Ochratoxin A perturbs reproductive cell homeostasis involving PI3K-Akt-related stress signaling: Network toxicology, molecular simulation, single-cell and in vitro evidence.

Abstract

Ochratoxin A (OTA) is a persistent foodborne environmental contaminant with recognized systemic toxicity, but the molecular basis of its reproductive effects remains incompletely defined. We integrated network toxicology, molecular docking, molecular dynamics simulation and public single-cell RNA sequencing to generate candidate mechanisms, and then evaluated selected predictions using cell-viability and transcript-level assays. Intersections between OTA-related and reproductive-injury targets yielded 318 testicular and 426 ovarian candidate targets. AKT1, MAPK1, PIK3CA, SRC and TP53 were shared network hubs, while enrichment analysis prioritized PI3K-Akt signaling, apoptosis, p53, FoxO and MAPK pathways. Docking predicted favorable OTA poses for the five hub proteins, and 100-ns simulations of the selected AKT1-OTA and PIK3CA-OTA complexes showed persistence of the predicted poses within the modeled systems; these computational findings do not establish biological binding or direct target engagement. Public single-cell data localized the candidate genes to germ, somatic and steroidogenic cell populations. In vitro, OTA reduced 24-h viability with IC50 values of 9.25 micromolar in GC-1 cells and 27.94 micromolar in KGN cells and altered hub-gene transcripts. Taken together, the results suggest that OTA-associated reproductive cell stress may involve PI3K-Akt-related survival signaling and apoptosis-linked responses, but protein-level and functional validation is required.