Flavin Oxidase-Induced ROS Generation Modulates PKC Biphasic Effect of Resveratrol on Endothelial Cell Survival

<p dir="ltr">Background: Dietary intake of natural antioxidants is thought to impart protection against oxidative-associated cardiovascular diseases. Despite many in vivo studies and clinical trials, this issue has not been conclusively resolved. Resveratrol (RES) is one of the most...

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التفاصيل البيبلوغرافية
المؤلف الرئيسي: Anna Maria Posadino (13917535) (author)
مؤلفون آخرون: Roberta Giordo (299595) (author), Annalisa Cossu (299593) (author), Gheyath K. Nasrallah (9200525) (author), Abdullah Shaito (17746962) (author), Haissam Abou-Saleh (497840) (author), Ali H. Eid (5461829) (author), Gianfranco Pintus (91638) (author)
منشور في: 2019
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author Anna Maria Posadino (13917535)
author2 Roberta Giordo (299595)
Annalisa Cossu (299593)
Gheyath K. Nasrallah (9200525)
Abdullah Shaito (17746962)
Haissam Abou-Saleh (497840)
Ali H. Eid (5461829)
Gianfranco Pintus (91638)
author2_role author
author
author
author
author
author
author
author_facet Anna Maria Posadino (13917535)
Roberta Giordo (299595)
Annalisa Cossu (299593)
Gheyath K. Nasrallah (9200525)
Abdullah Shaito (17746962)
Haissam Abou-Saleh (497840)
Ali H. Eid (5461829)
Gianfranco Pintus (91638)
author_role author
dc.creator.none.fl_str_mv Anna Maria Posadino (13917535)
Roberta Giordo (299595)
Annalisa Cossu (299593)
Gheyath K. Nasrallah (9200525)
Abdullah Shaito (17746962)
Haissam Abou-Saleh (497840)
Ali H. Eid (5461829)
Gianfranco Pintus (91638)
dc.date.none.fl_str_mv 2019-05-30T03:00:00Z
dc.identifier.none.fl_str_mv 10.3390/biom9060209
dc.relation.none.fl_str_mv https://figshare.com/articles/journal_contribution/Flavin_Oxidase-Induced_ROS_Generation_Modulates_PKC_Biphasic_Effect_of_Resveratrol_on_Endothelial_Cell_Survival/25348315
dc.rights.none.fl_str_mv CC BY 4.0
info:eu-repo/semantics/openAccess
dc.subject.none.fl_str_mv Biological sciences
Biochemistry and cell biology
Zoology
ROS
resveratrol
endothelial cells
anti- and pro-oxidant effect
dose-dependence
cell damage
PKC
flavin oxidase
dc.title.none.fl_str_mv Flavin Oxidase-Induced ROS Generation Modulates PKC Biphasic Effect of Resveratrol on Endothelial Cell Survival
dc.type.none.fl_str_mv Text
Journal contribution
info:eu-repo/semantics/publishedVersion
text
contribution to journal
description <p dir="ltr">Background: Dietary intake of natural antioxidants is thought to impart protection against oxidative-associated cardiovascular diseases. Despite many in vivo studies and clinical trials, this issue has not been conclusively resolved. Resveratrol (RES) is one of the most extensively studied dietary polyphenolic antioxidants. Paradoxically, we have previously demonstrated that high RES concentrations exert a pro-oxidant effect eventually elevating ROS levels leading to cell death. Here, we further elucidate the molecular determinants underpinning RES-induced oxidative cell death. Methods: Using human umbilical vein endothelial cells (HUVECs), the effect of increasing concentrations of RES on DNA synthesis and apoptosis was studied. In addition, mRNA and protein levels of cell survival or apoptosis genes, as well as protein kinase C (PKC) activity were determined. Results: While high concentrations of RES reduce PKC activity, inhibit DNA synthesis and induce apoptosis, low RES concentrations elicit an opposite effect. This biphasic concentration-dependent effect (BCDE) of RES on PKC activity is mirrored at the molecular level. Indeed, high RES concentrations upregulate the proapoptotic Bax, while downregulating the antiapoptotic Bcl-2, at both mRNA and protein levels. Similarly, high RES concentrations downregulate the cell cycle progression genes, c-myc, ornithine decarboxylase (ODC) and cyclin D1 protein levels, while low RES concentrations display an increasing trend. The BCDE of RES on PKC activity is abrogated by the ROS scavenger Tempol, indicating that this enzyme acts downstream of the RES-elicited ROS signaling. The RES-induced BCDE on HUVEC cell cycle machinery was also blunted by the flavin inhibitor diphenyleneiodonium (DPI), implicating flavin oxidase-generated ROS as the mechanistic link in the cellular response to different RES concentrations. Finally, PKC inhibition abrogates the BCDE elicited by RES on both cell cycle progression and pro-apoptotic gene expression in HUVECs, mechanistically implicating PKC in the cellular response to different RES concentrations. Conclusions: Our results provide new molecular insight into the impact of RES on endothelial function/dysfunction, further confirming that obtaining an optimal benefit of RES is concentration-dependent. Importantly, the BCDE of RES could explain why other studies failed to establish the cardio-protective effects mediated by natural antioxidants, thus providing a guide for future investigation looking at cardio-protection by natural antioxidants.</p><p dir="ltr"><br></p><h2>Other Information</h2><p dir="ltr">Published in: Biomolecules<br>License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.3390/biom9060209" target="_blank">https://dx.doi.org/10.3390/biom9060209</a></p>
eu_rights_str_mv openAccess
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identifier_str_mv 10.3390/biom9060209
network_acronym_str Manara2
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oai_identifier_str oai:figshare.com:article/25348315
publishDate 2019
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spelling Flavin Oxidase-Induced ROS Generation Modulates PKC Biphasic Effect of Resveratrol on Endothelial Cell SurvivalAnna Maria Posadino (13917535)Roberta Giordo (299595)Annalisa Cossu (299593)Gheyath K. Nasrallah (9200525)Abdullah Shaito (17746962)Haissam Abou-Saleh (497840)Ali H. Eid (5461829)Gianfranco Pintus (91638)Biological sciencesBiochemistry and cell biologyZoologyROSresveratrolendothelial cellsanti- and pro-oxidant effectdose-dependencecell damagePKCflavin oxidase<p dir="ltr">Background: Dietary intake of natural antioxidants is thought to impart protection against oxidative-associated cardiovascular diseases. Despite many in vivo studies and clinical trials, this issue has not been conclusively resolved. Resveratrol (RES) is one of the most extensively studied dietary polyphenolic antioxidants. Paradoxically, we have previously demonstrated that high RES concentrations exert a pro-oxidant effect eventually elevating ROS levels leading to cell death. Here, we further elucidate the molecular determinants underpinning RES-induced oxidative cell death. Methods: Using human umbilical vein endothelial cells (HUVECs), the effect of increasing concentrations of RES on DNA synthesis and apoptosis was studied. In addition, mRNA and protein levels of cell survival or apoptosis genes, as well as protein kinase C (PKC) activity were determined. Results: While high concentrations of RES reduce PKC activity, inhibit DNA synthesis and induce apoptosis, low RES concentrations elicit an opposite effect. This biphasic concentration-dependent effect (BCDE) of RES on PKC activity is mirrored at the molecular level. Indeed, high RES concentrations upregulate the proapoptotic Bax, while downregulating the antiapoptotic Bcl-2, at both mRNA and protein levels. Similarly, high RES concentrations downregulate the cell cycle progression genes, c-myc, ornithine decarboxylase (ODC) and cyclin D1 protein levels, while low RES concentrations display an increasing trend. The BCDE of RES on PKC activity is abrogated by the ROS scavenger Tempol, indicating that this enzyme acts downstream of the RES-elicited ROS signaling. The RES-induced BCDE on HUVEC cell cycle machinery was also blunted by the flavin inhibitor diphenyleneiodonium (DPI), implicating flavin oxidase-generated ROS as the mechanistic link in the cellular response to different RES concentrations. Finally, PKC inhibition abrogates the BCDE elicited by RES on both cell cycle progression and pro-apoptotic gene expression in HUVECs, mechanistically implicating PKC in the cellular response to different RES concentrations. Conclusions: Our results provide new molecular insight into the impact of RES on endothelial function/dysfunction, further confirming that obtaining an optimal benefit of RES is concentration-dependent. Importantly, the BCDE of RES could explain why other studies failed to establish the cardio-protective effects mediated by natural antioxidants, thus providing a guide for future investigation looking at cardio-protection by natural antioxidants.</p><p dir="ltr"><br></p><h2>Other Information</h2><p dir="ltr">Published in: Biomolecules<br>License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.3390/biom9060209" target="_blank">https://dx.doi.org/10.3390/biom9060209</a></p>2019-05-30T03:00:00ZTextJournal contributioninfo:eu-repo/semantics/publishedVersiontextcontribution to journal10.3390/biom9060209https://figshare.com/articles/journal_contribution/Flavin_Oxidase-Induced_ROS_Generation_Modulates_PKC_Biphasic_Effect_of_Resveratrol_on_Endothelial_Cell_Survival/25348315CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/253483152019-05-30T03:00:00Z
spellingShingle Flavin Oxidase-Induced ROS Generation Modulates PKC Biphasic Effect of Resveratrol on Endothelial Cell Survival
Anna Maria Posadino (13917535)
Biological sciences
Biochemistry and cell biology
Zoology
ROS
resveratrol
endothelial cells
anti- and pro-oxidant effect
dose-dependence
cell damage
PKC
flavin oxidase
status_str publishedVersion
title Flavin Oxidase-Induced ROS Generation Modulates PKC Biphasic Effect of Resveratrol on Endothelial Cell Survival
title_full Flavin Oxidase-Induced ROS Generation Modulates PKC Biphasic Effect of Resveratrol on Endothelial Cell Survival
title_fullStr Flavin Oxidase-Induced ROS Generation Modulates PKC Biphasic Effect of Resveratrol on Endothelial Cell Survival
title_full_unstemmed Flavin Oxidase-Induced ROS Generation Modulates PKC Biphasic Effect of Resveratrol on Endothelial Cell Survival
title_short Flavin Oxidase-Induced ROS Generation Modulates PKC Biphasic Effect of Resveratrol on Endothelial Cell Survival
title_sort Flavin Oxidase-Induced ROS Generation Modulates PKC Biphasic Effect of Resveratrol on Endothelial Cell Survival
topic Biological sciences
Biochemistry and cell biology
Zoology
ROS
resveratrol
endothelial cells
anti- and pro-oxidant effect
dose-dependence
cell damage
PKC
flavin oxidase