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Melatonin and vitamin E alleviate homocysteine‐induced oxidative injury and apoptosis in endothelial cells

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Abstract

A relationship exists between hyperhomocysteinemia and cardiovascular diseases, although the underlying mechanisms are still incompletely defined. One possibility involves a homocysteine (Hcy)-induced increased oxidative stress. Melatonin (Mel) and vitamin E (vitE) are important anti-oxidants. The main purpose of this study was (1) to compare the effect of treatments with Mel, vitE or both, on Hcy-induced apoptosis in human umbilical vein endothelial cells (HUVECs), and (2) to investigate the underlying mechanisms. Cell proliferation assay was carried out by Water Soluble Tetrazolium-1 (WST-1) assay kit. Apoptotic index was calculated by TUNEL Assay. Anti-oxidant parameters were studied by measurement of reactive oxygen species (ROS) and lipid peroxidation (LPO) levels. mRNA and protein expression levels of apoptotic and anti-apoptotic genes and proteins were studied by quantitative real time polymerase chain reaction (qRT-PCR) and Western blotting experiments respectively. The results showed that treatments with Mel, vitE or Mel + vitE suppressed Hcy-induced cell death, with a higher efficiency for the Mel and Mel + vitE treatments. Our results suggests that the mechanisms by which these anti-oxidants protected endothelial cells include the decrease in ROS and LPO levels, an increase in cell migration, the downregulation of pro-apoptotic proteins Cas 3, Cas 9, Cyt C and Bax and the upregulation of anti-apoptotic protein Bcl 2. Collectively, these results revealed the protective role of vitE and Mel against Hcy-induced cell apoptosis, which may add insight into therapeutic approaches to Hcy-induced damages.

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Acknowledgements

This work was supported by a grant from Bingol University Independent Research Projects Office (BÜBAP), (Turkey). (Grant Number: BAP-261-278-2015). We would like to thank the directorate of Central Research Laboratory of the Bingol University for allowing us to use their laboratory facilities.

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Authors and Affiliations

  1. Department of Molecular Biology and Genetics, Faculty of Arts and Sciences, Bingol University, Bingol, Turkey

    Gurkan Aykutoglu, Musa Tartik & Ekrem Darendelioglu

  2. Department of Chemistry, Faculty of Arts and Sciences, Bingol University, Bingol, Turkey

    Adnan Ayna

  3. Department of Physiology, Faculty of Medicine, Altınbas University, Istanbul, Turkey

    Giyasettin Baydas

  4. Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden

    Musa Tartik

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  1. Gurkan Aykutoglu
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  2. Musa Tartik
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  3. Ekrem Darendelioglu
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  4. Adnan Ayna
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  5. Giyasettin Baydas
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Contributions

ED, MT and GB conceived and designed research. GA, MT and ED conducted experiments. GA, ED, MT contributed new reagents or analytical tools. MT, AA, ED and GB analysed data. AA, MT, ED and GB wrote the manuscript. The authors read and approved the manuscript.

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Correspondence to Ekrem Darendelioglu.

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Aykutoglu, G., Tartik, M., Darendelioglu, E. et al. Melatonin and vitamin E alleviate homocysteine‐induced oxidative injury and apoptosis in endothelial cells. Mol Biol Rep 47, 5285–5293 (2020). https://doi.org/10.1007/s11033-020-05607-z

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