论文部分内容阅读
Objective To explore the production and cytotoxicity of the reactive oxygen species(ROS)induced by diallyl trisulfid(DATS)in HL-60 cells.Methods HL-60 cells were either treated with various doses of DATS alone,or DATS combination with Apocynin,a specific NADPH oxidase inhibitor,or with antioxidant N-acetyl-L-cysteine(NAC)for 0,1,3,6,12 and 24 hours,respectively.The intracellular ROS level was measured by flow cytometry.The activity of NADPH oxidase was evaluated by NBT reduction experiment.The content of both malondialdehyde(MDA)and the protein carbonyl was analyzed by spectrophotometer.Results The results from flow cytometry indicated that DATS significantly increased the intracellular ROS level in HL-60 cells(P<0.05),which is a dose-and time-dependent.The fluorescence intensities of ROS reached at maximuam when HL-60 cells were incubated with 150 μmol·L-1 DATS for 3 hours.The NBT reduction experiment showed that DATS activated NADPH oxidase which had highest activity when cell were exposed to 150 μmol·L-1 DATS for 3 hours.Results DATS induced MDA and protein carbonyl production in HL-60 cells.Furrthermore,both MDA and protein carbonyl in the cells exposed to 150 μmol·L-1 DATS for 3 hours reached the highest level.Apocynin and NAC could attenuate the production of MDA and protein carbonyl,which suggested that ROS induced by DATS was involved in the toxicity to cells.Conclusions DATS induce ROS production through activating NADPH oxidase in HL-60 cells.ROS induced by DATS increase the oxidation of the membrane lipid and the protein of HL-60 cell.
Objective To explore the production and cytotoxicity of reactive oxygen species (ROS) induced by diallyl trisulfid (DATS) in HL-60 cells. Methods HL-60 cells were either treated with various doses of DATS alone, or DATS combination with Apocynin, a specific NADPH oxidase inhibitor, or with antioxidant N-acetyl-L-cysteine (NAC) for 0,1,3,6,12 and 24 hours, respectively.The intracellular ROS level was measured by flow cytometry. The activity of NADPH oxidase was evaluated by NBT reduction experiment. The content of both malondialdehyde (MDA) and the protein carbonyl was analyzed by spectrophotometer. Results of the results from flow cytometry indicated that DATS significantly increased the intracellular ROS level in HL-60 cells (P <0.05), which is a dose-and time-dependent. The fluorescence intensities of ROS reached at maximuam when HL-60 cells were incubated with 150 μmol·L-1 DATS for 3 hours. NBT reduction experiment showed that DATS activated NADPH oxidase which had highest activity when cell were exposed to 150 μmol·L-1 DATS for 3 hours. Results DATS induced MDA and protein carbonyl production in HL-60 cells. Both the MDA and protein carbonyl in the cells exposed to 150 μmol·L-1 DATS for 3 hours reached the highest level. Apocynin and NAC could attenuate the production of MDA and protein carbonyl, which suggested that ROS induced by DATS was involved in the toxicity to cells. Conclusions DATS induce ROS production through activating NADPH oxidase in HL-60 cells. ROS induced by DATS increase the oxidation of the membrane lipid and the protein of HL-60 cell.