器官移植2025，Vol.16Issue(5)：718-727,10.DOI:10.12464/j.issn.1674-7445.2025141
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
Silencing FABP4 alleviated ferroptosis of renal tubular epithelial cells induced by hypoxia/reoxygenation through regulating Nrf2/GPX4 axis
摘要
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.关键词
脂肪酸结合蛋白4/核因子E2相关因子2/谷胱甘肽过氧化物酶4轴/缺氧/复氧/肾小管上皮细胞/铁死亡/细胞增殖活性/活性氧簇/急性肾损伤Key words
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分类
医药卫生引用本文复制引用
白冰,谭州科,石彬,蒲涛,杨亦彬..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[J].器官移植,2025,16(5):718-727,10.基金项目
国家自然科学基金地区科学基金项目(82360154) （82360154）
贵州省科技计划项目(黔科合基础-ZK[2023]一般568) （黔科合基础-ZK[2023]一般568）
遵义市科技与大数据项目[遵市科合HZ字(2022)358号] （2022）
