Internal pudental artery dysfunction in diabetes mellitus is mediated by NOX1-derived ROS-, Nrf2-, and Rho kinase-dependent mechanisms

Alves-Lopes, Rhéure and Neves, Karla B. and Montezano, Augusto C. and Harvey, Adam and Carneiro, Fernando S. and Touyz, Rhian M. and Tostes, Rita C. (2016) Internal pudental artery dysfunction in diabetes mellitus is mediated by NOX1-derived ROS-, Nrf2-, and Rho kinase-dependent mechanisms. Hypertension, 68 (4). pp. 1056-1064. ISSN 1524-4563 (https://doi.org/10.1161/HYPERTENSIONAHA.116.07518)

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Abstract

Oxidative stress plays an important role in diabetes mellitus (DM)-associated vascular injury. DM is an important risk factor for erectile dysfunction. Functional and structural changes in internal pudendal arteries (IPA) can lead to erectile dysfunction. We hypothesized that downregulation of nuclear factor E2-related factor 2 (Nrf2), consequent to increased nicotinamide adenine dinucleotide phosphate oxidase 1 (NOX1)-derived reactive oxygen species (ROS), impairs IPA function in DM. IPA and vascular smooth muscle cells from C57BL/6 (control) and NOX1 knockout mice were used. DM was induced by streptozotocin in C57BL/6 mice. Functional properties of IPA were assessed using a myograph, protein expression and peroxiredoxin oxidation by Western blot, RNA expression by polymerase chain reaction, carbonylation by oxyblot assay, ROS generation by lucigenin, nitrotyrosine, and amplex red, and Rho kinase activity and nuclear accumulation of Nrf2 by ELISA. IPA from diabetic mice displayed increased contractions to phenylephrine (control 138.5±9.5 versus DM 191.8±15.5). ROS scavenger, Nrf2 activator, NOX1 and Rho kinase inhibitors normalized vascular function. High glucose increased ROS generation in IPA vascular smooth muscle cell. This effect was abrogated by Nrf2 activation and not observed in NOX1 knockout vascular smooth muscle cell. High glucose also increased levels of nitrotyrosine, protein oxidation/carbonylation, and Rho kinase activity, but reduced Nrf2 activity and expression of Nrf2-regulated genes (catalase [25.6±0.05%], heme oxygenase-1 [21±0.1%], and NAD(P)H:quinone oxidoreductase 1 [22±0.1%]) and hydrogen peroxide levels. These effects were not observed in vascular smooth muscle cell from NOX1 knockout mice. In these cells, high glucose increased hydrogen peroxide levels. In conclusion, Rho kinase activation, via NOX1-derived ROS and downregulation of Nrf2 system, impairs IPA function in DM. These data suggest that Nrf2 is vasoprotective in DM-associated erectile dysfunction.

ORCID iDs

Alves-Lopes, Rhéure, Neves, Karla B. ORCID logoORCID: https://orcid.org/0000-0001-5158-9263, Montezano, Augusto C., Harvey, Adam, Carneiro, Fernando S., Touyz, Rhian M. and Tostes, Rita C.;
  • Item type:Article
    ID code:82730
    Dates:
    Date
    Event
    1 October 2016
    Published
    15 August 2016
    Published Online
    17 July 2016
    Accepted
    Notes:Funding Information: Sources of Funding This study was funded by grants from FAPESP-Brazil (2012/12178- 6 to R. Alves-Lopes and 2013/08216-2 to the Center of Research on Inflammatory Diseases [CRID]), Science Without Borders-Brazil (BEX 2104/13-0), and the British Heart Foundation (CH/4/29762 and RG/13/7/30099). Publisher Copyright: © 2016 American Heart Association, Inc.
    Subjects:Medicine
    Department:Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences
    Depositing user: Pure Administrator
    Date deposited:13 Oct 2022 08:17
    Last modified:11 Nov 2024 13:39
    Related URLs:
    URI:https://strathprints.strath.ac.uk/id/eprint/82730
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