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Neoxanthin prevents H2O2-induced cytotoxicity in HepG2 cells by activating endogenous antioxidant signals and suppressing apoptosis signals

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Abstract

Background

The liver has a solid inbuilt antioxidant defense system to regulate oxidative stress. However, exposure to an excessive level of ROS causes liver injury. This study examined the cytoprotective effect of neoxanthin, a xanthophyll antioxidant molecule isolated from Solanum trilobatum in stress-induced HepG2 cells.

Methods and results

The cytotoxic effect of H2O2 and cytoprotective potential of β-carotene, lutein, and neoxanthin was analyzed by WST-1 assay. The intracellular ROS level and mitochondrial membrane potential (MMP) were measured using DCFH-DA (2′, 7′-dichlorofluorescin diacetate) and JC-10 MMP assay. The expression of anti-oxidant and apoptotic markers was measured by western blot analysis. Neoxanthin pretreatment exhibited better protection than β-carotene and lutein against cell death caused by H2O2. It significantly arrested H2O2-mediated elevation of intracellular ROS levels and protected MMP. The intracellular antioxidant enzymes HO-1 and SOD-2 were upregulated by neoxanthin pretreatment. Neoxanthin also activated the protein expression of redox-sensitive transactivation factors, Nrf2 and NF-kB. The cytoprotective effect of neoxanthin was associated with increased expression of the anti-apoptotic protein, Bcl-2 and decreased pro-apoptotic protein Bax.

Conclusions

For the first time, our results demonstrate that neoxanthin offers adequate protection against stress-mediated cytotoxicity in hepatocytes by activating the intracellular antioxidant defense system and blocking apoptosis.

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Acknowledgements

The author, TM, acknowledges the Department of Biotechnology (DBT), New Delhi, India, for granting a research fellowship. The authors thank the Director, CSIR-CFTRI, for the constant support to carry out this work.

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Author notes

  1. Deepika Udayawara Rudresh and Tehreem Maradagi have contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Nutrition, CSIR-Central Food Technological Research Institute (CFTRI), Mysuru, 570 020, India

    Deepika Udayawara Rudresh, Tehreem Maradagi & Ganesan Ponesakki

  2. Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201 002, India

    Tehreem Maradagi, Ayyadurai Niraikulam, Kamini Nambi Ramudu & Ganesan Ponesakki

  3. Department of Fish Processing Technology, Dr. MGR Fisheries College and Research Institute, Tamil Nadu Dr. J. Jeyalalithaa Fisheries University, Ponneri, 601 204, India

    Nimish Mol Stephen

  4. Department of Biochemistry and Biotechnology, CSIR-Central Leather Research Institute (CLRI), Adyar, Chennai, 600 020, India

    Ayyadurai Niraikulam, Kamini Nambi Ramudu & Ganesan Ponesakki

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  1. Deepika Udayawara Rudresh
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Contributions

GP, DUR, NMS and TM designed the study; DUR, TM and NMS performed the experiments; GP, DUR, TM and NMS wrote the manuscript; AN and KNR contributed to data analysis and editing of the manuscript, and all the authors approved the final version of the manuscript.

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Correspondence to Ganesan Ponesakki.

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Udayawara Rudresh, D., Maradagi, T., Stephen, N.M. et al. Neoxanthin prevents H2O2-induced cytotoxicity in HepG2 cells by activating endogenous antioxidant signals and suppressing apoptosis signals. Mol Biol Rep 48, 6923–6934 (2021). https://doi.org/10.1007/s11033-021-06695-1

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