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Cardiovasc Res. 2013 Aug 1;99(3):545-54. doi: 10.1093/cvr/cvt127. Epub 2013 May 25.

Shear stress is a positive regulator of thimet oligopeptidase (EC3.4.24.15) in vascular endothelial cells: consequences for MHC1 levels.

Author information

1
School of Biotechnology, Dublin City University, Glasnevin, Dublin 9, Ireland.

Abstract

AIMS:

Thimet oligopeptidase (TOP, endopeptidase EC3.4.24.15) is a soluble metallopeptidase known to be expressed within the mammalian vasculature. We examine for the first time the relationship between TOP expression and laminar shear stress, a haemodynamic force associated with endothelium-mediated vascular homeostasis.

METHODS AND RESULTS:

Human and bovine aortic endothelial cells were exposed to physiological levels of laminar shear (0-10 dynes/cm², 24-48 h) and monitored for TOP expression using promoter activity assay, qRT-PCR, western blotting, and immunocytochemistry. Using a luciferase reporter encoding the full-length rat TOP promoter, initial studies demonstrated shear-dependent promoter activation (~five-fold). TOP mRNA and protein were also consistently up-regulated by shear, events which could be completely prevented by pre-treatment of cells with either N-acetylcysteine, superoxide dismutase, or catalase, confirming ROS involvement. Consistent with this, targeted inhibition of NADPH oxidase (apocynin, NSC23766, NOX4 siRNA) had a similar blocking effect. Finally, in view of its pivotal role in cellular antigen presentation and major histocompatibility complex (MHC) class-1 regulation, we hypothesized that the shear-dependent induction of TOP may lower MHC1 expression. In this respect, we observed that recombinant TOP over-expression in static HAECs dose-dependently depleted MHC1 (>60%), while siRNA-mediated blockade of TOP induction in sheared HAECs led to substantially elevated MHC1 (~66%).

CONCLUSION:

Our findings demonstrate that laminar shear positively regulates endothelial TOP expression. Moreover, a role for ROS production by NADPH oxidase is indicated. Finally, our studies suggest that shear-dependent TOP induction down-regulates MHC1 levels, pointing to a role for TOP in the flow-mediated regulation of endothelial immunogenicity.

KEYWORDS:

Endothelium; MHC1; NADPH oxidase; Shear stress; Thimet oligopeptidase

PMID:
23708739
DOI:
10.1093/cvr/cvt127
[Indexed for MEDLINE]

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