Copyright
©The Author(s) 2026. Published by Baishideng Publishing Group Inc. All rights reserved.
Qushi Huoxue ointment ameliorates metabolic associated steatotic liver disease through autophagy activation and ferroptosis inhibition
Yi-Yang Liu, Hong Qin, Hong-Xi Wu, Ru-Ting Wang, Qiu-Yan Yang, Feng Jiang, Xu-Dong Liu, De-Kun Wu, You-Ming Tang
Yi-Yang Liu, Hong Qin, Hong-Xi Wu, Ru-Ting Wang, Qiu-Yan Yang, Graduate School, Guangxi University of Chinese Medicine, Nanning 530011, Guangxi Zhuang Autonomous Region, China
Feng Jiang, Medical Translation Center, Ruikang Hospital Affiliated to Guangxi University of Traditional Chinese Medicine, Nanning 530011, Guangxi Zhuang Autonomous Region, China
Xu-Dong Liu, Department of Hepatology, Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine, Nanning 530011, Guangxi Zhuang Autonomous Region, China
De-Kun Wu, Teaching Experiment and Training Center, Guangxi University of Chinese Medicine, Nanning 530011, Guangxi Zhuang Autonomous Region, China
You-Ming Tang, Department of Digestive Disease, Ruikang Hospital Affiliated to Guangxi University of Traditional Chinese Medicine, Nanning 530011, Guangxi Zhuang Autonomous Region, China
Co-corresponding authors: De-Kun Wu and You-Ming Tang.
Author contributions: Liu YY designed the experimental protocol, conducted the majority of the animal and molecular biology experiments, analyzed the corresponding data, and drafted the paper; Qin H was responsible for data processing, statistical analysis and chart creation, and assisted in revising the paper; Wu HX, Wang RT, and Yang QY assisted in conducting western blot and qPCR experiments and participated in the preliminary data analysis; Jiang F and Liu XD provided key experimental reagents and technical guidance, supervised the research progress, and critically reviewed and revised the paper; Wu DK and Tang YM secured research funding, established the overall research framework, finalized the manuscript, and they contributed to the work equally to this article and are co-corresponding authors. All authors contributed to the study conception and design, have read and approved the final version of the manuscript.
Supported by the National Natural Science Foundation of China, No. 82160837; Huatong Guokang Medical Research Special Project Grant, No. 2023HT026; and Innovation Project of Guangxi Graduate Education, No. YCSW2024405.
Institutional animal care and use committee statement: This study was reviewed and approved by the Animal Welfare and Ethics Committee of Guangxi University of Chinese Medicine (Approval No. DW20240919-186).
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
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: No additional data are available.
Open Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
https://creativecommons.org/Licenses/by-nc/4.0/ Corresponding author: De-Kun Wu, Teaching Experiment and Training Center, Guangxi University of Chinese Medicine, No. 179 Mingxiu East Road, Xixiangtang District, Nanning 530011, Guangxi Zhuang Autonomous Region, China.
1278323777@qq.com
Received: November 3, 2025
Revised: November 23, 2025
Accepted: December 25, 2025
Published online: February 27, 2026
Processing time: 104 Days and 5.1 Hours
BACKGROUND
Metabolic associated steatotic liver disease (MASLD) has become a growing global health burden, with its potential to progress to liver fibrosis, cirrhosis, and hepatocellular carcinoma. Qushi Huoxue ointment (QSHXO), a traditional Chinese medicine formula, has demonstrated efficacy in the management of MASLD. However, its underlying mechanisms remain incompletely elucidated.
AIM
To investigate the mechanism by which QSHXO alleviated hepatic lipid deposition and inflammatory injury in MASLD, with a focus on its role in activating hepatocyte autophagy and inhibiting ferroptosis.
METHODS
This study employed a comprehensive research strategy. First, a methionine-choline-deficient diet-triggered MASLD mouse model was established and treated with different doses of QSHXO. The therapeutic effects of QSHXO were comprehensively evaluated using histological analysis, serum biochemical assays, and inflammatory cytokine measurements. Subsequently, bioactive components of QSHXO in serum were identified utilizing liquid chromatography-tandem mass spectrometry. Network pharmacology was then applied to predict potential targets of QSHXO in treating MASLD related to autophagy and ferroptosis. These predicted targets were validated through western blotting, quantitative reverse-transcription polymerase chain reaction, immunohistochemistry, and transmission electron microscopy.
RESULTS
QSHXO significantly ameliorated liver lipid deposition and inflammation in MASLD mice. Specifically, QSHXO promoted autophagic flux, as indicated by upregulation of Beclin1, an increased light chain 3 II/light chain 3 I ratio, and downregulation of P62. Concurrently, QSHXO activated the nuclear factor erythroid 2-related factor 2 pathway, promoting its nuclear translocation and enhancing the expression of downstream targets (SLC7A11 and glutathione peroxidase 4), while reducing hepatic iron deposition; these collectively suggested suppression of ferroptosis. Ultrastructural analysis further confirmed improved mitochondrial morphology and increased autophagic vesicles in QSHXO-treated groups.
CONCLUSION
QSHXO ameliorates MASLD by reducing lipid accumulation, mitigating inflammation, and suppressing hepatocyte damage, which is mediated through the activation of autophagy and inhibition of ferroptosis.
Core Tip: Qushi Huoxue ointment can alleviate hepatic lipid accumulation, inflammation, and cell injury in metabolic associated steatotic liver disease mice. Our experimental evidence suggests that these therapeutic effects may be attributed to the concurrent activation of autophagy and inhibition of ferroptosis. The improvement in mitochondrial morphology and the presence of autophagosomes observed under the microscope provide morphological corroboration for this coordinated mechanism.
