Background: Hydroxysafflor yellow A (HSYA), isolated from the dried flower of Carthamus tinctorius L., has gained extensive attention due to its broad and effective pharmacological activities. However, its antifibrotic effects on liver fibrosis, with mainly focusing on hepatic stellate cells (HSCs) as the key cells in liver fibrogenesis, are still completely unverified. Objective: this study provides insights on the exact mechanism of actions of HSYA on HSCs both in vitro and in thioacetamide (TAA)-induced liver fibrosis. Materials and Methods: The in vitro study was performed on stellate cell line (HSC-T6) whereas liver fibrosis was established in rats via chronic TAA intoxication. TAA-intoxicated rats were treated with silymarin (50 mg/kg) or HSYA (5 mg/kg) daily for 8 weeks via oral gavage. Serum liver enzymes, proinflammatory cytokines, fibrosis markers, expression of proliferating cellular nuclear antigen (PCNA) together with histopathological changes and immunohistochemical staining were examined. Results: HSYA showed only a minor suppressor effect on HSCs proliferation, which did not exceed 9.5% reduction, in a concentration of 500 µg/mL at 48 h. However, it significantly reduced TGF-β1, inhibited α-SMA expression (88.73%), induced caspase-3 expression (5.3 fold vs untreated cells) and showed flattened cell morphology (senescence). HSYA also ameliorated liver functions, suppressed IL-6 and TNF-α, mitigated hepatic levels of PDGF-BB, TGF-β1, TIMP-1 and hydroxyproline with attenuation of fibrosis score (S2 vs S4). Additionally, HSYA down-regulated α‑SMA, restored PCNA and increased caspase-3 expression. Conclusion: Our findings provide mechanistic insights into the possible applicability of HSYA as a new therapeutic approach for slowing down or reversing the progression of hepatic fibrosis, given that no effective anti-fibrotic therapies are currently available.