Pien Tze Huang ameliorates alcohol-associated liver disease via suppressing oxidative stress and ferroptosis.

Abstract

BACKGROUND: Alcohol-associated liver disease (ALD) is a major public health concern due to its increasing prevalence and lack of effective therapies. Pien Tze Huang (PTH) is a traditional Chinese medicinal formula commonly used for viral hepatitis and liver fibrosis. However, whether it also has a robust therapeutic effect on ALD remains unclear. PURPOSE: This study aimed to verify the effect of PTH on ALD, identify the active components, and explore the underlying mechanisms. METHODS: HPLC-Q-TOF-MS and GC-MS were performed for chemical constitution analysis, quality control, and bioactive components identification. An ALD mouse model was established via gavage of 50% ethanol, while oxidative stress and ferroptosis in HepG2 cells were induced using ethanol or tert‑Butyl hydroperoxide. Biochemical indicators and pathological analysis were conducted to evaluate the effects. Transcriptome, polymerase chain reaction, western blotting, and siRNA/plasmids transfection were performed to explore and verify the mechanism. RESULTS: PTH significantly attenuated ALD in mice. It had marginal effects on alcohol metabolism but strongly suppressed oxidative stress and ferroptosis. Mechanistically, PTH restored Keap1-Nrf2 signaling and inhibited ferric iron import. Calculus Bovis (from Bos taurus domesticus) and Musk (Moschus, from Moschus berezovskii, M. sifanicus, or M. moschiferus) were bioactive ingredients. Bovinic acid and taurine (1.3 and 2.3 mg/g in PTH, respectively) were bioactive components. They directly bound SIRT1 at different amino acid residues to suppress oxidative stress and ferroptosis via SIRT1/Keap1/Nrf2 axis and SIRT1/transferrin receptor signal. CONCLUSIONS: Bovinic acid and taurine in PTH suppress SIRT1/Keap1/Nrf2 axis-mediated oxidative stress and SIRT1/ transferrin receptor-mediated ferroptosis to ameliorate ALD.