Exploring the Molecular Mechanisms by Which DEHP Promotes the Occurrence and Progression of Kidney Stones Based on Network Toxicology and Experimental Validation

Abstract

Background The global burden of kidney stones is increasing, while di(2-ethylhexyl) phthalate (DEHP), a widespread plasticizer and endocrine-disrupting pollutant, has been implicated as a potential environmental risk factor. This study investigated the mechanisms by which DEHP may aggravate calcium oxalate kidney stones through network toxicology, computational analyses, and experimental validation. Methods DEHP- and kidney stone–related targets were retrieved from six databases. Shared targets were analyzed using protein–protein interaction networks and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses. Molecular docking and 100-ns molecular dynamics simulations were performed to evaluate interactions between DEHP and hub proteins. A glyoxylic acid–induced mouse model and calcium oxalate monohydrate (COM)-challenged HK-2 cells were used to assess the effects of DEHP on crystal deposition, renal injury, apoptosis, and autophagy. Results A total of 457 shared targets were identified, with CTNNB1, SRC, HSP90AA1, EP300, EGFR, and TP53 emerging as hub genes. Computational analyses suggested stable interactions between DEHP and these proteins and highlighted cell fate–related pathways, including PI3K–Akt signaling, autophagy, and apoptosis. In vivo, DEHP increased blood urea nitrogen and serum creatinine levels, aggravated tubular injury, and enhanced calcium oxalate crystal deposition. Consistent effects were observed in COM-treated HK-2 cells, accompanied by dysregulation of apoptosis- and autophagy-related markers. Conclusion DEHP aggravates calcium oxalate crystal–induced renal injury and promotes kidney stones, potentially by disrupting apoptosis–autophagy homeostasis in renal tubular epithelial cells. These findings provide mechanistic evidence linking DEHP exposure to kidney stone pathogenesis.