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. 2019 Aug 12;9:11636. doi: 10.1038/s41598-019-48213-1

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Vitamin E in RWPE-1 cell line induces CYP gene expression coupled with increased ROS release and inflammation markers. (a) Gene expression study on RWPE-1 cells revealed sharp increments in mRNA levels from vitamin E-exposed cells compared to control. CYP1A1 and CYP1A4 are doubled (P < 0.05; n = 3). Isoforms 2B6, 2C9 and 4F2 showed the most pronounced changes with 18, 4 and 7 fold-higher levels respectively (P < 0.05; n = 3). (b) Free radical measurements through EPR spectroscopy in RWPE-1 cell line. The scattered dot plot represents total oxygen, nitrogen, and carbon-centered free radical species (arbitrary units) measured in RWPE-1 cells 48 hours after vitamin E exposure. Exposed cells (n = 5) exhibited significantly higher levels of radical species compared to controls (n = 4; P < 0.05). (c) Representative EPR spectra of nitroxide radicals observed in RWPE-1 control samples (green spectra), and in vitamin E-exposed samples (red spectra). EPR intensity of the first spectral line was used to obtain the relative amount of nitroxide in each sample examined. (d) Vitamin E-exposed cells (100 μM; 48 h) exhibited higher COX-2 mRNA levels (P < 0.05; n = 3). (e) COX-2 mRNA induction was coupled with an increased PGE2 secretion (P < 0.01; n = 3). Each bar represents the mean (±SD) P < 0.05, **P < 0.01 two-tailed t-test. (Control vs groups of each treatment).