Oxidative Stress Induces HSP90 Upregulation on the Surface of Primary Human Endothelial Cells: Role of the Antioxidant 7,8-Dihydroxy-4-methylcoumarin in Preventing HSP90 Exposure to the Immune System

Elisabetta Profumo, Brigitta Buttari, Lavinia Tinaburri, Daniela D'Arcangelo, Maurizio Sorice, Antonella Capozzi, Tina Garofalo, Antonio Facchiano, Rita Businaro, Prashant Kumar, Brajendra K. Singh, Virinder S. Parmar, Luciano Saso, Rachele Rigano

Research output: Contribution to journalArticle

Abstract

We have previously demonstrated that human heat shock protein 90 (HSP90), an intracellular self protein, is the target of cellular and humoral autoimmune responses in patients with carotid atherosclerosis. In this study, we evaluated in vitro whether oxidative stress, a feature of atherosclerotic plaque, alters HSP90 expression in endothelial cells, thus inducing surface localization of this molecule and whether the antioxidant compound 7,8-dihydroxy-4-methylcoumarin (7,8-DHMC) is able to prevent oxidative stress-induced alterations of HSP90 localization. By the use of flow cytometry, immunofluorescence, enzyme-linked immunosorbent assay, and semiquantitative reverse-transcription polymerase chain reaction, we demonstrated that exposure of human umbilical vein endothelial cells (HUVEC) to the prooxidant compound H2O2 upregulated HSP90 surface expression and reduced its secretion without altering HSP90 gene expression and intracytoplasmic protein levels. Pretreatment of HUVEC with 7,8-DHMC prevented H2O2-induced alterations of HSP90 cellular distribution and secretion. Our results suggest that the strong oxidative conditions of atherosclerotic plaques promote the upregulation of HSP90 surface expression on endothelial cells, thus rendering the protein a possible target of autoimmune reactions. The antioxidant 7,8-DHMC, by preventing oxidative-stress-triggered HSP90 surface upregulation, may be useful to counteract possible autoreactive reactions to HSP90.
Original languageEnglish
JournalOxidative Medicine and Cellular Longevity
Volume2018
DOIs
Publication statusPublished - Jan 1 2018

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HSP90 Heat-Shock Proteins
Oxidative stress
Immune system
Endothelial cells
Immune System
Oxidative Stress
Up-Regulation
Endothelial Cells
Antioxidants
Human Umbilical Vein Endothelial Cells
Atherosclerotic Plaques
7,8-dihydroxy-4-methylcoumarin
Immunosorbents
Proteins
Carotid Artery Diseases
Flow cytometry
Polymerase chain reaction
Transcription
Autoimmunity
Gene expression

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Oxidative Stress Induces HSP90 Upregulation on the Surface of Primary Human Endothelial Cells: Role of the Antioxidant 7,8-Dihydroxy-4-methylcoumarin in Preventing HSP90 Exposure to the Immune System. / Profumo, Elisabetta; Buttari, Brigitta; Tinaburri, Lavinia; D'Arcangelo, Daniela; Sorice, Maurizio; Capozzi, Antonella; Garofalo, Tina; Facchiano, Antonio; Businaro, Rita; Kumar, Prashant; Singh, Brajendra K.; Parmar, Virinder S.; Saso, Luciano; Rigano, Rachele.

In: Oxidative Medicine and Cellular Longevity, Vol. 2018, 01.01.2018.

Research output: Contribution to journalArticle

Profumo, Elisabetta ; Buttari, Brigitta ; Tinaburri, Lavinia ; D'Arcangelo, Daniela ; Sorice, Maurizio ; Capozzi, Antonella ; Garofalo, Tina ; Facchiano, Antonio ; Businaro, Rita ; Kumar, Prashant ; Singh, Brajendra K. ; Parmar, Virinder S. ; Saso, Luciano ; Rigano, Rachele. / Oxidative Stress Induces HSP90 Upregulation on the Surface of Primary Human Endothelial Cells: Role of the Antioxidant 7,8-Dihydroxy-4-methylcoumarin in Preventing HSP90 Exposure to the Immune System. In: Oxidative Medicine and Cellular Longevity. 2018 ; Vol. 2018.
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AU - Buttari, Brigitta

AU - Tinaburri, Lavinia

AU - D'Arcangelo, Daniela

AU - Sorice, Maurizio

AU - Capozzi, Antonella

AU - Garofalo, Tina

AU - Facchiano, Antonio

AU - Businaro, Rita

AU - Kumar, Prashant

AU - Singh, Brajendra K.

AU - Parmar, Virinder S.

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AU - Rigano, Rachele

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AB - We have previously demonstrated that human heat shock protein 90 (HSP90), an intracellular self protein, is the target of cellular and humoral autoimmune responses in patients with carotid atherosclerosis. In this study, we evaluated in vitro whether oxidative stress, a feature of atherosclerotic plaque, alters HSP90 expression in endothelial cells, thus inducing surface localization of this molecule and whether the antioxidant compound 7,8-dihydroxy-4-methylcoumarin (7,8-DHMC) is able to prevent oxidative stress-induced alterations of HSP90 localization. By the use of flow cytometry, immunofluorescence, enzyme-linked immunosorbent assay, and semiquantitative reverse-transcription polymerase chain reaction, we demonstrated that exposure of human umbilical vein endothelial cells (HUVEC) to the prooxidant compound H2O2 upregulated HSP90 surface expression and reduced its secretion without altering HSP90 gene expression and intracytoplasmic protein levels. Pretreatment of HUVEC with 7,8-DHMC prevented H2O2-induced alterations of HSP90 cellular distribution and secretion. Our results suggest that the strong oxidative conditions of atherosclerotic plaques promote the upregulation of HSP90 surface expression on endothelial cells, thus rendering the protein a possible target of autoimmune reactions. The antioxidant 7,8-DHMC, by preventing oxidative-stress-triggered HSP90 surface upregulation, may be useful to counteract possible autoreactive reactions to HSP90.

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