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Hydrogen sulfide attenuates calcification of vascular smooth muscle cells via KEAP1/NRF2/NQO1 activation

Hydrogen sulfide attenuates calcification of vascular smooth muscle cells via KEAP1/NRF2/NQO1 activation
Hydrogen sulfide attenuates calcification of vascular smooth muscle cells via KEAP1/NRF2/NQO1 activation
Background and aims:

Vascular calcification is a common health problem related to oxidative stress, inflammation, and circulating calciprotein particles (CPP). Hydrogen sulfide is an endogenous signaling molecule with antioxidant properties and potential for drug development targeting redox signaling. Yet, its molecular mechanisms of action in vascular smooth muscle cell (VSMC) calcification have not been delineated. We therefore sought to identify key pathways involved in the calcification-inhibitory properties of sulfide employing our recently developed CPP-induced VSMC calcification model.

Methods:

Using next-generation sequencing, we investigated the transcriptomic changes of sodium hydrosulfide-treated versus non-treated calcifying VSMCs. The potential role of candidate genes and/or regulatory pathways in prevention of calcification was investigated by small interfering RNA (siRNA).

Results:

CPP led to a pronounced accumulation of cell-associated calcium, which was decreased by sulfide in a concentration-dependent manner. Both, CPP-induced hydrogen peroxide production and enhanced pro-inflammatory/oxidative stress-related gene expression signatures were attenuated by sulfide-treatment. Gene ontology enrichment and in silico pathway analysis of our transcriptome data suggested NAD(P)H dehydrogenase [quinone] 1 (NQO1) as potential mediator. Corroborating these findings, silencing of Kelch-like ECH-associated protein 1 (KEAP1), an inhibitor of nuclear factor (erythroid-derived 2)-like 2 (NRF2) nuclear activity, enhanced NQO1 expression, whereas NRF2 silencing reduced the expression of NQO1 and abrogated the calcification-suppressing activity of sulfide. Moreover, immunofluorescence microscopy and Western blot analysis confirmed nuclear translocation of NRF2 by sulfide in VSMC.

Conclusions:

Sulfide attenuates CPP-induced VSMC calcification in vitro via the KEAP1-NRF2 redox sensing/stress response system by enhancing NQO1 expression.
0021-9150
78-86
Aghagolzadeh, Parisa
d3d7277c-6f8b-49be-9ce6-9eafdcd204d5
Radpour, Ramin
4be8463d-84d9-49ea-972c-b1cbe93c24c7
Bachtler, Matthias
e84b3d87-6978-48da-be8e-49bcdf1f8ca1
van Goor, Harry
6e4f96a5-c749-43b6-a488-6af71f932dc3
Smith, Edward R.
00caceb0-ebdc-4274-b05e-fc1f6a1c8113
Lister, Adam
bc5c3344-bdcf-45cd-b22b-7c9b9151c744
Odermatt, Alex
4ea0e54a-2c68-4952-9c66-025a6438bd9c
Feelisch, Martin
8c1b9965-8614-4e85-b2c6-458a2e17eafd
Pasch, Andreas
5f5d9b34-4d9e-40a4-9326-fa1cd352717d
Aghagolzadeh, Parisa
d3d7277c-6f8b-49be-9ce6-9eafdcd204d5
Radpour, Ramin
4be8463d-84d9-49ea-972c-b1cbe93c24c7
Bachtler, Matthias
e84b3d87-6978-48da-be8e-49bcdf1f8ca1
van Goor, Harry
6e4f96a5-c749-43b6-a488-6af71f932dc3
Smith, Edward R.
00caceb0-ebdc-4274-b05e-fc1f6a1c8113
Lister, Adam
bc5c3344-bdcf-45cd-b22b-7c9b9151c744
Odermatt, Alex
4ea0e54a-2c68-4952-9c66-025a6438bd9c
Feelisch, Martin
8c1b9965-8614-4e85-b2c6-458a2e17eafd
Pasch, Andreas
5f5d9b34-4d9e-40a4-9326-fa1cd352717d

Aghagolzadeh, Parisa, Radpour, Ramin, Bachtler, Matthias, van Goor, Harry, Smith, Edward R., Lister, Adam, Odermatt, Alex, Feelisch, Martin and Pasch, Andreas (2017) Hydrogen sulfide attenuates calcification of vascular smooth muscle cells via KEAP1/NRF2/NQO1 activation Atherosclerosis, 265, pp. 78-86. (doi:10.1016/j.atherosclerosis.2017.08.012).

Record type: Article

Abstract

Background and aims:

Vascular calcification is a common health problem related to oxidative stress, inflammation, and circulating calciprotein particles (CPP). Hydrogen sulfide is an endogenous signaling molecule with antioxidant properties and potential for drug development targeting redox signaling. Yet, its molecular mechanisms of action in vascular smooth muscle cell (VSMC) calcification have not been delineated. We therefore sought to identify key pathways involved in the calcification-inhibitory properties of sulfide employing our recently developed CPP-induced VSMC calcification model.

Methods:

Using next-generation sequencing, we investigated the transcriptomic changes of sodium hydrosulfide-treated versus non-treated calcifying VSMCs. The potential role of candidate genes and/or regulatory pathways in prevention of calcification was investigated by small interfering RNA (siRNA).

Results:

CPP led to a pronounced accumulation of cell-associated calcium, which was decreased by sulfide in a concentration-dependent manner. Both, CPP-induced hydrogen peroxide production and enhanced pro-inflammatory/oxidative stress-related gene expression signatures were attenuated by sulfide-treatment. Gene ontology enrichment and in silico pathway analysis of our transcriptome data suggested NAD(P)H dehydrogenase [quinone] 1 (NQO1) as potential mediator. Corroborating these findings, silencing of Kelch-like ECH-associated protein 1 (KEAP1), an inhibitor of nuclear factor (erythroid-derived 2)-like 2 (NRF2) nuclear activity, enhanced NQO1 expression, whereas NRF2 silencing reduced the expression of NQO1 and abrogated the calcification-suppressing activity of sulfide. Moreover, immunofluorescence microscopy and Western blot analysis confirmed nuclear translocation of NRF2 by sulfide in VSMC.

Conclusions:

Sulfide attenuates CPP-induced VSMC calcification in vitro via the KEAP1-NRF2 redox sensing/stress response system by enhancing NQO1 expression.

Text Aghagolzade et al., (revised manuscript R2-2)_AP Kopie - Accepted Manuscript
Restricted to Repository staff only until 19 August 2018.

More information

Accepted/In Press date: 17 August 2017
e-pub ahead of print date: 19 August 2017
Published date: October 2017

Identifiers

Local EPrints ID: 414787
URI: http://eprints.soton.ac.uk/id/eprint/414787
ISSN: 0021-9150
PURE UUID: dae405ee-6014-4f94-be8f-337ebb0ce51b
ORCID for Martin Feelisch: ORCID iD orcid.org/0000-0003-2320-1158

Catalogue record

Date deposited: 11 Oct 2017 16:31
Last modified: 23 Oct 2017 16:30

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Contributors

Author: Parisa Aghagolzadeh
Author: Ramin Radpour
Author: Matthias Bachtler
Author: Harry van Goor
Author: Edward R. Smith
Author: Adam Lister
Author: Alex Odermatt
Author: Martin Feelisch ORCID iD
Author: Andreas Pasch

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