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Qiweizhigan granule ameliorates metabolic dysfunction-associated steatohepatitis by modulating the galectin 3/tumor necrosis factor receptor-associated factor 6-mediated ferroptosis
Yu-Nuo Yang, Yun-Hong Sun, Ming-Zhe Zhu, Yi-Rong Wang, Meng Li, Kai Wang, Jing Ma, Li Zhang, Dan Hu, Wen-Jun Zhou, Guang Ji, Yan-Qi Dang
Yu-Nuo Yang, Yun-Hong Sun, Ming-Zhe Zhu, Yi-Rong Wang, Meng Li, Li Zhang, Wen-Jun Zhou, Guang Ji, Yan-Qi Dang, Institute of Digestive Diseases, China-Canada Center of Research for Digestive Diseases, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
Yun-Hong Sun, Ming-Zhe Zhu, School of Public Health, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
Kai Wang, Experiment Center for Science and Technology, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
Jing Ma, Dan Hu, Seventh People’s Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai 200137, China
Li Zhang, Wen-Jun Zhou, Guang Ji, Yan-Qi Dang, State Key Laboratory of Integration and Innovation of Classic Formula and Modern Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
Co-first authors: Yu-Nuo Yang and Yun-Hong Sun.
Co-corresponding authors: Guang Ji and Yan-Qi Dang.
Author contributions: Yang YN and Sun YH wrote the initial manuscript as co-first authors; Yang YN, Sun YH, Zhu MZ and Wang YR performed the research; Li M, Wang K and Ma J reviewed the manuscript; Zhang L, Hu D, Zhou WJ, Ji G, and Dang YQ designed the research study; Ji G and Dang YQ have played important roles in the manuscript preparation as co-corresponding authors; all of the authors read and approved the final version of the manuscript to be published.
Supported by National Natural Science Foundation of China, No. 82530124; Digestive Diseases Committee of the Chinese Association of Traditional Chinese Medicine-The Youth Empowerment Program, No. 202557-006; and State Key Laboratory of Integration and Innovation of Classic Formula and Modern Chinese Medicine, No. LSLSKL20240127.
Institutional animal care and use committee statement: The Experimental Animal Welfare and Ethics Committee of Shanghai University of Traditional Chinese Medicine granted approval for this study (No. PZSHTCM210625020).
Conflict-of-interest statement: All authors declare no conflict of interest in publishing the manuscript.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Data sharing statement: All datasets generated for this study are included in the article.
Corresponding author: Yan-Qi Dang, PhD, Institute of Digestive Diseases, China-Canada Center of Research for Digestive Diseases, Longhua Hospital, Shanghai University of Tra
ditional Chinese Medicine, Fenglin Road Subdistrict, Shanghai 200032, China.
yq_dang@shutcm.edu.cn
Received: November 14, 2025
Revised: January 6, 2026
Accepted: February 13, 2026
Published online: May 7, 2026
Processing time: 164 Days and 2 Hours
BACKGROUND
Qiweizhigan granule (QWZG) is employed in clinical settings for the treatment of metabolic dysfunction-associated steatohepatitis (MASH). However, the precise biological mechanisms underlying its therapeutic effects are not yet fully elucidated.
AIM
To assess the efficacy and the mechanism of QWZG against MASH.
METHODS
Animal models were established, including normal group, a choline-deficient, L-amino acid-defined high-fat diet (CDAHFD) group, and low/medium/high-dose QWZG groups, as well as a rosiglitazone group. Through comprehensive biochemical, histopathological, RNA sequencing, and bioinformatics analyses, galectin 3 (LGALS3) was identified as a critical target of QWZG in the treatment of MASH. The level of LGALS3 was quantitatively assessed and validated using Western blotting, real-time quantitative PCR, and immunofluorescence. The role and function of LGALS3 in inflammation and MASH progression were further investigated through gene knockdown, overexpression, iron assay, and transmission electron microscopy.
RESULTS
QWZG significantly ameliorated liver pathology by reducing steatosis, inflammation, and fibrosis. RNA sequencing analysis identified 1507 co-expressed differentially expressed genes among the CDAHFD, normal, and QWZG groups. Among these, LGALS3 was identified as one of the most significantly altered differentially expressed genes. Both mRNA and protein levels of LGALS3 were elevated in the CDAHFD group compared to the normal group, whereas treatment with QWZG reduced their levels. Analysis of Human Protein Atlas database indicated that LGALS3 was predominantly expressed in Kupffer cells, and was validated by real-time quantitative PCR and immunofluorescence. Furthermore, the level of LGALS3 was significantly increased in lipopolysaccharide-induced RAW264.7 cells, where its overexpression and recombinant LGALS3 protein both significantly enhanced the expression of interleukin-6, interleukin-1β, and tumor necrosis factor-α. LGALS3 overexpression significantly inhibited glutathione peroxidase 4 (GPX4) expression, and exacerbated mitochondrial damage, whereas LGALS3 knockdown markedly increased GPX4 level, and significantly reduced the levels of both total iron and ferrous iron. QWZG treatment significantly reduced the levels of malondialdehyde and ferrous iron, increased the levels of superoxide dismutase and glutathione. In addition, QWZG treatment also significantly enhanced GPX4 expression. Mechanistically, LGALS3 knockdown was associated with reduced expression of tumor necrosis factor receptor-associated factor 6 (TRAF6) and NOD-like receptor family pyrin domain containing 3, while its overexpression led to increased levels of these proteins. The TRAF6 inhibitor C25-140 effectively reversed the LGALS3-induced alterations in GPX4 expression and iron accumulation. Furthermore, QWZG treatment significantly decreased the levels of TRAF6 and NOD-like receptor family pyrin domain containing 3.
CONCLUSION
QWZG ameliorated the progression of MASH by modulating ferroptosis through the LGALS3/TRAF6/GPX4 axis.
Core Tip: This study reveals that galectin 3 plays a critical role in the progression of metabolic dysfunction-associated steatohepatitis by regulating ferroptosis through the tumor necrosis factor receptor-associated factor 6/glutathione peroxidase 4 axis. The traditional Chinese medicine Qiweizhigan granule significantly alleviates the progression of metabolic dysfunction-associated steatohepatitis by targeting the galectin 3/tumor necrosis factor receptor-associated factor 6/glutathione peroxidase 4 axis.
