What question did this study set out to answer?

This investigation aims to clarify the molecular mechanisms by which DEHP exposure leads to cardiac fibrosis.

March 2, 2026Open Access

DEHP exposure and cardiac fibrosis risk: An integrated network toxicology and molecular experimental investigation

Key Points

This investigation aims to clarify the molecular mechanisms by which DEHP exposure leads to cardiac fibrosis.
Utilized network toxicology to identify DEHP targets from databases.
Conducted molecular-dynamics simulations to study DEHP interactions with core targets.
Performed in vitro experiments on cardiomyocytes and cardiac fibroblasts to assess cellular responses to DEHP exposure.
Analyzed protein-protein interaction networks to reveal core regulatory hubs.
Identified a 12-hub protein interaction network involving SRC, STAT3, and EGFR.
Confirmed that DEHP exposure decreases cardiomyocyte viability and induces apoptosis.
Shown that DEHP promotes cardiac fibroblast differentiation into myofibroblasts, activating SRC and STAT3 phosphorylation.

Abstract

The environmental endocrine-disrupting chemical di-(2-ethylhexyl) phthalate (DEHP) has been epidemiologically linked to cardiac fibrosis, yet the underlying molecular mechanism remains poorly defined. To dissect the mechanism, we employed an integrative strategy combining network toxicology, molecular-dynamics (MD) simulations and in vitro experimental validation. Putative DEHP targets (n = 6439) were mined from the Comparative Toxicogenomics Database (CTD) and SwissTargetPrediction, while cardiac fibrosis–related genes (n = 2288) were curated from GeneCards and OMIM. Overlap analysis identified 1054 shared candidates. These were imported into STRING to construct a high-confidence protein–protein interaction (PPI) network, which was subsequently analyzed in Cytoscape, revealing a 12-hub core enriched in SRC, STAT3, EGFR and other key regulators. Functional enrichment underscored a prominent footprint in inflammatory responses and in the PI3K–Akt and MAPK signaling cascades. Molecular-docking analyses revealed high binding affinities between DEHP and the identified core targets, while MD simulations verified the structural stability of DEHP complexes with SRC, STAT3 and EGFR. In vitro, cardiomyocyte viability declined progressively with increasing DEHP concentrations. Exposure to 50 μg/mL DEHP elevated the BAX/BCL2 ratio, promoted cytochrome c release, and raised the number of TUNEL-positive cells, confirming that DEHP induces cardiomyocyte apoptosis. In cardiac fibroblasts, DEHP up-regulated transcription of Tgf-β1, Col1a1, and Acta2, driving their differentiation toward a myofibroblast phenotype. Western blot analyses revealed that DEHP activated SRC and STAT3 phosphorylation; pharmacologic inhibition of SRC or STAT3 phosphorylation attenuated DEHP-induced phenotypic conversion of cardiac fibroblasts. Collectively, DEHP exacerbates cardiac fibrosis by triggering cardiomyocyte apoptosis and by activating the SRC–STAT3 axis to promote the conversion of cardiac fibroblasts into myofibroblasts, providing clear evidence of DEHP-elicited cardiotoxicity and a theoretical basis for targeted intervention. • 1.Network toxicology analysis linked DEHP exposure to cardiac fibrosis and identified 12 core targets. • 2.Molecular docking and molecular dynamics confirmed that DEHP can stably bind to SRC, STAT3, and EGFR. • 3.DEHP can promote cardiomyocyte apoptosis and impair cell viability. • 4.DEHP can promote the transformation of cardiac fibroblasts into myofibroblasts by activating SRC and STAT3 phosphorylation.

DEHP exposure and cardiac fibrosis risk: An integrated network toxicology and molecular experimental investigation

Key Points

Abstract

Cite This Study