The Metabolism of Epoxyeicosatrienoic Acids by Soluble Epoxide Hydrolase Is Protective against the Development of Vascular Calcification.

Olivier Varennes,Thibaut Objois,Thomas Duflot,Christophe Morisseau,Carine Avondo,Saïd Kamel,Isabelle Six,Gaëlle Lenglet,Romuald Mentaverri,Michel Brazier,Jeremy Bellien,Gilles Kauffenstein

doi:10.3390/ijms21124313

Abstract

This study addressed the hypothesis that soluble epoxide hydrolase (sEH), which metabolizes endothelium-derived epoxyeicosatrienoic acids, plays a role in vascular calcification. The sEH inhibitor trans-4-(4-(3-adamantan-1-yl-ureido)-cyclohexyloxy)-benzoic acid (t-AUCB) potentiated the increase in calcium deposition of rat aortic rings cultured in high-phosphate conditions. This was associated with increased tissue-nonspecific alkaline phosphatase activity and mRNA expression level of the osteochondrogenic marker Runx2. The procalcifying effect of t-AUCB was prevented by mechanical aortic deendothelialization or inhibition of the production and action of epoxyeicosatrienoic acids using the cytochrome P450 inhibitor fluconazole and the antagonist 14,15-epoxyeicosa-5(Z)-enoic acid (14,15-EEZE), respectively. Similarly, exogenous epoxyeicosatrienoic acids potentiated the calcification of rat aortic rings through a protein kinase A (PKA)-dependent mechanism and of human aortic vascular smooth muscle cells when sEH was inhibited by t-AUCB. Finally, a global gene expression profiling analysis revealed that the mRNA expression level of sEH was decreased in human carotid calcified plaques compared to adjacent lesion-free sites and was inversely correlated with Runx2 expression. These results show that sEH hydrolase plays a protective role against vascular calcification by reducing the bioavailability of epoxyeicosatrienoic acids.

Highlights

Vascular calcification is a highly regulated process observed during aging, hypertension, diabetes, and chronic kidney disease (CKD)
There was an increase in tissue-nonspecific alkaline phosphatase (TNAP) activity (Figure 1F) and a decrease in pyrophosphate (PPi) level (Figure 1G), an endogenous inhibitor of ectopic calcification which is hydrolyzed by TNAP [16,17,18], in the culture supernatants of high-phosphate-cultured aortic rings
T3h.8emdMecrPeai saenidnaSMcuMltuHrCe dmuRraNtiAonexopf r7esdsaioyns ilsevneeleadnedd tthoeinindcurceeasseiginniRficuannxt2a, oMrtsixc2caalncdifiScoatxi9onle[v1e6l]s. cTohnefidrmecerdeatsheeintrSaMnsMitiHonC omf RvNasAcuelxapr rceeslslsiotnowlevaredl aanndotshteeoincchroenadseroigneRnuicnxp2h,eMnostxy2paen. dInSoaxd9dlietvioenls, tchoenrfeirwmaesd athneitnrcarnesaitsieodn TofNvAasPcualcatrivcietlylsatnodwadredcraenaosesteinocthhoenldervoeglsenoifc pthheeninohtyibpiet.oIrnoafdcdailtcioifinc, athtieorne PwPais, aasn pinrecrveiaosuesdlyTsNhAowPnacitnivaitoyrtaansdisdoelactreedasferoinmthuerelmevicelsraotsf t[h1e4].inhTihbiistopraotfhwcaalcyifiiscaitniovnolvPePdi, ians mineralization deposition facilitating the formation of the final product of calcium phosphate salt reaction hydroxyapatite, but it may directly promote the transition of vascular smooth muscle cells (VSMCs) to a osteochondrogenic phenotype [31,32]

Summary

Introduction

Vascular calcification is a highly regulated process observed during aging, hypertension, diabetes, and chronic kidney disease (CKD). It contributes to elevated risk of cardiovascular events and mortality [1,2,3,4]. Increasing experimental evidence shows that the endothelium is a source of osteoprogenitor cells and regulates vascular calcification through the release of signaling molecules such as nitric oxide (NO), which prevents VSMC osteochondrogenic differentiation [5,6,7]. Based on the measurements of plasma DHET levels, a recent clinical study suggested that EETs may be protective against the development of vascular calcification [13]

Objectives

Methods

Results

Conclusion