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Mol. Hum. Reprod. Advance Access published online on March 16, 2004

Molecular Human Reproduction, doi:10.1093/molehr/gah040
© 2004 by Oxford University Press
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Received November 21, 2003
Revised December 16, 2003
Accepted January 5, 2004

Article

Relaxin potentiates the expression of inducible nitric oxide synthase by endothelial cells from human umbilical vein in in vitro culture

S. Quattrone 1, L. Chiappini 1, G. Scapagnini 2, B. Bigazzi 1, D. Bani 3*

1 Department Anatomy, Histology & Forensic Medicine, Section of Histology, University of Florence, Florence, Italy
2 Department Experimental & Clinical Pharmacology, University of Catania, Catania, Italy
3 Department Anatomy, Histology & Forensic Medicine, Section of Histology, University of Florence, Florence, Italy; Dipartimento di Anatomia, Istologia e Medicina Legale. Sezione di Istologia, Università di Firenze, V.le G.Pieraccini, 6, I-50139 Florence, Italy

* To whom correspondence should be addressed. E-mail: daniele.bani{at}unifi.it.


   Abstract

The hormone relaxin (RLX), which can be detected in human venous cord blood, has been shown to be a potent vasodilator, acting through increased expression of inducible nitric oxide synthase (NOS II) and nitric oxide (NO) generation. This study aims at clarifying whether RLX, at concentrations of 100 and 1000 ng/ml for 6 or 12 h of exposure, can influence the expression of NOS isoforms in human umbilical vein endothelial cells (HUVEC) cultured in vitro. NOS mRNA expression was studied by quantitative real-time RT-PCR, NOS protein expression and activity was studied by Western blot and nitrite assay, and immunoreactive NOS localization was performed by confocal microscopy. Untreated HUVEC expressed all the NOS isoforms, especially the constitutive, endothelial-type NOS III and, to a lesser extent, NOS II and NOS I. RLX-treated cells showed an increased expression of NOS II, attaining a maximum with 1000 ng/ml RLX, which gave rise to increased NO generation, as shown by nitrite assay. This effect of RLX appears to be mediated by activation of NOS II transcription factor NF-kappaB, since it was abolished by the NF-kappaB inhibitors curcumin-95 and dexamethasone. These findings suggest that RLX in the umbilical vein might contribute to the NO-dependent regulation of vascular tone.

Key Words: Key words: human umbilical vein endothelial cells/relaxin/nitric oxide synthase I/NOS II/NOS III


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