Mitochondrial dysfunction enhances susceptibility to oxidative stress by down-regulation of thioredoxin in human neuroblastoma cells.
Increasing evidence suggests that Alzheimer's disease is associated with mitochondrial dysfunction and oxidative damage. To develop a cellular model of Alzheimer's disease, we investigated the effects of thioredoxin (Trx) expression in the response to mitochondrial dysfunction-enhanced oxidative stress in the SH-SY5Y human neuroblastoma cells. Treatment of SH-SY5Y cells with 15 mM of NaN3, an inhibitor of cytochrome c oxidase (complex IV), led to alteration of mitochondrial membrane potential but no significant changes in cell viability. Therefore, cells were first treated with 15 mM NaN3 to induce mitochondrial dysfunction, then, exposed to different concentrations of H2O2. Cell susceptibility was assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay and morphological observation. Expressions of Trx mRNA and protein were determined by RT-PCR; and Western-blot analysis, respectively. It was found that the SH-SY5Y cells with mitochondrial impairment had lower levels of Trx mRNA and protein, and were significantly more vulnerable than the normal cells after exposure to H2O2 while no significant changes of Trx mRNA and protein in SH-SY5Y cells exposed to H2O2 but without mitochondrial complex IV inhibition. These results, together with our previous study in primary cultured neurons, demonstrated that the increased susceptibility to oxidative stress is induced at least in part by the down-regulation of Trx in SH-SY5Y human neuroblastoma cells with mitochondrial impairment and also suggest the mitochondrial dysfunction-enhanced oxidative stress could be used as a cellular model to study the mechanisms of Alzheimer's disease and agents for prevention and treatment.
Ding
Hongqun
H
School of Pharmacy, Jiangsu University, Zhenjiang 212013, PR China.
Gao
Jing
J
Zhu
Zengrong
Z
Xiong
Yuyun
Y
Liu
Jiankang
J
D001483:Base Sequence; D015153:Blotting, Western; D045744:Cell Line, Tumor; D017931:DNA Primers; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008564:Membrane Potentials; D008928:Mitochondria / Q000502:physiology*; D009447:Neuroblastoma / Q000473:pathology*; D009584:Nitrogen / Q000494:pharmacology; D018384:Oxidative Stress; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013879:Thioredoxins / Q000378:metabolism*
D017931:DNA Primers; C003320:sodium nitride; D013879:Thioredoxins; D006861:Hydrogen Peroxide; D009584:Nitrogen