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Activation of the Nrf2 signaling pathway in usnic acid-induced toxicity in HepG2 cells

  • Molecular Toxicology
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Abstract

Many usnic acid-containing dietary supplements have been marketed as weight loss agents, although severe hepatotoxicity and acute liver failure have been associated with their overuse. Our previous mechanistic studies revealed that autophagy, disturbance of calcium homeostasis, and ER stress are involved in usnic acid-induced toxicity. In this study, we investigated the role of oxidative stress and the Nrf2 signaling pathway in usnic acid-induced toxicity in HepG2 cells. We found that a 24-h treatment with usnic acid caused DNA damage and S-phase cell cycle arrest in a concentration-dependent manner. Usnic acid also triggered oxidative stress as demonstrated by increased reactive oxygen species generation and glutathione depletion. Short-term treatment (6 h) with usnic acid significantly increased the protein level for Nrf2 (nuclear factor erythroid 2-related factor 2), promoted Nrf2 translocation to the nucleus, up-regulated antioxidant response element (ARE)-luciferase reporter activity, and induced the expression of Nrf2-regulated targets, including glutathione reductase, glutathione S-transferase, and NAD(P)H quinone oxidoreductase-1 (NQO1). Furthermore, knockdown of Nrf2 with shRNA potentiated usnic acid-induced DNA damage and cytotoxicity. Taken together, our results show that usnic acid causes cell cycle dysregulation, DNA damage, and oxidative stress and that the Nrf2 signaling pathway is activated in usnic acid-induced cytotoxicity.

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Acknowledgments

ZH. Z., DK. Y., and Z. R. were supported by appointments to the Postgraduate Research Program at the National Center for Toxicological Research administered by the Oak Ridge Institute for Science Education through an interagency agreement between the US Department of Energy and the U.S. FDA. This work was supported by U.S. FDA’s intramural grant. L.S. was supported by the National High Technology Research and Development Program of China (2015AA020104), the National Natural Science Foundation of China (31471239), the 111 Project (B13016), and the National Supercomputer Center in Guangzhou, China

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Authors and Affiliations

  1. Division of Biochemical Toxicology, National Center for Toxicological Research (NCTR)/U.S. Food and Drug Administration (FDA), HFT-110, 3900 NCTR Road, Jefferson, AR, 72079, USA

    Si Chen, Zhen Ren, Letha Couch, William H. Tolleson & Lei Guo

  2. Division of Genetic and Molecular Toxicology, National Center for Toxicological Research/U.S. FDA, Jefferson, AR, 72079, USA

    Zhuhong Zhang & Nan Mei

  3. Tianjin Medical University General Hospital, Tianjin, 300052, China

    Zhuhong Zhang

  4. School of Pharmacy, School of Life Sciences, Fudan-Zhangjiang Center for Clinical Genomics and Zhanjiang Center for Translational Medicine, Fudan University, Shanghai, 200438, China

    Tao Qing & Leming Shi

  5. Division of Systems Biology, National Center for Toxicological Research/U.S. FDA, Jefferson, AR, 72079, USA

    Dianke Yu & Baitang Ning

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  1. Si Chen
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  2. Zhuhong Zhang
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  10. William H. Tolleson
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Correspondence to Lei Guo.

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Chen, S., Zhang, Z., Qing, T. et al. Activation of the Nrf2 signaling pathway in usnic acid-induced toxicity in HepG2 cells. Arch Toxicol 91, 1293–1307 (2017). https://doi.org/10.1007/s00204-016-1775-y

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