The Effect of Danshen on Fibrosis and Iron Metabolism in a Rat Model of Hepatic Fibrosis

Background: Hepatic fibrosis (HF) is a major global health burden with limited effective therapies. Ferroptosis, an iron-dependent form of cell death, and inflammatory pathways are implicated in HF progression. Salvia miltiorrhiza (Danshen) has demonstrated antifibrotic effects, but its underlying mechanisms remain unclear. Objective: To elucidate the mechanisms of Danshen in treating HF through network pharmacology and experimental validation, focusing on the JAK/STAT pathway, iron metabolism, and ferroptosis. Methods: Using the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database, we identified 92 common targets among Danshen components, HF-related genes, and iron metabolism genes. A CCl4-induced HF rat model was established (n = 70) and treated with Danshen (3 or 6 g/kg/d) for 4 weeks. Hepatic fibrosis was assessed by Masson staining. Serum markers including Fe2+, transferrin (TRF), ferritin (SF), type III procollagen (PCIII), hydroxyproline (HyP), malondialdehyde (MDA), and glutathione (GSH) were measured. Western blotting evaluated hepatic expression of IL-6, P-JAK2, p-STAT3, and GPX4. Results: Network pharmacology revealed JAK2/STAT3 as top hub genes among 92 intersection targets. High-dose Danshen decreased IL-6, P-JAK2, and p-STAT3 expression by 45–60% (p < 0.05), increased serum Fe2+ and TRF by 1.8-fold, and reduced SF, HyP by >40% and PCIII by >15% (p < 0.05). Additionally, Danshen restored GPX4 protein expression and suppressed ferroptosis by decreasing MDA while increasing GSH (p < 0.01). Masson staining showed 55% reduction in collagen deposition with high-dose treatment. Conclusion: Danshen alleviates hepatic fibrosis by modulating the JAK/STAT-iron-ferroptosis axis, establishing a novel multi-target therapeutic strategy for HF.