Effect of apigenin on H2O2-induced oxidative injury in human hepatocytes L02

Objective To investigate the protective effect of apigenin against H2O2-induced injury in human hepatocytes(L02).Methods L02 cells were treated with H2O2 to establish a model of oxidative injury. CCK-8 assay was used to measure cell viability; DCFH-DA was used to measure the production of reactive oxygen species(ROS) in cells; test kits were used to measure the activities of lactate de-hydrogenase(LDH), malondialdehyde(MDA), and superoxide dismutase(SOD) in cell supernatant; Hoechst staining was performed toobserve cell apoptosis, and a test kit was used to observe the activity of caspase-3. A one-way analysis of variance was used for compari-son of continuous data between multiple groups, and the LSD-ttest was used for further comparison between two groups.Results Apige-nin at a concentration of ≥20 μmol/L significantly inhibited the proliferation of L02 cells(P< 0. 01). Compared with the cells in the blankcontrol group, the cells treated with H2O2 at a concentration of ≥500 μmol/L had a significant reduction in cell viability(P< 0. 001), andtherefore, 500 μmol/L was determined as the optimal concentration for modeling. There was a significant difference in cell viability betweenthe model group and the blank control group(P< 0. 01), and compared with the model group, the 5 and 10 μmol/L apigenin groups had asignificant increase in cell viability(P< 0. 01). The cells in the blank control group had good morphology, while those in the model groupwere contracted and round-shaped and had marked rupture and deformity, and compared with the model group, the 5 μmol/L apigenin group showed significant improvement with a reduction in ruptured and round-shaped cells. There was a significant difference in fluores-cence intensity between the blank control group, the model group, and the 5 μmol/L apigenin group(1. 00 ± 0. 26 vs 32. 94 ± 1. 29 vs13. 49 ± 1. 23,F= 1. 10,P< 0. 001), and the model group had a significantly higher fluorescence intensity than the blank control group(P< 0. 001). Compared with the model group, the 5 μmol/L apigenin group had a significant reduction in H2O2-induced ROS(P<0. 001). Compared with the control group, the model group had significant increases in the levels of LDH and MDA and a significant reduc-tion in the level of SOD(F= 3. 21, 2. 03, and 3. 32, allP< 0. 05), and compared with the model group, the 5 μmol/L apigenin group hadsignificant reductions in the levels of LDH and MDA and a significant increase in the level of SOD(allP< 0. 05). There was a significantdifference in cell apoptosis rate between the blank control group, the model group, and the 5 μmol/L apigenin group(7. 54% ± 0. 52% vs39. 77% ± 3. 44% vs 14. 40% ± 0. 79%,F =9.439,P< 0. 01], and the model group had a significantly higher cell apoptosis rate than thecontrol group(P< 0. 01); the cells treated with apigenin had a significantly lower apoptosis rate than those in the model group(P< 0. 01).There was a significant difference in the activity of caspase-3 between the blank control group, the model group, and the 5 μmol/L apigeningroup(4. 38 ±0. 59 U/mg vs 16. 44 ±1. 13 U/mg vs 10. 60 ± 1. 04 U/mg,F =1.17,P< 0. 05), and the model group had a significantlyhigher activity of caspase-3 than the blank control group(P< 0. 05); compared with the cells in the model group, the cells treated withapigenin had a significant reduction in the activity of caspase-3(P< 0. 05).Conclusion Apigenin may exert a protective effect againstH2O2-induced injury in L02 cells by eliminating ROS and reducing caspase-3 activity.