomes and lysosomes transport,thus lead to the abnormal autophagy-lysosome protein degradation pathway.