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Sulfuretin protects hepatic cells through regulation of ROS levels and autophagic flux

Acta Pharmacologica Sinica (2018) | Download Citation

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Abstract

Palmitate (PA) exposure induces stress conditions featuring ROS accumulation and upregulation of p62 expression, resulting in autophagic flux blockage and cell apoptosis. Sulfuretin (Sul) is a natural product isolated from Rhus verniciflua Stokes; the cytoprotective effect of Sul on human hepatic L02 cells and mouse primary hepatocytes under PA-induced stress conditions was investigated in this study. Sul induced mitophagy by activation of p-TBK1 and LC3 and produced a concomitant decline in p62 expression. Autophagosome formation and mitophagy were assessed by the sensitive dual fluorescence reporter mCherry-EGFP-LC3B, and mitochondrial fragmentation was analyzed using MitoTracker Deep Red FM. A preliminary structure–activity relationship (SAR) for Sul was also investigated, and the phenolic hydroxyl group was found to be pivotal for maintaining the cytoprotective bioactivity of Sul. Furthermore, experiments using flow cytometry and western blots revealed that Sul reversed the cytotoxic effect stimulated by the autophagy inhibitors 3-methyladenine (3-MA) and chloroquine (CQ), and its cytoprotective effect was almost eliminated when the autophagy-related 5 (Atg5) gene was knocked down. These studies suggest that, in addition to its antioxidative effects, Sul stimulates mitophagy and restores impaired autophagic flux, thus protecting hepatic cells from apoptosis, and that Sul has potential future medical applications for hepatoprotection.

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Acknowledgements

This work was supported by a grant from the Shanghai Commission of Science and Technology (16JC1405000) and K.C. Wong Education Foundation.

Author contributions

Y.-T.L. and Y.-F.L. conducted the biological experiments, analyzed the results and wrote the paper; Y.-F.X. conducted the chemical experiments, analyzed the results and wrote the paper; J.-Y.L., F.-J.N., and J.L. conceived the idea for the project, analyzed the results and wrote the paper. All authors reviewed the results and approved the final version of the manuscript.

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Author notes

  1. These authors contributed equally: Yu-Ting Lu, Yu-Feng Xiao.

Affiliations

  1. State Key Laboratory of Drug Research, The National Center for Drug Screening, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 201203, Shanghai, China

    • Yu-Ting Lu
    • , Yu-Feng Xiao
    • , Yu-Feng Li
    • , Jia Li
    • , Fa-Jun Nan
    •  & Jing-Ya Li
  2. University of Chinese Academy of Sciences, 100049, Beijing, China

    • Yu-Ting Lu
    • , Yu-Feng Xiao
    • , Yu-Feng Li
    • , Jia Li
    • , Fa-Jun Nan
    •  & Jing-Ya Li
  3. School of Life Science and Technology, ShanghaiTech University, 201210, Shanghai, China

    • Yu-Ting Lu
    •  & Jia Li

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The authors declare that they have no conflict of interest.

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Correspondence to Jia Li or Fa-Jun Nan or Jing-Ya Li.

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DOI

https://doi.org/10.1038/s41401-018-0193-5