Multi-level bioinformatics combined with in vitro and in vivo experiments reveal the nephrotoxicity risk of isotoosendanin

Fructus Meliae Toosendan (FMT) has been extensively employed in the field of biopesticides due to its significant insecticidal properties and has consequently emerged as an innovative biopesticide. Isotoosendanin (ITSN), a limonoid from FMT. Although ITSN is regarded as the principal toxic constituent within FMT, the underlying toxicological mechanisms of ITSN have yet to be fully elucidated. This study assesses the nephrotoxic effects of ITSN through a multi-level bioinformatics analysis, integrated with both in vitro and in vivo experimental approaches. Using network toxicology, 116 nephrotoxicity-related targets were predicted, including 10 core targets such as CASP3, EGFR, and MTOR. Molecular docking and dynamics simulations showed strong affinity and stability between ITSN and these targets. In vitro experiments on HK-2 renal epithelial cells demonstrated that ITSN inhibits proliferation and migration, induces apoptosis, and increases ROS and inflammatory cytokines (TNF-α, IL-1β) in a dose-dependent manner, suggesting oxidative stress and inflammation as key toxic effects. In vivo studies conducted on murine models demonstrated that the group receiving a dosage of 30 mg/kg exhibited increased serum concentrations of creatinine and urea. Additionally, this group showed structural damage to the glomeruli and tubules, accompanied by interstitial inflammatory infiltration. Mechanistically, ITSN-induced nephrotoxicity is intricately linked to the suppression of the PI3K/AKT signaling pathway and the subsequent induction of apoptosis. The administration of the PI3K agonist 740Y-P markedly enhances cell viability. These findings enhance our understanding of ITSN's toxic potential, supporting its safe use in medical and agricultural applications and contributing to toxicity assessment strategies for natural compounds.