Di(2‐Ethylhexyl) Phthalate Exposure and Risk of Diabetic Kidney Disease: Epidemiological Association and Mechanistic Insights

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

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.

Association between TBPH exposure, increased MMP9 expression, and atherosclerosis: evidence from integrated network analysis and in vivo experiments

Bis(2-ethylhexyl)-2,3,4,5-tetrabromophthalate (TBPH) is used as a nonreactive brominated flame retardant used in industrial products, particularly in flexible polyvinyl chloride and rigid and flexible polyurethane foams. Although TBPH exposure poses potential health risks, its association with atherosclerosis remains poorly understood. This study integrated network toxicology, machine learning, molecular docking, and molecular dynamics (MD) simulations to elucidate the influence of TBPH on high-fat diet–induced atherosclerosis. TBPH exhibited significant predicted toxicity across multiple organs. In total, 167 potential TBPH targets and 690 atherosclerosis-associated targets were identified, with 108 overlapping targets obtained through intersectional analysis. A PPI network was constructed based on these 108 overlapping genes, and the top 20 hub genes were identified. In a parallel independent analysis using a public transcriptomic dataset, 111 differentially expressed genes (DEGs) were identified by comparing atherosclerotic and normal tissues. Subsequent intersection analysis of the 108 overlapping targets and 111 DEGs identified candidate genes that were both TBPH-related and differentially expressed in atherosclerosis. Public transcriptomic data showed higher matrix metalloproteinase 9 (MMP9) level in atherosclerotic tissues. The corresponding receiver operating characteristic (ROC) curve yielded an AUC of 0.806 (95% confidence interval [CI]: 0.698–0.914). Molecular docking and MD simulations indicated favorable predicted binding modes and computational stability between TBPH and MMP9. In vivo experiments further demonstrated an association between TBPH exposure, increased MMP9 expression, and high–fat diet–induced atherosclerotic lesions. These findings suggest that TBPH may be associated with high-fat diet–induced atherosclerosis progression through the dysregulation of key mediators, providing insight into environmental pollutant–driven disease development.

Association between serum perfluoroalkyl substances and COPD risk: population evidence and in vitro validation.

Mixture exposure showed a significant positive association with chronic obstructive pulmonary disease risk, and these substances disrupt pulmonary homeostasis through concurrent molecular activation and lipid-metabolic disturbance, evidenced by triglyceride-glucose index mediation.

Topical exposure to di(2-ethylhexyl) phthalate aggravates atopic dermatitis: an integrated network toxicology, bioinformatics, and experiment analysis.

Di(2-ethylhexyl) phthalate (DEHP) is one of the most widely used plasticizers; however, whether cutaneous DEHP exposure affects atopic dermatitis (AD) remains unclear. We combined network toxicology, transcriptomic analysis, machine learning, in vivo and in vitro experimental validation, and molecular docking to investigate whether cutaneous DEHP exposure exacerbates AD. ADMETlab 3.0 profiling of DEHP and its major metabolites predicted skin sensitization as one of the higher-probability toxicity endpoints among the evaluated computational outputs. We subsequently intersected DEHP-related targets with AD-associated genes and identified 62 shared targets. The shared targets were enriched in biological processes related to T-cell proliferation, cell adhesion and leukocyte extravasation, as well as inflammatory pathways including JAK-STAT, MAPK and PI3K-Akt signaling. Using STRING and Cytoscape, we constructed a "DEHP-shared targets-pathways-AD" network and prioritized eight hub genes for further investigation. Bulk transcriptomic analysis, together with feature-prioritization results from five machine-learning models, supported the relevance of most candidate genes, six of which were significantly upregulated in AD lesions. Importantly, in vivo experiments demonstrated that topical DEHP exposure exacerbates MC903-induced AD-like dermatitis in BALB/c mice. This exacerbation was accompanied by increased expression of vascular and immune-related genes, including SELE, SELP, CXCR1, and ITGAL. Immunohistochemical staining further supported this vascular-immune activation pattern, revealing increased SELE and SELP expression in ear skin lesions in the MC903 + DEHP group, compared with MC903 + vehicle group. In vitro experiments also showed that DEHP stimulation upregulated SELE and SELP expression in endothelial cells. Collectively, these findings implicate endothelial adhesion, leukocyte migration, and immune-cell activation in DEHP-associated exacerbation of AD-like inflammation. Finally, molecular docking suggested potential binding modes between DEHP and these proteins, providing structural hypotheses for their possible involvement in this process. Together, our findings indicate that cutaneous DEHP exposure may exacerbate AD by promoting endothelial adhesion and immune-cell infiltration, thereby intensifying skin inflammation.

Organophosphate flame retardants and incident type 2 diabetes: Integrating population evidence, mechanistic insights, and effect modification by healthy lifestyle.

Mixed OPFRs exposure, dominated by TPHP, was associated with higher incident T2DM risk, partly attenuated by healthier lifestyle trajectories, supporting combined exposure-reduction and lifestyle-based prevention strategies.

Toxicological Effects of Propyl 4-Hydroxybenzoate on Gallstone Pathogenesis: An Integrated Mendelian Randomization, Network Toxicology, and Experimental Study.