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Protection by chrysin, apigenin, and luteolin against oxidative stress is mediated by the Nrf2-dependent up-regulation of heme oxygenase 1 and glutamate cysteine ligase in rat primary hepatocytes

  • Organ Toxicity and Mechanisms
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Abstract

Chrysin, apigenin, and luteolin are flavones that differ in their number of hydroxyl groups in the B ring. In this study, we investigated the protection by chrysin, apigenin, and luteolin against tert-butyl hydroperoxide (tBHP)-induced oxidative stress and the possible mechanisms involved in rat primary hepatocytes. Chrysin, apigenin, and luteolin dose-dependently up-regulated the protein expression of heme oxygenase 1 (HO-1) and glutamate cysteine ligase (GCL) catalytic (GCLC) and modifier subunit (GCLM) and increased the intracellular glutathione (GSH) content and the ratio of GSH to oxidized GSH. Among the flavones studied, chrysin showed the greatest induction of HO-1, GCLC, and GCLM protein expression and GSH content. Cellular reactive oxygen species production induced by tBHP was attenuated by pretreatment with chrysin, apigenin, and luteolin (P < .05), and this protection was reversed by the GCL inhibitor l-buthionine-S-sulfoximine and the HO-1 inhibitor zinc protoporphyrin. Chrysin, apigenin, and luteolin activated extracellular signal-regulated protein kinase 2 (ERK2), nuclear factor erythroid 2-related factor 2 (Nrf2) nuclear translocation, nuclear Nrf2–antioxidant responsive element (ARE) binding activity, and ARE-dependent luciferase activity. Both ERK2 and Nrf2 siRNAs attenuated chrysin-induced HO-1, GCLC, and GCLM protein expression. Taken together, these results suggest that chrysin, apigenin, and luteolin inhibit tBHP-induced oxidative stress by up-regulating HO-1, GCLC, and GCLM gene transcription via the ERK2/Nrf2/ARE signaling pathways in rat primary hepatocytes.

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Acknowledgments

This study was supported by grant CMU99-ASIA-29.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Health and Nutrition Biotechnology, Asia University, Taichung, Taiwan, ROC

    Chin-Shiu Huang

  2. Department of Nutrition, China Medical University, Taichung, Taiwan, ROC

    Chong-Kuei Lii, Yu-Wen Yeh, Hsien-Tsung Yao & Haw-Wen Chen

  3. Department of Nutrition, Chung Shan Medical University, Taichung, Taiwan, ROC

    Ai-Hsuan Lin & Chien-Chun Li

  4. Department of Biomedical Sciences, Chung Shan Medical University, Taichung, Taiwan, ROC

    Tsu-Shing Wang

  5. Department of Medical Research, Chung Shan Medical University Hospital, Taichung, Taiwan, ROC

    Tsu-Shing Wang

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  1. Chin-Shiu Huang
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  5. Hsien-Tsung Yao
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Correspondence to Tsu-Shing Wang or Haw-Wen Chen.

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Chin-Shiu Huang and Chong-Kuei Lii have contributed equally to this work.

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Huang, CS., Lii, CK., Lin, AH. et al. Protection by chrysin, apigenin, and luteolin against oxidative stress is mediated by the Nrf2-dependent up-regulation of heme oxygenase 1 and glutamate cysteine ligase in rat primary hepatocytes. Arch Toxicol 87, 167–178 (2013). https://doi.org/10.1007/s00204-012-0913-4

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