锰诱导SH-SY5Y细胞线粒体自噬

Manganese induces mitophagy in SH-SY5Y cells

目的探讨锰能否诱导SH-SY5Y细胞发生线粒体自噬。方法以人神经母细胞瘤SH-SY5Y细胞为模型,不同剂量的Mn Cl2处理细胞24 h,通过MTT法检测Mn Cl2对细胞存活率的影响,流式细胞仪检测Mn Cl2对细胞线粒体膜电位的影响,以Mito-Tracker Green和Lyso-Tracker Red荧光探针分别标记线粒体和溶酶体,激光共聚焦显微镜观察细胞线粒体和溶酶体的共定位,以表征线粒体自噬。结果 Mn Cl2可剂量依赖性地降低SH-SY5Y细胞存活率,并降低细胞线粒体膜电位,Mn Cl2诱导细胞线粒体和溶酶体共定位。结论 Mn Cl2可诱导SH-SY5Y细胞发生线粒体自噬,线粒体自噬可能参与锰神经毒性机制。

Objective To explore whether manganese could cause mitophagy in human SH-SY5Y dopaminergic neuroblastoma cells as an in vitro model of dopaminergic neuron. Methods SH-SYSY cells were treated with 0, 0.25, O. 5 and 1.0 mmol/L manganese chloride （MnC12 ） for 24 h. MTT colorimetry test was used to detect the survival state of SH-SY5Y cells, the mitochondrial membrane potential （MMP） of SH-SY5Y cells was detected by flow cytometry （FCM）. Mitophagy was detected with laser scanning confocal microscopy to observe mitoehondria and lysosomes co-localization, which were respectively labeled by probes of Mito-Tracker Green and Lyso-Tracker Red. Results MnCI2 could dose-dependently suppress the viability of SH-SY5Y cells and result in the loss of the mitochondfial membrane potential. Mitochondria and lysosomes suhcellular co-localization was observed in the MnC12 treatment groups. Conclusions MnC12 could cause the mitophagy in SH-SY5Y cells and this may be involved in the manganese-induced neurotoxicity.