Glyphosate promotes calcium oxalate crystal-induced renal injury by modulating the PI3K/Akt-mediated mechanism

The increasing global prevalence of nephrolithiasis represents a significant public health concern. However, the role of environmental contaminants such as glyphosate remains unclear. Herein, epidemiological analysis, network toxicology, and experimental validation were integrated to investigate this association. Cross-sectional analysis of NHANES data demonstrated a significant dose-dependent relationship between urinary glyphosate levels and nephrolithiasis prevalence. Each unit increase in log-transformed glyphosate concentration was associated with a 25% increase in nephrolithiasis risk (OR = 1.25, 95% CI: 1.06–1.47). Network toxicology combined with molecular docking and dynamics simulations identified the PI3K/AKT signaling pathway as a potential target, showing stable binding of glyphosate to core proteins. This mechanism was functionally validated in vivo and in vitro, indicating that glyphosate co-exposure exacerbated calcium oxalate-induced renal tubular injury, oxidative stress, and apoptosis through suppression of the PI3K/AKT pathway. This effect was significantly reversed by pathway agonism. Collectively, this study provides the first integrative evidence identifying environmental glyphosate exposure as a novel risk factor for nephrolithiasis. Furthermore, inhibition of the PI3K/AKT pathway was delineated as the primary underlying mechanism, offering a conceptual framework for future preventive and therapeutic strategies. • Urinary glyphosate levels are dose-dependently associated with nephrolithiasis risk. • Glyphosate promotes CaOx crystal-induced renal injury by inhibiting PI3K/AKT (confirmed in vivo, in vitro & network toxicology), and pathway agonism reverses this damage. • First evidence links glyphosate to nephrolithiasis via PI3K/AKT suppression, offering novel therapeutic targets.