锰中毒对SH-SY5Y细胞线粒体自噬作用的影响

Effect of Manganese Poisoning on Mitophagy in SH-SY5Y Cells

[目的]探讨锰中毒对SH-SY5Y细胞线粒体自噬作用的影响。[方法]取对数生长期的人神经母细胞瘤SH-SY5Y细胞,予MnCl2处理24 h,采用酶联免疫吸附试验(ELISA)检测细胞中多巴胺(DA)的分泌量,蛋白免疫印迹法检测自噬标记蛋白LC3、线粒体分裂蛋白Drp1、线粒体融合蛋白Mfn1、Mfn2的表达。[结果]与正常对照组相比,MnCl2组多巴胺的分泌量显著减少(P<0.05),LC3-Ⅱ表达增加(P<0.05),Drp1表达降低(P<0.05),Mfn1及Mfn2表达增加(P<0.05)。[结论]锰中毒可显著降低SH-SY5Y细胞中多巴胺的分泌量,抑制线粒体的分裂并促进线粒体融合,导致线粒体自噬增多,这可能是锰中毒性帕金森综合征的病理机制之一。

[Objective]To investigate the effects of manganese poisoning on mitophagy in SH-SY5 Y cells.[Methods]Human neuroblastoma SH-SY5 Y cells in logarithmic growth phase were treated with MnCl2 for 24 h.Enzyme-linked immunosorbent assay(ELISA)was used to detect the secretion of dopamine(DA)in cells.Western blotting was used to detect the expression of autophagy marker protein LC3,mitochondrial protein Drp1,mitochondrial fusion proteins Mfn1,Mfn2.[Result]Compared with the normal control group,the secretion of dopamine in the MnCl2 group was significantly decreased(P<0.05),the expression of LC3-Ⅱwas increased(P<0.05),the expression of Drp1 was decreased(P<0.05),and the expression of Mfn1 and Mfn2 was increased(P<0.05).[Conclusion]Manganese poisoning can significantly reduce the secretion of dopamine in SH-SY5 Y cells,inhibit mitochondrial division,and promote mitochondrial fusion,leading to increased mitochondrial autophagy,which may be one of the pathological mechanisms of manganese-induced Parkinson’s syndrome.