Research Paper Volume 16, Issue 7 pp 5987—6007

Identification and validation of the biomarkers related to ferroptosis in calcium oxalate nephrolithiasis

Chao Hou1, *, , Bing Zhong1, *, , Shuo Gu1, *, , Yunyan Wang1, , Lu Ji1, ,

  • 1 Department of Urology, The Affiliated Huai'an First People’s Hospital of Nanjing Medical University, Huai’an 223300, Jiangsu, China
* Equal contribution

Received: October 6, 2023       Accepted: February 20, 2024       Published: March 25, 2024
How to Cite

Copyright: © 2024 Hou et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Ferroptosis is a specific type of programmed cell death characterized by iron-dependent lipid peroxidation. Understanding the involvement of ferroptosis in calcium oxalate (CaOx) stone formation may reveal potential targets for this condition. The publicly available dataset GSE73680 was used to identify 61 differentially expressed ferroptosis-related genes (DEFERGs) between normal kidney tissues and Randall's plaques (RPs) from patients with nephrolithiasis through employing weighted gene co-expression network analysis (WGCNA). The findings were validated through in vitro and in vivo experiments using CaOx nephrolithiasis rat models induced by 1% ethylene glycol administration and HK-2 cell models treated with 1 mM oxalate. Through WGCNA and the machine learning algorithm, we identified LAMP2 and MDM4 as the hub DEFERGs. Subsequently, nephrolithiasis samples were classified into cluster 1 and cluster 2 based on the expression of the hub DEFERGs. Validation experiments demonstrated decreased expression of LAMP2 and MDM4 in CaOx nephrolithiasis animal models and cells. Treatment with ferrostatin-1 (Fer-1), a ferroptosis inhibitor, partially reversed oxidative stress and lipid peroxidation in CaOx nephrolithiasis models. Moreover, Fer-1 also reversed the expression changes of LAMP2 and MDM4 in CaOx nephrolithiasis models. Our findings suggest that ferroptosis may be involved in the formation of CaOx kidney stones through the regulation of LAMP2 and MDM4.


AUC: area under the ROC curve; BUN: blood urea nitrogen; CaOx: calcium oxalate; CDF: cumulative distribution function; DEFERGs: differentially expressed ferroptosis-related genes; DEGs: differentially expressed genes; EG: ethylene glycol; Fer-1: ferrostatin-1; GO: Gene Ontology; GSEA: gene set enrichment analysis; GSVA: gene set variation analysis; HE: hematoxylin and eosin; IHC: immunohistochemistry; KEGG: Kyoto Encyclopedia of Genes and Genomes; LASSO: least absolute shrinkage and selection operator; LPO: lipid peroxidation; PCA: principal component analysis; PPI: protein-protein interaction; ROC: receiver operating characteristic; ROS: reactive oxygen species; RPs: Randall’s plaques; TUNEL: terminal deoxynucleotidyl transferase mediated uridine triphosphate nick end labeling; UA: uric acid; VK: von Kossa staining; WGCNA: weighted gene co-expression network analysis.