Triazole fungicides are highly effective pesticides widely used in plant protection, which has caused potential hazards to human health and ecological safety. To fully understand their potential hepatotoxicity, we first analyzed the transcriptome profiles in HepG2 cells treated with five triazole fungicides (hexiconazole (HEX), tebuconazole (TEB), propiconazole (PRO), cyproconazole (CYP), and difenoconazole (DIF)), and found that these pesticides remarkably affected estrogen signaling pathways, especially estrogen synthesis. Furthermore, we found that TEB, CYP, PRO and DIF had agonistic activity towards estrogen receptor alpha (ERα) and elucidated the binding mode of triazole ligands with ERα using the reporter gene assay and molecular docking. Four triazole fungicides regulated eight major genes involved in estrogen synthesis (StAR, CYP11A1, 3βHSD2, CYP17, CYP19, CYP3A4, CYP1A2 and SCP2) and stimulated the secretion of 17β-estradiol (E2). Finally, we assessed possible metabolic outcomes caused by abnormal estrogen synthesis, and found that triazole fungicides affected the metabolism of various macromolecules (such as lipid, amino acid, and carbohydrate) and signal transduction. These findings will provide new insights into endocrine-disrupting effects of triazole fungicides and highlight their potential ecological and health risks.