Influence of Hydrogen Peroxide on Mitochondrial Membrane Potential and Superoxide Production in FRTL Cell / 天津医药

ABSTRACT

Objective:To investigate the effects of exogenous hydrogen peroxide(H_2O_2)on mitochondrial superoxide production and mitochondrial membrane potential changes(△ψ)in fisher rat thyroid cell line(FRTL).Methods:Following 1 mmol/L H_2O_2 exposure in FRTL cells for 10 min,30 min and 24 h,mitochondrial superoxide production was measured by living cell imaging and flow cytometry using MitoSOX.The mitochondrial membrane potential was assayed by spectrofluorimeter and fluorescent microscopy using rhodamine 123(rh123).The cell viability was detected by MTT colorimetric method.Morphological changes were observed by invert microscope.Apoptosis assay was performed by acridine orange staining.Results:Quantitative measurements of the mean intensities of MitoSOX demonstrated significant increase with 1 mmol/L H_2O_2 following 10 min,30 min and 24 h treatment in FRTL cells compared with that of control.Fluorescence intensity of rh123 and optical density of MTr were significantly decreased in FRTL ceils with 1 mmol/L H_2O_2 following 30 min and 24 h treatment(P ＜ 0.01).Under light microscope and fluorescence microscope the characteristic morphological features of programmed cell death,pickuosis,karyorrhexis,and cell shrinkage were observed after acridine orange staining.Conclusion:Acute and chronic exogenous H_2O_2 exposure cause oxide stress in FRTL cells,which result in the increase of mitochondrial superoxide production,△ψdecline,cell necrosis and apoptosis.