ORIGINAL ABSTRACT

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.