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Self-protection against triptolide-induced toxicity in human hepatic cells via Nrf2-ARE-NQO1 pathway

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Abstract

Objective

To find the signaling pathway of triptolide (TP)-induced liver injury and to reveal whether NF-E2-related factor 2 (Nrf2) plays an important role in cellular self-protection.

Methods

The L-02 and HepG2 cells were cultured and treated with various concentrations of TP. The cell viability was observed, and the cell medium was collected for detecting the aspartate aminotransferase (ALT), alanine aminotransferase (AST), lactate dehydrogenase (LDH), superoxide dismutase (SOD) and L-glutathione production (GSH) levels. Nrf2 and its downstream target NAD(P)H: quinine oxidoreductase 1 (NQO1) and heme oxygenase-1 (HO-1) expression, the nuclear translocation of Nrf2, and the binding ability of Nrf2 and antioxidant response element (ARE) were also identified. Meanwhile, shRNA was used to silence Nrf2 in L-02 cells to find out whether Nrf2 plays a protective role.

Results

The viability of the L-02 and HepG2 cells treated with TP decreased in a doseand time-dependent manner, and TP (20–80 μg/mL) markedly induced the release of ALT, AST and LDH (P<0.05 or P<0.01), reduced the levels of SOD and GSH (P<0.01), and increased the intracellular reactive oxygen species. Meanwhile, TP augmented the Nrf2 expression in L-02 and HepG2 cells (P<0.05 or P<0.01), induced Nrf2 nuclear translocation, increased the Nrf2 ARE binding activity, and increased HO-1 and NQO1 expressions. Nrf2 knockdown revealed a more severe toxic effect of TP (P<0.05 or P<0.01).

Conclusions

Human hepatic cells treated with TP induced oxidative stress, and led to cytotoxicity. Self-protection against TP-induced toxicity in human hepatic cells might be via Nrf2-ARE-NQO1 transcriptional pathway.

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Authors and Affiliations

  1. School of Pharmacy, The First Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing, 210023, China

    Ling-ling Zhou, Cong Zhou, Zhe Feng, Zhang-pu Liu & Xue-ping Zhou

  2. Department of Pharmacy, Wuxi No.9 People’s Hospital, Soochow University, Wuxi Hand Surgery Hospital, Wuxi, Jiangsu Province, 214062, China

    Cong Zhou

  3. Therapeutics Research Centre, School of Medicine, The University of Queensland, Princess Alexandra Hospital, 4102, Woolloongabba, QLD, Australia

    Xiao-wen Liang & Hao-lu Wang

Authors
  1. Ling-ling Zhou
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  2. Cong Zhou
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  3. Xiao-wen Liang
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  4. Zhe Feng
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  5. Zhang-pu Liu
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  6. Hao-lu Wang
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  7. Xue-ping Zhou
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Correspondence to Xue-ping Zhou.

Additional information

Supported by National Natural Science Foundation of China (No. 81072749, 81573869), National Natural Science Foundation for the Youth of Jiangsu Province (No. BK20140960) and National Natural Science Pre-research of Nanjing University of Chinese Medicine (No. 14XYY01, 14XYY10)

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Zhou, Ll., Zhou, C., Liang, Xw. et al. Self-protection against triptolide-induced toxicity in human hepatic cells via Nrf2-ARE-NQO1 pathway. Chin. J. Integr. Med. 23, 929–936 (2017). https://doi.org/10.1007/s11655-017-2546-6

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  • DOI: https://doi.org/10.1007/s11655-017-2546-6

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