ABSTRACT
OBJECTIVE: To investigate the effects of nicotinamide adenine dinucleoside phosphate oxidases 1(NOX1) and nuclear factor-kappa B(NF-κB) in tumor necrosis factor-α(TNF-α)-induced oxidative damage in A549 cells. METHODS: i) TNF-α was used to stimulated A549 cells at the concentrations of 0.0, 10.0, 25.0 and 50.0 μg/L. Cell inhibition rate in each group was tested by CCK-8 assay to select the appropriate concentration. ii) A549 cells in logarithmic growth phase were divided into blank control group, solvent control group, TNF-α group, diphenylene iodine(DPI) group and TNF-α+DPI group for NOX1 inhibitor experiment. Logarithmic growth phase A549 cells were divided into blank control group, TNF-α group, BAY11-7082 group and TNF-α+BAY11-7082 group for NF-κB inhibitor experiment. The relative expression of NOX1 and p65 protein in each group was detected by Western blot method. The relative expression of intracellular reactive oxygen species(ROS) were detected by flow cytometry. RESULTS: i) The inhibition rate of A549 cells increased with the increase of TNF-α dose(P<0.05), and 25.0 μg/L was selected as the stimulation dose of TNF-α in subsequent experiments. ii) The relative expression of NOX1, p65 protein and ROS in the TNF-α group was higher than that in the blank control group, solvent control group and DPI group, respectively(P<0.05). The above indexes in TNF-α+DPI group were lower than that in TNF-α group(P<0.05), but higher than that in DPI group(P<0.05). The relative expression of NOX1, p65 protein and ROS in the TNF-α group were higher than that in the blank control group and the BAY11-7082 group(P<0.05), while the above indicators in the TNF-α+BAY11-7082 group were lower than that in the TNF-α group(P<0.05). CONCLUSION: Inhibition of NOX1 or NF-κB can alleviate the oxidative damage induce by TNF-α in A549 cells.
ABSTRACT
Aim To detect the expression of hippocam-pus NGF gene in the mouse model of Parkinson 's dis-ease. Methods By intraperitoneal injection of 1-meth-yl-4-phenyl-1,2, 3, 6-tetrahydropyridine ( MPTP), a mice model of Parkinson ’ s disease was established. The success of PD model was identified by detecting the expression of mesencephalic tyrosine hydroxylase ( TH) with Western blot and immunofluorescence. The movement and cognitive ability of mice were evaluated by foot print analysis and step-through passive avoid-ance test. The expression of NGF gene in hippocampus of mice was detected with RT-PCR and in situ hybrid-ization. Results Compared with the control group, the levels of TH and NGF were reduced in the experi-mental group and the behavioral indexes were deflected significantly. Conclusion NGF may play an impor-tant role in the occurrence and development of the PD cognitive disorder.