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Targeting glutathione with the triterpenoid CDDO-Im protects against benzo-a-pyrene-1,6-quinone-induced cytotoxicity in endothelial cells

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Abstract

Epidemiological studies have exhibited a strong correlation between exposure to air pollution and deaths due to vascular diseases such as atherosclerosis. Benzo-a-pyrene-1,6-quinone (BP-1,6-Q) is one of the components of air pollution. This study was to examine the role of GSH in BP-1,6-Q mediated cytotoxicity in human EA.hy96 endothelial cells and demonstrated that induction of cellular glutathione by a potent triterpenoid, CDDO-Im (1-[2-cyano-3-,12-dioxooleana-1,9(11)-dien-28-oyl]imidazole), protects cells against BP-1,6-Q induced protein and lipid damage. Incubation of EA.hy926 endothelial cells with BP-1,6-Q caused a significant increase in dose-dependent cytotoxicity as measured by LDH release assay and both apoptotic and necrotic cell deaths as measured by flow cytometric analysis. Incubation of EA.hy926 endothelial cells with BP-1,6-Q also caused a significant decrease in cellular GSH levels. The diminishment of cellular GSH by buthionine sulfoximine (BSO) potentiated BP-1,6-Q-induced toxicity significantly suggesting a critical involvement of GSH in BP-1,6-Q induced cellular toxicity. GSH-induction by CDDO-Im significantly protects cells against BP-1,6-Q induced protein and lipid damage as measured by protein carbonyl (PC) assay and thiobarbituric acid reactive substances (TBARS) assay, respectively. However, the co-treatment of cells with CDDO-Im and BSO reversed the cytoprotective effect of CDDO-Im on BP-1,6-Q-mediated lipid peroxidation and protein oxidation. These results suggest that induction of GSH by CDDO-Im might be the important cellular defense against BP-1,6-Q induced protein and lipid damage. These findings would contribute to better understand the action of BP-1,6-Q and may help to develop novel therapies to protect against BP-1,6-Q-induced atherogenesis.

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Data availability

The data of this study can be obtained from the corresponding author upon reasonable request.

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Acknowledgements

This work was supported by a grant from UNCG.

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Authors and Affiliations

  1. Department of Biology, The University of North Carolina At Greensboro, 312 Eberhart Building, 321 McIver Street, Greensboro, NC, 27402-6170, USA

    Halley Shukla, Ho Young Lee, Ashkon Koucheki, Humaira A. Bibi, Gabriella Gaje & Zhenquan Jia

  2. Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, AR, 72701, USA

    Xiaolun Sun

  3. Campbell University School of Osteopathic Medicine, Buies Creek, NC, USA

    Hong Zhu & Y. Robert Li

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  1. Halley Shukla
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  2. Ho Young Lee
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Contributions

ZJ designed the experiments. HS, ZJ, HYL, AK, HAB, GG completed the experiments and processed the experimental data analysis with substantial input provided by YRL, HZ, and XS. YRL, HZ, and XS also provided useful comments and feedback for the manuscript.

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Correspondence to Zhenquan Jia.

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Shukla, H., Lee, H.Y., Koucheki, A. et al. Targeting glutathione with the triterpenoid CDDO-Im protects against benzo-a-pyrene-1,6-quinone-induced cytotoxicity in endothelial cells. Mol Cell Biochem 474, 27–39 (2020). https://doi.org/10.1007/s11010-020-03831-6

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