Roumudan decoction attenuates hepatic fibrosis by suppressing the Warburg effect via inhibiting key glycolytic proteins.

[ETHNOPHARMACOLOGICAL RELEVANCE] Hepatic fibrosis serves as a critical pathological intermediate in the progression of chronic liver disease to liver cirrhosis and ultimately to hepatocellular carcinoma, posing a substantial threat to human health. Currently, clinically available therapies for hepatic fibrosis are hampered by limitations, including suboptimal efficacy and a high incidence of adverse effects. Roumudan (RMD), a classic formulation for liver disease management at the Affiliated Hospital of Shaanxi University of Chinese Medicine, has demonstrated therapeutic efficacy in hepatic fibrosis through rigorous long-term clinical validation, yet its precise mechanistic underpinnings remain elusive.

[AIM OF THE STUDY] This study seeks to systematically assess the therapeutic potential of RMD in hepatic fibrosis and unravel its underlying mode of action, with a particular focus on its regulatory impact on the Warburg effect (aerobic glycolysis).

[MATERIALS AND METHODS] A rat hepatic fibrosis model was induced through intraperitoneal CCL injection. RMD's therapeutic impact was evaluated using various parameters, such as body weight, liver function, inflammatory state, oxidative stress, HSC activation, and ECM dynamics. UPLC-HRMS was utilized to detect the absorbed components of RMD. Network pharmacology predicted potential targets, which were validated via transcriptome sequencing, RT-qPCR, immunofluorescence, and Western blotting.

[RESULTS] RMD dose-dependently attenuated fibrotic indices including liver injury, inflammation, oxidative stress, and collagen accumulation. Pharmacokinetic analysis identified 42 bioactive RMD constituents. Bioinformatics revealed 181 overlapping targets significantly enriched in glycolytic pathways (P < 0.05). RMD suppressed transcription and expression of core glycolytic regulators GLUT1, HK2, PKM2, and LDH-A, disrupting HSC metabolic reprogramming.

[CONCLUSION] RMD inhibits hepatic fibrogenesis through multi-targeted suppression of Warburg-associated glycolysis, establishing its mechanistic foundation and nominating glycolytic enzymes as therapeutic targets.