Protective Effect Of Rutin On TBHP-inducded Oxidative Injury Of Liver Cells

Objective:To study the protective effect and mechanism of rutin on oxidative injury induced by tert-butyl hydroperoxide(TBHP)in HepG2 cells.Methods:The oxidative injury model was established by treating HepG2 cells with TBHP.HepG2 cells were randomly divided into the control group,TBHP group,and low,medium and high-dose rutin groups.Cell survival rate of the HepG2 cells were determined by MTT assay.The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and lactate dehydrogenase(LDH)were detected by coloerimetric method.And the reduced glutathione(GSH),malondialdehyde(MDA)and superoxide dismutase(SOD)were measured by spectrophotometric method.The level of reactive oxygen species(ROS)was observed by DCFH-DA fluorescent staining.The cell apoptosis was evaluated by the TdT mediated dUTP nick end labeling(TUNEL)assay.The protein expressions of PI3K,p-P13K,Akt,p-Akt,NF-κB,p-NF-κB,IKKβ,p53 and PARP were determined by western blot method.Results:After pretreatment with rutin for 24 hours,compared with the TBHP group,rutin significantly increased the survival rates of HepG2 cells(P<0.01),and decreased the activities of AST,ALT and LDH in the cell culture medium(P<0.05)Rutin also reduced the content of MDA(P<0.05),increased intracelluar SOD activity and GSH content(P<0.05),reduced the ROS level(P<0.05),and suppressed cell apoptosis of HepG2 cells.Western blot results showed that rutin up-regulated the protein expressions of PI3K,p-PI3K,Akt,p-Akt(P<0.05),down-regulated the protein expressions of NF-κB,p-NF-κB,IKKβ(P<0.05),up-regulated the protein expressions of PARP and down-regulated the protein expressions of p53(P<0.05).Conclusion:Rutin has the protective effect on the oxidative damage induced by TBHP in HepG2 cells,maybe via PI3K/Akt and NF-κB signaling pathways.