NADPH氧化酶介导吉非替尼诱导的小鼠肝损伤OA

AIM:To investigate the role and molecular mechanism of nicotinamide adenine dinucleotide phos-phate(NADPH)oxidase in gefitinib(GEF)-induced liver injury of mice.METHODS:Sixty male BALB/c mice were randomly divided into normal control(NC)group,low,medium and high dose(50,100 and 200 mg·kg-1·d-1)GEF groups,with 15 mice in each group.All mice received oral gavage once daily for 2 weeks.Liver index and spleen index were calculated.Histopathological changes in the liver were observed by HE staining,and reactive oxygen species(ROS)production was detected using dihydroethidium staining.Levels of serum aspartate aminotransferase(AST)and alanine aminotransferase(ALT),as well as hepatic malondialdehyde(MDA)and glutathione(GSH)contents and superoxide dis-mutase(SOD)activity,were detected using commercial kits.Western blot was performed to investigate the protein expres-sion levels of NADPH oxidase catalytic subunits NOX2 and NOX4,inhibitor of nuclear factor κB kinase subunit α(IKKα),phosphorylated nuclear factor-κB p65(p-NF-κB p65)and NF-κB p65 in the liver of mice.Serum levels of tu-mor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were measured using ELISA.The above oxidative stress mark-ers,inflammatory cytokine levels,and the protein expressions of NOX2,NOX4,IKKα,p-NF-κB p65 and NF-κB p65 were detected in AML12 cells.N-acetylcysteine(NAC)intervention was performed to investigate the relationship between ROS and the NF-κB pathway.RESULTS:Compared with the NC group,all GEF-treated mice exhibited decreased body weight(P<0.01),increased liver index and spleen index,as well as elevated serum AST and ALT levels(P<0.05),and histopathological injuries including vacuolization,necrosis and inflammation in the liver.Furthermore,ROS production and MDA levels in liver tissue and the culture supernatant of AML12 cells were significantly increased(P<0.01),while SOD activity and GSH levels were decreased in GEF-treated groups(P<0.05).GEF also upregulated the protein expres-sions of NOX2,NOX4,IKKα and phosphorylation of NF-κB p65 in liver tissue of mice and AML12 cells(P<0.05).Meanwhile,TNF-α and IL-6 levels were increased in the serum of mice and in the culture supernatant of AML12 cells(P<0.05).NAC intervention significantly reversed GEF-induced ROS production and phosphorylation of NF-κB p65 in AML12 cells.CONCLUSION:GEF induces oxidative stress and inflammatory responses in the mouse liver,leading to liver injury.This effect may be mediated by upregulation of NOX2 and NOX4 and subsequent activation of the IKKα/NF-κB signaling pathway.