Journal of Jilin University(Medicine Edition) ›› 2020, Vol. 46 ›› Issue (6): 1117-1123.doi: 10.13481/j.1671-587x.20200602

• Research in basic medicine • Previous Articles     Next Articles

Protective effect of rutin on oxidative stress injury of HepG2 cells and its mechanism

Fangduo JIN,Tian ZHANG,Zhao ZHANG,Xuezhe YIN,Jishu QUAN()   

  1. Department of Biochemistry and Molecular Biology,School of Basic Medical Sciences,Yanbian University,Yanji 133002,China
  • Received:2020-05-12 Online:2020-11-28 Published:2022-08-24
  • Contact: Jishu QUAN E-mail:quanjs1@sina.com

Abstract: Objective

To explore the protective effect of rutin on the oxidative stress injury of liver cancer HepG2 cells, and to clarify its mechanism.

Methods

The liver cancer HepG2 cells in the logarithmic growth phase were divided into control group, tert-butyl hydroperoxide (TBHP) group (400 μmol·L-1 TBHP), 0.25, 0.50,and 1.00 μmol·L-1 rutin groups (0.25, 0.50, 1.00 μmol·L-1 rutin+400 μmol·L-1 TBHP). The vitalities of liver cancer HepG2 cells in various groups and the survival rates of liver cancer HepG2 cells in various groups were detected by MTT assay;the levels of malondialdehyde (MDA) and glutathione (GSH) and activities of superoxide dismutase (SOD) in the liver cancer HepG2 cells in various groups were assessed by colorimetric method; the levels of reactive oxidative species (ROS) in the liver cancer HepG2 cells in various groups were quantified with DCFH-DA immnofluorescence staining. The expression levels of PI3K, p-PI3K, Akt, p-Akt, NF-κB, p-NF-κB,and p53 proteins in the liver cancer HepG2 cells in various groups were mesured by Western blotting method.

Results

The MTT results showed that when the concentrations of rutin were less than or equal to 1.00 μmol·L-1, the vitality of liver cancer HepG2 cells was higher than 90%, and the rutin had no toxic effect; when the concentration of rutin was higher than 1.00 μmol·L-1, the vitality of the liver cancer HepG2 cells was lower than 90%,and the rutin had toxic effect. Compared with control group, the survival rate of the HepG2 cells in TBHP group was significantly decreased (P<0.01); compared with TBHP group, the survival rates of the HepG2 cells in different concertrations of rutin groups were significantly increased (P<0.01). Compared with control group, the level of MDA in the HepG2 cells in TBHP group was significantly decreased (P<0.01), the level of GSH and the activity of SOD were significantly increased (P<0.01); compared with TBHP group, the levels of MDA in the HepG2 cells in different concentrations of rutin groups were significantly increased(P<0.01), the levels of GSH and the activities of SOD in the HepG2 cells in different concertrations of rutin groups were significantly decreased (P<0.01). The results of DCFH-DA immunofluorescence staining showed that the ROS level in the HepG2 cells in TBHP group was higher than that in control group; compared with TBHP group, the ROS levels in the HepG2 cells in different concentrations of rutin groups were gradually decreased(P<0.05). The Western blotting results showed that compared with control group, the expression levels of PI3K, p-PI3K, Akt, and p-Akt proteins in the HepG2 cells in TBHP group were decreased (P<0.05), and the expressions levels of NF-κB, p-NF-κB,and p53 proteins were increased (P<0.05); compared with TBHP group, the expressions levels of PI3K, p-PI3K, Akt, and p-Akt proteins in the HepG2 cells in different concentrations of rutin groups were increased (P<0.05), and the expressions levels of NF-κB, p-NF-κB, and p53 proteins were decreased (P<0.05).

Conclusion

Rutin has the protective effect on TBHP-induced HepG2 cell damage, and its mechanism may be related to the regulation of PI3K/Akt and NF-κB pathways.

Key words: rutin, tert-butyl hydroperoxide, oxidative stress, hepatocyte injury, hepatocyte protection

CLC Number: 

  • R282.7