Cellular Responses to Acetamiprid in Human Granulosa Cells: Molecular Docking Insights Into PI3K/AKT Signaling Pathway

ABSTRACT

Acetamiprid (ACE) is a widely used neonicotinoid insecticide, and increasing evidence suggests potential adverse effects on reproductive‐related cellular functions; however, its impact on human ovarian granulosa cells is not yet fully clarified. In this study, human granulosa cells (HGrC1) were exposed to ACE, and its effects on cell viability, migration, apoptosis‐related markers, and stress‐associated molecular pathways were evaluated. Cell viability analysis demonstrated a concentration‐ and time‐dependent decrease following ACE exposure. In addition, wound healing assays indicated a significant reduction in cell migration capacity and Annexin V/PI staining analysis revealed enhanced apoptotic activity. Molecular analyses revealed elevated expression of Caspase‐3 (Cas‐3) and Bax at both mRNA and protein levels, suggesting activation of apoptosis‐related signaling. Furthermore, elevated ATM and iNOS expression levels indicated the involvement of DNA damage response– and oxidative stress‐associated cellular processes. TOS/TAS analyses demonstrated increased oxidative stress following ACE exposure. Complementary molecular docking analysis suggested potential interactions between ACE and PI3K/AKT signaling proteins. Immunohistochemical analysis of p110 α , the catalytic subunit of PI3K, revealed a significant decrease in immunoreactivity following ACE exposure. Overall, these findings indicate that ACE exposure is associated with altered cellular responses in human granulosa cells, affecting viability, migration, apoptosis, and oxidative stress‐related signaling pathways, while in silico results provide additional insight into possible involvement of PI3K/AKT interactions. However, further experimental studies are necessary to confirm these molecular interactions and to fully elucidate the underlying mechanisms.