Ou Zejin, Li Ying, Chen Shi, Wang Ziyi, He Meiyi, Chen Zhicheng, Tang Shihao, Meng Xiaojing, Wang Zhi
Key Laboratory of Occupational Environment and Health, Guangzhou Twelfth People's Hospital, Guangzhou 510620, China.
Institute of Occupational and Environmental Health, Guangzhou Medical University, Guangzhou 510620, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2025 Aug 20;45(8):1743-1750. doi: 10.12122/j.issn.1673-4254.2025.08.18.
To investigate the role of ferroptosis in diquat-induced acute kidney injury (AKI) and its molecular mechanisms.
Transgenic zebrafish models with Tg (Eco.Tshb:EGFP) labeling of the renal tubules and Tg (lyz:dsRed2) labeling of the neutrophils were both divided into control group, gentamicin (positive control) group, diquat poisoning group, ferroptosis inhibitor group. The indicators of kidney injury, inflammatory response, and ferroptosis were examined in the zebrafish, and the changes in expressions of voltage-dependent anion-selective channel protein 1 (VDAC1) and mitochondrial ferritin (FTMT) were detected using Western blotting.
AKI induced by diquat exhibited a significant dose-effect relationship, and the severity of injury was proportional to the exposure concentration. Diquat also caused marked oxidative stress and inflammatory responses in the zebrafish models. Rhodamine metabolism assay and HE staining revealed significantly declined glomerular filtration function of the zebrafish as diquat exposure concentration increased. Immunofluorescence staining highlighted significant changes in the expressions of ferroptosis markers GPX4 and FTH1 in zebrafish renal tissues following diquat exposure. In diquat-exposed zebrafish, treatment with ferrostatin-1, a ferroptosis inhibitor, obviously upregulated GPX4 and downregulated FTH1 expressions and improved the metabolic rate of glucan labeled with rhodamine B. Diquat exposure significantly upregulated the expression of VDAC1 and FTMT in zebrafish, and the application of ferrostatin-1 and VBIT-12 (a VDAC1 inhibitor) both caused pronounced downregulation of FTMT expression.
Ferroptosis is a critical mechanism underlying diquat-induced AKI, in which VDAC1 and FTMT play important regulatory roles, suggesting their potential as therapeutic target for AKI caused by diquat.
探讨铁死亡在百草枯诱导的急性肾损伤（AKI）中的作用及其分子机制。
将肾小管标记为Tg（Eco.Tshb:EGFP）、中性粒细胞标记为Tg（lyz:dsRed2）的转基因斑马鱼模型分为对照组、庆大霉素（阳性对照）组、百草枯中毒组、铁死亡抑制剂组。检测斑马鱼的肾损伤、炎症反应和铁死亡指标，采用蛋白质免疫印迹法检测电压依赖性阴离子选择性通道蛋白1（VDAC1）和线粒体铁蛋白（FTMT）表达的变化。
百草枯诱导的急性肾损伤呈现显著的剂量-效应关系，损伤严重程度与暴露浓度成正比。百草枯还在斑马鱼模型中引起明显的氧化应激和炎症反应。罗丹明代谢试验和苏木精-伊红染色显示，随着百草枯暴露浓度增加，斑马鱼的肾小球滤过功能显著下降。免疫荧光染色突出显示，百草枯暴露后斑马鱼肾组织中铁死亡标志物GPX4和FTH1的表达发生显著变化。在百草枯暴露的斑马鱼中，用铁死亡抑制剂铁抑素-1治疗可明显上调GPX4表达、下调FTH1表达，并提高罗丹明B标记葡聚糖的代谢率。百草枯暴露显著上调斑马鱼中VDAC1和FTMT的表达，应用铁抑素-1和VBIT-12（一种VDAC1抑制剂）均导致FTMT表达明显下调。
铁死亡是百草枯诱导急性肾损伤的关键机制，其中VDAC1和FTMT发挥重要调节作用，提示它们作为百草枯所致急性肾损伤治疗靶点的潜力。
