The Role Of Mitochondrial Fusion And Fission Abnormality In Fluoride-induced Mitochondrial Injury In SH-SY5Y Cells

Part?The influence of NaF on mitochondrial functions and mitochondrial fusion/fission related protein expression in SH-SY5YcellsObjective:Long-term intake of excessive fluoride is neurotoxic,but its mechanism has not been elucidated.This research adopted different concentratons of sodium fluoride?NaF?infecting human neuroblastoma SH-SY5Y cells,to observe the influence of NaF on mitochondrial functions and mitochondrial fusion/fission related protein expression,to further clarify fluorine neurotoxic mechanism with relevant theoretical basis.Methods:SH-SY5Y cells were exposed to different concentration of NaF?20?40?60 mg/L?.After the cells were stained respectively by mitochondrial superoxide indicator MitoSOX^TM Red and mitochondrial membrane potential probe JC-10,the mitochondrial ROS?MitoROS?and mitochondrial membrane potential???m?were detected by flow cytometry.Meanwhile,western blot was used to measure mitochondrial fusion-related protein Mfn1?Mfn2 and fission-related protein Drp1?Fis1expression levels.Results:Along with the increase of NaF concentration,the MitoROS generation increased and the??m decreased.Compared with Control group,MitoROS in 60 mg/L NaF group increased significantly?P<0.05?.The??m in 20?40?60 mg/L NaF groups all decreased evidently?P<0.05?.The incremental expression levels of mitochondrial fusion-related protein Mfn1?Mfn2 were in accordence with the increase of NaF concentration and there was evident stastistical significance in 40?60mg/L NaF groups?P<0.05?.The fission-related protein Drp1?Fis1 expression levels in60 mg/L NaF group decreased significantly contracted with Control groups?P<0.05?.Conclusions:NaF could damage mitochondria.It could promote fusion and inhibit fission,resulting in the abnormal mitochondrial fusion and fisSion process.Part?The influence of mitochondrial fusion/fission on mitochondrial functional injury induced by NaF in SH-SY5Y cellsObjective:To investigate the influence of Mdivi-1?the inhibitor of mitochondrial fission?and NaF on mitochondrial fusion/fisSion protein expression levels and function changes in SH-SY5Y cells.Methods:SH-SY5Y cells were divided into four groups:Coutrol group,NaF-infected group?60 mg/L NaF?,Mdivi-1-treated group?10?M Mdivi-1?,Mdivi-1+NaF group?NaF infected cells after Mdivi-1 pretreated cells for30 min?.The protein expression levels of mitochondrial fusion-related protein Mfn1?Mfn2 and fission-related protein Drp1?Fis1 were detected by western blot method.Flow cytometry was used to detect MitoROS and??m after SH-SY5Y cells were stained respectively by MitoSOX^TM Red and JC-10.Results:Compared with the NaF-infected group,the expression levels of mitochondrial fusion-related protein Mfn1?Mfn2 in Mdivi-1+NaF group enhanced further?P<0.05?,fission-related protein Drp1and Fis1 declined further and there was a statistically significant decline of Fis1?P<0.05?.What's more,MitoROS increased further clearly?P<0.05?,??m decreased further?P<0.05?.Conclusions:Inbhibition of mitochondrial fission can improve the mitochondrial fusion obviously,indicating NaF can result in SH-SY5Y cells mitochondrial damage through influencing the mitochondrial fusion and fission process.