Screening Hub Genes of Ferroptosis in Pb-Induced Acute Kidney Injury Based on GEO Database Bioinformatics and Experimental Validation

J Appl Toxicol. 2025 Dec;45(12):2688-2699. doi: 10.1002/jat.4866. Epub 2025 Jul 25.

Abstract

This study aimed to explore the role of ferroptosis-related genes in Pb-induced acute kidney injury (AKI) using bioinformatics and animal experiments, potentially providing a new direction for future research. Differentially expressed genes related to Pb-induced AKI mediated by ferroptosis were identified using databases. Subsequently, hub genes were identified through enrichment and protein-protein interaction analyses. Western blot and immunohistochemistry were used to verify the expression of hub genes and ferroptosis marker protein GPX4, and the levels of Fe2+, GSH, and MDA in tissues were determined. Drug prediction was performed using the DSigDB database and molecular docking. The hub genes were screened: heme oxygenase 1 (HMOX1), CD44 molecule (CD44), arachidonate 15-lipoxygenase (ALOX15), and NADPH quinone dehydrogenase 1 (NQO1). In 600 mg/kg lead acetate-induced AKI rat models, kidney tissues exhibited significantly lower levels of HMOX1, CD44, NQO1, and glutathione peroxidase 4 (GPX4) proteins, while showing higher levels of ALOX15 protein. Furthermore, significantly elevated levels of Fe2+ and malondialdehyde were observed, accompanied by decreased levels of reduced glutathione; potential therapeutic drugs may include resveratrol, retinoic acid, valproic acid, and tert-butylhydroquinone. Our study reveals an association between Pb exposure and the dysregulation of ferroptosis-related genes (HMOX1, CD44, ALOX15, and NQO1). We hypothesize that these genetic alterations may contribute to ferroptosis and subsequent AKI. Further research with larger sample sizes and additional experimental validation is needed.

Keywords: Pb; acute kidney injury; bioinformatics; drug prediction; ferroptosis.

MeSH terms

  • Acute Kidney Injury* / chemically induced
  • Acute Kidney Injury* / genetics
  • Acute Kidney Injury* / metabolism
  • Animals
  • Arachidonate 15-Lipoxygenase / genetics
  • Arachidonate 15-Lipoxygenase / metabolism
  • Computational Biology
  • Ferroptosis* / drug effects
  • Ferroptosis* / genetics
  • Heme Oxygenase-1 / genetics
  • Heme Oxygenase-1 / metabolism
  • Hyaluronan Receptors / genetics
  • Hyaluronan Receptors / metabolism
  • Kidney / drug effects
  • Kidney / metabolism
  • Kidney / pathology
  • Lead* / toxicity
  • Male
  • Molecular Docking Simulation
  • NAD(P)H Dehydrogenase (Quinone) / genetics
  • Phospholipid Hydroperoxide Glutathione Peroxidase / genetics
  • Phospholipid Hydroperoxide Glutathione Peroxidase / metabolism
  • Protein Interaction Maps
  • Rats
  • Rats, Sprague-Dawley

Substances

  • Lead
  • Arachidonate 15-Lipoxygenase
  • Phospholipid Hydroperoxide Glutathione Peroxidase
  • NAD(P)H Dehydrogenase (Quinone)
  • Heme Oxygenase-1
  • Hyaluronan Receptors