Background: alpha-Amanitin, the primary lethal toxin of Amanita phalloides, induces irreversible hepatotoxicity by selectively inhibiting RNA polymerase II, leading to transcriptional arrest. Despite advancements in managing mushroom poisoning, a targeted antidote remains unavailable. The sodium taurocholate co-transporting polypeptide (NTCP), a hepatic bile acid transporter, facilitates alpha-amanitin entry into hepatocytes. Pharmacological blockade of NTCP represents a promising therapeutic strategy. Objective: To evaluate ezetimibe, an NTCP inhibitor, as a protective agent against alpha-amanitin-induced hepatotoxicity. Methods: Transcriptomic profiling of alpha-amanitin-exposed mouse liver tissues (NCBI accession: PRJNA809431) was conducted using DESeq2. Molecular docking simulations assessed interactions between NTCP, alpha-amanitin, and ezetimibe. Therapeutic efficacy was evaluated in vivo (mouse models) and in vitro (cultured hepatocytes). Key outcomes included survival rates, liver injury markers (ALT, AST), apoptosis (Bax/Bcl-2 ratio), and oxidative stress parameters. Results: NTCP expression was upregulated in alpha-amanitin-exposed livers. Molecular docking revealed alpha-amanitin binding at NTCP residue VAL-160, whereas ezetimibe interacted with LEU-14 and ASN-17. Ezetimibe (50 mg/kg) improved survival rates from 25 % to 80 % in alpha-amanitin-exposed mouse models (p < 0.01), reduced serum ALT (68 f 5 U/L vs. 165 f 12 U/L; p < 0.05) and AST (72 f 6 U/L vs. 158 f 10 U/L; p < 0.05), and attenuated apoptosis (60 % decrease in Bax/Bcl-2; p < 0.05). In vitro, ezetimibe restored hepatocyte viability 2.1-fold (p < 0.05) and reduced oxidative stress (40 % decrease in malondialdehyde; p < 0.05). Transcriptomic analysis linked alpha-amanitin toxicity to p53-mediated apoptosis. Conclusion: Ezetimibe protects against alpha-amanitin hepatotoxicity by blocking NTCP-mediated uptake, supporting its potential clinical repurposing as a targeted antidote.