A systems perspective on ribavirin-induced hemolytic anemia: From network toxicology to atomic-level simulations

The molecular mechanisms underlying ribavirin (RBV)-induced hemolytic anemia remain incompletely understood. Current research predominantly addresses individual targets or pathways, lacking a comprehensive integration of multi-target interaction networks. This study employs a combination of network toxicology (NT), multi-scale molecular dynamics (MD) simulation techniques, and in vitro experiments to conduct an in-depth analysis of the multi-target mechanisms responsible for RBV-induced hemolytic anemia. Methods: A multi-scale biological molecular network model was developed utilizing NT. Multi-scale molecular simulations were employed to integrate calculations across various temporal and spatial scales. This approach was complemented by red blood cell hemolysis assays, liquid chromatography-tandem mass spectrometry (LC-MS/MS) quantitative analysis, and circular dichroism (CD) spectroscopy experiments, thereby elucidating the hierarchical patterns of molecular behaviors that contribute to RBV-induced hemolytic anemia across multiple scales.Results: The findings demonstrated that RBV/triphenyl ribavirin (RTP) disrupts the function of the Band 3 protein, inhibits glucose-6-phosphate dehydrogenase (G6PD) activity, induces oxidative stress, and triggers apoptotic pathways, ultimately resulting in the rupture of red blood cells. The integration of multi-scale data corroborated the "drug accumulation-oxidative stress-cell death" three-stage damage model, offering a quantitative characterization framework from atomic to systemic levels for elucidating the toxicity mechanism of nucleoside analogues. This approach advances the application of analytical toxicology in the evaluation of drug safety.