E Mitidieri1, V Vellecco1, V Brancaleone2, D Vanacore1, O L Manzo1, E Martin3, I Sharina3, Y Krutsenko4, M C Monti5, E Morretta5, A Papapetropoulos6,7, G Caliendo1, F Frecentese1, G Cirino1, R Sorrentino8, R d'Emmanuele di Villa Bianca1, M Bucci1. 1. Department of Pharmacy, School of Medicine and Surgery, University of Naples -Federico II-, Naples, Italy. 2. Department of Science, University of Basilicata, Potenza, Italy. 3. Department of Internal Medicine, Division of Cardiology, University of Texas Health Science Center, McGovern Medical School, Houston, Texas, USA. 4. School of Science and Technology, Nazarbayev University, Astana, Kazakhstan. 5. Department of Pharmacy, University of Salerno, Fisciano (SA), Italy. 6. Laboratory of Pharmacology, Department of Pharmacy, National and Kapodistrian University of Athens, Greece. 7. Clinical, Experimental Surgery and Translational Research Center, Biomedical Research Foundation of the Academy of Athens, Greece (A.P.). 8. Department of Molecular Medicine and Medical Biotechnologies, School of Medicine and Surgery, University of Naples -Federico II-, Naples, Italy.
Abstract
BACKGROUND AND PURPOSE: L-Cysteine or hydrogen sulfide (H2 S) donors induce a biphasic effect on pre-contracted isolated vessels. The contractile effect is displayed within a concentration range of 10nM-3μM followed by vasodilatation at 30-100μM. Here, we have investigated the signalling involved in H2 S-induced contraction. EXPERIMENTAL APPROACH: Vascular response to NaHS or L-cysteine is evaluated on isolated pre-contracted vessel rings harvested from wild type, cystathionine-γ-lyase (CSE-/- ), soluble guanylyl cyclase (sGCα1 -/- ) and endothelial nitric oxide synthase (eNOS-/- ) knock-out mice. The cAMP, cGMP and inosine 3',5'cyclic monophosphate (cIMP) levels are simultaneously quantified by using UPLC-MS/MS analysis. The involvement of sGC, phosphodiesterase (PDE) 4A and PDE5 are also evaluated. KEY RESULTS: CSE-derived H2 S-induced contraction requires an intact eNOS/NO/sGC pathway and involves cIMP as a second messenger. H2 S contractile effect involves a transient increase of cGMP and cAMP metabolism operated by PDE5 and PDE4A, thus unmasking cIMP contracting action. The stable cell-permeable analogue of cIMP elicits concentration dependent contraction on phenylephrine stable tone. The lack of cIMP, coupled to the hypo-contractility displayed by vessels harvested from CSE-/- mice, confirms that H2 S-induced contraction involves cIMP. CONCLUSION AND IMPLICATIONS: The endothelium dynamically regulates the vessel homeostasis also by modulating the contractile tone. This mechanism also involves CSE-derived H2 S that explicates its action through cIMP. This article is protected by copyright. All rights reserved.
BACKGROUND AND PURPOSE:L-Cysteine or hydrogen sulfide (H2 S) donors induce a biphasic effect on pre-contracted isolated vessels. The contractile effect is displayed within a concentration range of 10nM-3μM followed by vasodilatation at 30-100μM. Here, we have investigated the signalling involved in H2 S-induced contraction. EXPERIMENTAL APPROACH: Vascular response to NaHS or L-cysteine is evaluated on isolated pre-contracted vessel rings harvested from wild type, cystathionine-γ-lyase (CSE-/- ), soluble guanylyl cyclase (sGCα1 -/- ) and endothelial nitric oxide synthase (eNOS-/- ) knock-out mice. The cAMP, cGMP and inosine 3',5'cyclic monophosphate (cIMP) levels are simultaneously quantified by using UPLC-MS/MS analysis. The involvement of sGC, phosphodiesterase (PDE) 4A and PDE5 are also evaluated. KEY RESULTS:CSE-derived H2 S-induced contraction requires an intact eNOS/NO/sGC pathway and involves cIMP as a second messenger. H2 S contractile effect involves a transient increase of cGMP and cAMP metabolism operated by PDE5 and PDE4A, thus unmasking cIMP contracting action. The stable cell-permeable analogue of cIMP elicits concentration dependent contraction on phenylephrine stable tone. The lack of cIMP, coupled to the hypo-contractility displayed by vessels harvested from CSE-/- mice, confirms that H2 S-induced contraction involves cIMP. CONCLUSION AND IMPLICATIONS: The endothelium dynamically regulates the vessel homeostasis also by modulating the contractile tone. This mechanism also involves CSE-derived H2 S that explicates its action through cIMP. This article is protected by copyright. All rights reserved.