The Role Of Oxidative Stress In Acrolein Phthalate-induced DNA Damage In INS-1

Objective:Acrolein(propenal,acrylaldehyde),an ?,?-unsaturated aldehyde,which is a ubiquitous environmental contaminant to which humans are exposed through multiple pathways especially in the kitchen smoke-lampblack and cigarette smoking.The International Agency for Research on Cancer affirmed that there was insufficient proofs for the carcinogenicity in experimental cells and animals.But human-exposures can immediately exert via the dermal,inhalation and oral routes.In addition there is a increasing number of animal experiments evidence that acrolein is genotoxic,such as cardiovascular system,respiratory system,nervous system,reproductive system.This study observing the acrolein induced INS-1 cells the DNA the generation of ROS,the level of GSH and mitochondrial membrane in order to provide experimental and theoretical basis.Thus it may clarify the mechanisms of acrolein-induced on human health effects.Method: INS-1 cells were exposed to different doses of acrolein(0 ?12.5 ?25 ?50).Experiment applying single-cell gelelectrophoresis(SCGE)is examined to test DNA damage caused by acrolein and confirm the acrolein toxicity of DNA damage.To inspect the mechanism of DNA damage,with OPT,DCFH and rhodamine123 were determined acrolein on intracellular tri-peptide glutathione(GSH),reactive oxygen species(ROS),the effect on the potential of mitochondrial membrane.We discovered the vital role of oxidative associated destruction in DNA damage by PQQ intervention experiment.The whole results was analyzed by utilizing SPSS v17.0 statistical software.Results: In this study,the different doses of acrolein(0?M?12.5?M?25?M?50?M).caused a dose-dependent decrease in cell viability in INS-1 cells.When INS-1 cells exposed in acrolein after 1h,the comet experiments show double-stranded DNA rupture,compared with blank group under microscope,cell into a comet-like tail,with acrolein concentrations increasing,the long tail,tail DNA% and DNA% content increased markedly;the research declare that acrolein induced the increased levels of GSH level,the expression of ROS and mitochondrial membrane potential in INS-1 cells.After pretreatment with PQQ 1h,acrolein relieved INS-1 cells DNA damage,intracellular ROS and mitochondrial membrane potential have hugely declined,so it also can tremendously reduce intracellular reactive oxygen.Conclusion: Acrolein can lead to the DNA damage of INS-1 cells,illustrating its genetic toxicity.The exposure to various doses of acrolein lead to the content of intracellular ROS increased,the level of GSH reduced and.mitochondrial membrane potential.decreased.While antioxidant PQQ intervention trials have demonstrated that it can decrease the generation of intracellular reactive oxygen,can retard the level of DNA strand breaks and the expression of GSH generation.Meanwhile,it can make stability of mitochondrial membrane showed a trend of protection.All of these results evidence the effects of acrolein-induced DNA damage were related to oxidative stress.