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| Study on the repair mechanism of Yiguanjian Decoction in DNA oxidative damage of L02 cells induced by H2O2 |
| WANG Amei ZHANG Rui LIU Wenlan |
| School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China |
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Abstract Objective To investigate the repair mechanism of Yiguanjian Decoction in hydrogen peroxide (H2O2)-induced DNA oxidative damage of human normal liver cells (L02). Methods L02 cells were randomly divided into 6 groups, including normal control group, model control group, low dose Yiguanjian Decoction group, middle dose Yiguanjian Decoction group, high dose Yiguanjian Decoction group and positive control group, five mice in each group. The normal control group and the model control group were cultured with medium containing 20% normal rat serum, simultaneously, low dose Yiguanjian Decoction group, middle dose Yiguanjian Decoction group, high dose Yiguanjian Decoction group were respectively cultured with medium containing 20% low, middle, high dose Yiguanjian Decoction of serum, and positive control group was cultured with 20% normal rat serum containing 0.1 mmol/L Vc. After 24 hours, except the normal control group, the cells of other groups were treated with 0.6 mmol/L H2O2 for 1 hour. The cell viability was detected by CCK-8; the content of 8-hydroxydeoxyguanosine (8-OHDG) was determined by enzyme linked immunosorbent assay (ELISA); the reactive oxygen species (ROS) contents and cell cycle were tested by flow cytometry; the mRNA levels of ATM, CHK2, Cdc25A and Cdc25C were assessed by revers transcriptation-polymerase chain reaction (RT-PCR). Results L02 cell viability was decreased with increasing H2O2 concentration and prolonged action time. Compared with the normal control group, cell viability of the model control group was decreased (P ＜ 0.05); 8-OHDG contents were increased (P ＜ 0.05); the mean fluorescence intensity of ROS was increased significantly (P ＜ 0.05); the cell proportion of G1 phase was decreased, while the cell proportion of S phase and G2/M phase were increased (P ＜ 0.05); the mRNA levels of ATM and CHK2 were up-regulated, and Cdc25A and Cdc25C were down-regulated (P ＜ 0.05). Compared with the model control group, the cell viability of the low dose Yiguanjian Decoction group, middle dose Yiguanjian Decoction group, high dose Yiguanjian Decoction group and positive control group were increased (P ＜ 0.05); the 8-OHDG content were decreased (P ＜ 0.05); the mean fluorescence intensity of ROS were decreased (P ＜ 0.05); the cell proportion of G1 phase were increased, the cell proportion of S phase and G2/M phase were decreased (P ＜ 0.05); the mRNA levels of ATM and CHK2 were down-regulated, while the expression of Cdc25A and Cdc25C mRNA were up-regulated (P ＜ 0.05). Conclusion Yiguanjian Decoction could repair the DNA oxidative damage of L02 cells by inhibiting the cell cycle through the ATM/CHK2/Cdc25A/Cdc25C pathway.
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