Protection and Mechanism of Daucosterol Against Oxidative Stress Induced by Hydrogen Peroxide in Human Renal Tubular Epithelial Cells
10.11969/j.issn.1673-548X.2024.10.026
- VernacularTitle:胡萝卜苷对H2O2诱导的HK-2细胞氧化应激的保护作用研究
- Author:
Yingying JIANG
1
;
Liangen LIN
;
Wang LV
Author Information
1. 325000 温州市人民医院急诊科
- Keywords:
Daucosterol;
Oxidative stress;
Renal tubular epithelial cells;
Hydrogen peroxide;
Diabetic Nephropathy
- From:
Journal of Medical Research
2024;53(10):144-148
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the protective effect and mechanism of Daucosterol on H2O2-induced oxidative stress in hu-man renal duct epithelial cells(HK-2 cells).Methods HK-2 cells were cultured in vitro,and treated with different concentrations of H2O2(1 μmol/L,5μmol/L,10μmol/L,50μmol/L and 100pmol/L)for 24h in H2O2 treatment group;In Daucosterol intervention group,HK-2 cells were pretreated with 50μmol/L Daucosterol for 2h and then treated with different concentrations of H2O2.CCK-8 assay was used to detect cell viability in each group.Hochest33342/PI fluorescence staining and caspase-3 immunofluorescence assay were used to analyze cell apoptosis.The levels of reactive oxygen species(ROS)were analyzed using DCFH-DA fluorescent probe stai-ning.Western blot assay was used to analyze the expression levels of Bax,Bcl-2 and NOX4.Results The inhibition rates in 50μmol/L and 100μmol/L H2O2 treatment groups were 20.74％±3.32％and 53.90％±2.37％;the inhibition rates in Daucosterol intervention group were 10.29％±1.69％(t=4.859,P=0.008)and 37.38％±3.95％(t=6.212,P=0.003),respectively,and the differ-ences were statistically significant.The results of Hochest33342/PI fluorescence staining showed significantly more Hochest33342 fluores-cence signal aggregation and PI fluorescence positive cells in H2O2 treatment group than in Daucosterol intervention group.Immunofluores-cence assay showed that the expression level of caspase-3 in H2O2 treatment group was significantly higher than that in Daucosterol inter-vention group.ROS level in Daucosterol intervention group was significantly attenuated compared with H2O2 treatment group.The results of Western blot showed that the expression level of NOX4 protein in H2O2 treatment group was higher than that in the control group,but significantly decreased in Daucosterol intervention group;The expression level of Bax protein also decreased in Daucosterol intervention group,compared with H2O2 treatment group.Conclusion Daucosterol intervention can alleviate H2O2-induced oxidative stress in HK-2 cells and reduce apoptosis to exert a protective effect.
