Study on role of glutathione peroxidase 4-dependent ferroptosis in diclofenac-induced injury of human kidney tubular epithelial cells
10.3760/cma.j.cn114015-20240802-00679
- VernacularTitle:谷胱甘肽过氧化物酶4依赖的铁死亡在双氯芬酸所致人肾小管上皮细胞损伤中的作用研究
- Author:
Shuifang CHEN
1
;
Hui CHEN
;
Xuemei CHEN
;
Qianwen ZHENG
;
Dong ZHENG
Author Information
1. 厦门市中医院药学部，厦门市中药制剂研发与转化重点实验室，厦门 361009
- Publication Type:Journal Article
- Keywords:
Diclofenac;
Acute kidney injury;
Ferroptosis;
Glutathione peroxidase 4;
Ferrostatin-1;
Human kidney tubular epithelial cells;
Molecular mechanisms of pharma
- From:
Adverse Drug Reactions Journal
2025;27(5):260-267
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the role of glutathione peroxidase 4 (GPX4)-dependent ferroptosis in diclofenac-induced kidney injury.Methods:Human kidney tubular epithelial cells (HK-2 cells) were cultured and then divided into 3 groups: control group, diclofenac group, and iron death inhibitor ferrostatin-1 (Fer-1) group. The same amount of 1% Fer-1 (final concentration 10 μmol/L) and phosphate buffered saline was respectively added to cells in the Fer-1 group and the other 2 groups. After 48 hours of culture, diclofenac 200 μmol/L was added to cells in the diclofenac group and the Fer-1 group. The cell viability of each group was detected by cell counting kit-8 (CCK-8). The cell cycle, apoptosis and intracellular reactive oxygen species (ROS) levels were detected by flow cytometry. The levels of intracellular iron ion, lactate dehydrogenase (LDH), malondialdehyde (MDA) and GPX4 were detected by enzyme-linked immunosorbent assay. The expression level of GPX4 was detected by Western blotting method.Results:Compared with the control group, the cell viability and G1 phase cell percentage of the diclofenac group were significantly lower, and compared with the diclofenac group, those were significantly higher (all P<0.05). The apoptosis rate of diclofenac group was significantly higher than that of the control group ( P<0.05), but there was no significant difference in apoptosis rate between Fer-1 group and diclofenac group ( P>0.05). Compared with the control group, the intracellular ROS, iron content, LDH, and MDA levels were significantly higherin the diclofenac group, while the expression level of GPX4 was lower (all P<0.05). However, the ROS, iron content, LDH, and MDA levels in the Fer-1 group were lower than those in the diclofenac group, while GPX4 expression was higher than that in the diclofenac group (all P<0.05). Conclusion:Diclofenac can induce ferroptosis in HK-2 cells and inhibiting the ferroptosis can alleviate cell injury, suggesting that GPX4-dependent ferroptosis may be involved in kidney injury induced by diclofenac.
