Improvement effect of chrysophanol on hydrogen peroxide-induced apoptosis of EA.hy926 cells and its mechanism
10.13481/j.1671-587X.20240604
- VernacularTitle:大黄酚对H2O2诱导EA.hy926细胞凋亡的改善作用及其机制
- Author:
Siqi LI
1
,
2
;
Guangdao CHEN
;
Qiyi ZENG
Author Information
1. 暨南大学粤港澳中枢神经再生研究院神经生物学系,广东 广州 510632
2. 南方医科大学珠江医院儿科,广东 广州 510282
- Keywords:
Chrysophanol;
Bronchopulmonary dysplasia;
Apoptosis-inducing factor;
Nuclear translocation;
Apoptosis;
Oxidative stress
- From:
Journal of Jilin University(Medicine Edition)
2024;50(6):1512-1518
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To discuss the effect of chrysophanol on hydrogen peroxide(H2O2)-induced oxidative damage of the EA.hy926 cells,and to clarify its therapeutic role in bronchopulmonary dysplasia(BPD)and related mechanism.Methods:The EA.hy926 cells were induced with 25,50,100,200,400,800,and 1 600 μmol·L-1 H2O2,and 8,16,32,64,128,and 256 μmol·L-1 chrysophanol.CCK-8 method was used to detect the viabilities of the EA.hy926 cells treated with different concentrations of H2O2 and chrysophanol.The cells were divided into control group,model group(200 μmol·L-1 H2O2),low dose of chrysophanol group(8 μmol·L-1 chrysophanol and 200 μmol·L-1 H2O2),and high dose of emodin group(256 μmol·L-1 chrysophanol and 200 μmol·L-1 H2O2).Western blotting method was used to detect the expression levels of apoptosis-inducing factor(AIF)protein in the cytoplasm and nucleus in various groups;immunofluorescence staining was used to detect the AIF nuclear translocation in the cells in various groups;kits were used to detect the activities of superoxide dismutase(SOD)and the levels of malondialdehyde(MDA),cysteinyl aspartate specific proteinase(Caspase)-8,and Caspase-9 in the cells in various groups.Results:Under different concentrations of H2O2,the viabilities of EA.hy926 cells showed an inverted S-shaped curve,with good cell viability,and the half-maximal inhibitory concentration(IC50)was 261.52 μmol·L-1.The cell model was induced by 200 μmol·L-1 H2O2 for 24 h.As the increaseing of concentration of chrysophanol,there was no significant change of the viability in the EA.hy926 cells(P>0.05),and interventions were performed using 8 and 256 μmol·L-1 chrysophanol.The Western blotting results showed that compared with control group,the expression level of AIF protein in the nucleus in model group was significantly increased(P<0.05),and the expression level of AIF protein in the cytoplasm was significantly decreased(P<0.05).Compared with model group,the expression levels of AIF protein in the nucleus in both low and high doses of chrysophanol groups were significantly decreased(P<0.05),and the expression level of AIF protein in the cytoplasm was significantly increased(P<0.05).The immunofluorescence staining results showed that AIF was less localized in the nucleus in the cells in control group.Compared with control group,the positive value of AIF nuclear translocation in model group was significantly increased(P<0.05);compared with model group,the positive values of AIF nuclear translocation in both low and high doses of chrysophanol groups were significantly decreased(P<0.05).Compared with control group,the activity of SOD in the cells in model group was significantly decreased(P<0.05),and the level of MDA was significantly increased(P<0.01).Compared with model group,the activities of SOD in the cells in low and high doses of chrysophanol groups were significantly increased(P<0.05),and the level of MDA was significantly decreased(P<0.05 or P<0.01).There were no significant differences in the levels of Caspase-8 and Caspase-9 in the cells among various groups(P>0.05).Conclusion:Chrysophanol improves the H2O2-induced apoptosis of the EA.hy926 cells by inhibiting the oxidative stress and AIF nuclear translocation,which may be beneficial for the treatment of BPD.
