Effects of METH on mitochondrial membrane potrntial, ultrastaucture and expression of Mfnl and Fisl protein in SH-SY5Y cells cultured in vitro / 中国药理学通报

ABSTRACT

Aim To evaluate the influence of METH on MMP, mitochondrial ultrastructure, and the expression levels of mitochondrial proteins, Mfnland Fisl, in human neuroblastoma SH-SY5Y cells in vitro. Methods A stable and feasible culture method of SH-SY5Y cells in vitro was established with different concentrations of METH(0. 0, 1. 0, 1. 5 and 2. 0 mmol L-1), and for various periods of exposure for 3, 6, 12, 24 h, the MMP of SH-SY5Y cells was stained by MMP assay kit (JC-1) , the mitochondrial ultrastructure of SH-SY5Y cells exposed to METH was observed by transmission electron microscope, and the expression levels of Mfnl and Fisl proteins were detected by Western blot. Results Compared with control group for various periods of exposure(3,6,12,24 h), the red/green fluorescence ratios of MMP and the expression levels of Mfn1 protein decreased significantly in METH groups (P<0. 05) , while the expression levels of Fisl pro-tein increased significantly (P <0.05). SH-SY5Y cells were treated with METH for 24 h prior to observation under transmission electron microscope ( TEM ). The mitochondria of SH-SY5Y cells in unprocessed group showed the oval, rodlike and double-layer membrane structure, along with clear normal mitochondrial cristae. However, the oval and rodlike structure of mitochondria in SH-SY5Y cells of METH treatment groups had been split into small ball structures. Moreover , mitochondrial autophagosome and autophagic iyso-some could also be found. Conclusions METH could induce a decrease in MMP, mitochondrial ultrastruc-tural changes, and changes in the expression levels of Mfnl and Fisl in SH-SY5Y cells, which might be associated with nerve cell damage caused by METH.