Silencing FABP4 alleviated ferroptosis of renal tubular epithelial cells induced by hypoxia/reoxygenation through regulating Nrf2/GPX4 axis
10.12464/j.issn.1674-7445.2025141
- VernacularTitle:沉默FABP4通过调节Nrf2/GPX4轴减轻缺氧/复氧诱导的肾小管上皮细胞铁死亡
- Author:
Bing BAI
1
;
Zhouke TAN
;
Bin SHI
;
Tao PU
;
Yibin YANG
Author Information
1. 563000 贵州遵义,遵义医科大学附属医院肾内科
- Publication Type:Journal Article
- Keywords:
Fatty acid binding protein 4;
Nuclear factor E2-related factor 2/glutathione peroxidase 4 axis;
Hypoxia/reoxygenation;
Renal tubular epithelial cell;
Ferroptosis;
Cell proliferation activity;
Reactive oxygen species;
Acute kidney injury
- From:
Organ Transplantation
2025;16(5):718-727
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects and mechanisms of fatty acid binding protein 4(FABP4)on ferroptosis in human renal tubular epithelial cells(HK-2)treated with hypoxia/reoxygenation(H/R).Methods HK-2 cells were cultured in vitro and subjected to hypoxia for 24 hours followed by reoxygenation for different durations(1,3,6 h).The messenger RNA(mRNA)and protein levels of FABP4 in HK-2 cells were detected at each time point using real-time fluorescent quantitative polymerase chain reaction and Western blotting.Small interfering RNA(siRNA)technology was used to silence the expression of FABP4 gene in HK-2 cells,which were then treated with H/R(24 h of hypoxia and 6 h of reoxygenation)or treated with the Nrf2 inhibitor ML385.Cell proliferation activity was assessed using cell counting kit-8(CCK-8).Lactate dehydrogenase(LDH)levels were measured by enzyme-linked immune absorbent assay.Malondialdehyde(MDA),glutathione(GSH)and ferrous ion(Fe2+)levels were determined by biochemical technology.Reactive oxygen species(ROS)levels were detected using the 2',7'-dichlorodihydrofluorescein diacetate fluorescence probe.Protein expression levels of FABP4,nuclear factor E2-related factor 2(Nrf2),heme oxygenase-1(HO-1),glutathione peroxidase 4(GPX4)and solute carrier family 7 member 11(SLC7A11)were measured by Western blotting.Results The mRNA and protein levels of FABP4 in HK-2 cells increased with prolonged reoxygenation time(all P<0.05).H/R treatment reduced cell proliferation activity,increased LDH levels in the cell supernatant,and elevated MDA,Fe2+and ROS levels in HK-2 cells while decreasing GSH levels and the protein levels of Nrf2,HO-1,GPX4 and SLC7A11(all P<0.05).Silencing FABP4 enhanced the proliferation activity of H/R-treated HK-2 cells(P<0.05),reduced MDA,Fe2+and ROS levels,increased GSH levels,and elevated the protein levels of Nrf2,HO-1,GPX4 and SLC7A11(all P<0.05).However,these beneficial effects of FABP4 silencing on H/R-induced ferroptosis in HK-2 cells were reversed by co-treatment with ML385.Conclusions Silencing FABP4 alleviated H/R-induced ferroptosis in HK-2 cells,possibly by activating the Nrf2/GPX4 axis.
