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Induction of Nrf2-mediated phase II detoxifying/antioxidant enzymes in vitro by chitosan-caffeic acid against hydrogen peroxide-induced hepatotoxicity through JNK/ERK pathway

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Abstract

Chemical modification of chitosan is a promising method for the improvement of biological activity. In this study, chitosan-caffeic acid (CCA) was prepared and its in vitro hepatoprotective ability against hydrogen peroxide-induced hepatic damage in liver cells was evaluated. Treatment with CCA (50–400 µg/mL) did not show cytotoxicity and also significantly (p < 0.05) recovered cell viability against 650 µM hydrogen peroxide-induced hepatotoxicity. CCA treatment attenuated reactive oxygen species generation and lipid peroxidation in addition to increasing cellular glutathione level in cultured hepatocytes. To validate the underlying mechanism, antioxidant and phase II detoxifying enzyme expressions, which are mediated by NF-E2-related factor 2 (Nrf2) activation, were analyzed and CCA treatment was found to increase the expression of superoxide dismutase-1 (SOD-1), glutathione reductase (GR), heme oxygenase-1 (HO-1), and NAD(P)H:quinine oxidoreductase 1 (NQO1). CCA treatment resulted in increased Nrf2 nuclear translocation. The phosphorylation of extracellular regulated kinase (ERK), c-Jun NH2-terminal kinase (JNK), and p38 mitogen-activated protein kinase (MAPK) by CCA treatment contributed to Nrf2 activation. Pharmacological blockade of ERK, JNK, and p38 MAPK revealed that SP600125 (JNK inhibitor) and PD98059 (ERK inhibitor) treatment reduced Nrf2 translocation into the nucleus while SB203580 (p38 inhibitor) exhibited weak inhibition. Collectively, CCA protects liver cells against hydrogen peroxide-induced injury and this ability is attributed to the induction of antioxidants and phase II detoxifying enzymes that are mediated by Nrf2 translocation via JNK/ERK signaling.

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Abbreviations

CCA:

Chitosan-caffeic acid

SOD-1:

Superoxide dismutase-1

GR:

Glutathione reductase

HO-1:

Heme oxygenase-1

NQO1:

NAD(P)H:quinine oxidoreductase 1

Nrf2:

NF-E2-related factor 2

GSH:

Glutathione

MAPK:

Mitogen-activated protein kinase

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Acknowledgments

This work was supported by the Human Resource Training Program for Regional Innovation and Creativity through the Ministry of Education and National Research Foundation of Korea (NRF-2014H1C1A1066586).

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Authors and Affiliations

  1. Division of Food and Nutrition, Chonnam National University, Gwangju, 61186, Republic of Korea

    Chang-Bum Ahn

  2. Department of Marine-Bio Convergence Science, Pukyong National University, Busan, 48547, Republic of Korea

    Jae-Young Je

  3. Department of Chemistry, Pukyong National University, Busan, 48513, Republic of Korea

    Young-Sang Kim & Sun-Joo Park

  4. Specialized Graduate School of Science & Technology Convergence, Pukyong National University, Busan, 48547, Republic of Korea

    Boo Il Kim

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  1. Chang-Bum Ahn
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Correspondence to Jae-Young Je.

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Ahn, CB., Je, JY., Kim, YS. et al. Induction of Nrf2-mediated phase II detoxifying/antioxidant enzymes in vitro by chitosan-caffeic acid against hydrogen peroxide-induced hepatotoxicity through JNK/ERK pathway. Mol Cell Biochem 424, 79–86 (2017). https://doi.org/10.1007/s11010-016-2845-4

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  • DOI: https://doi.org/10.1007/s11010-016-2845-4

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