title: Arginase-2 is cooperatively up-regulated by nitric oxide and histone deacetylase inhibition in human umbilical artery endothelial cells.
abstract: Arginase-2 counteracts endothelial nitric oxide synthase (eNOS) activity in human endothelium, and its expression is negatively controlled by histone deacetylase (HDAC2). Conversely NO inhibits HDAC and previous studies suggest that arginase-2 is up-regulated by NO. We studied whether NO regulates arginase-2 expression in umbilical artery endothelial cells (HUAEC) increasing ARG2 promoter accessibility. HUAEC exposed to NOC-18 (NO donor, 1-100 μM, 0-24 h) showed an increase in arginase-2 but a decrease in eNOS mRNA levels in a time-dependent manner, with a maximal effect at 100 μM (24 h). Conversely NOS inhibition with L-NAME (100 μM) reduced arginase-2 mRNA and protein levels, an effect reverted by co-incubation with NOC-18. Treatment with TSA paralleled the effects of NO on arginase-2 and eNOS at mRNA and protein levels, with maximal effect at 10 μM. Co-incubation of NOC-18 (100 μM) with a sub-maximal concentration of TSA (1 μM) potentiated the increase in arginase-2 mRNA levels, whilst L-NAME prevented TSA-dependent arginase-2 induction. The effects on arginase-2 mRNA were paralleled by changes in chromatin accessibility, as well as increased levels of H3K9 and H4K12 acetylation, at ARG2 proximal (-579 to -367 and -280 to -73 bp from TSS) and core (-121 to +126 bp from TSS) promoter. Finally NO-dependent arginase-2 induction was prevented by pre-incubation for 10 min with the cysteine blocker MMTS (10 mM). These data showed for the first time that NO up-regulates arginase-2 expression in primary cultured human endothelial cells by an epigenetic-mediated mechanism increasing ARG2 promoter accessibility suggesting a negative regulatory loop for eNOS activity.
authors.affiliation: Division of Obstetrics & Gynecology, Faculty of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile. Electronic address: bjkrause@uc.cl.
authors.forename: Bernardo J
authors.initials: BJ
authors.lastname: Krause
authors.affiliation: Division of Obstetrics & Gynecology, Faculty of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile.
authors.forename: Cherie
authors.initials: C
authors.lastname: Hernandez
authors.affiliation: Division of Obstetrics & Gynecology, Faculty of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile.
authors.forename: Andres
authors.initials: A
authors.lastname: Caniuguir
authors.affiliation: Division of Obstetrics & Gynecology, Faculty of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile.
authors.forename: Paola
authors.initials: P
authors.lastname: Vasquez-Devaud
authors.affiliation: Division of Obstetrics & Gynecology, Faculty of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile.
authors.forename: Ivo
authors.initials: I
authors.lastname: Carrasco-Wong
authors.affiliation: Division of Pediatrics, Faculty of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Uauy
authors.affiliation: Division of Obstetrics & Gynecology, Faculty of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile; Division of Pediatrics, Faculty of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile. Electronic address: pcasane@uc.cl.
authors.forename: Paola
authors.initials: P
authors.lastname: Casanello
mesh_terms: D001119:Arginase / Q000096:biosynthesis*; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D056572:Histone Deacetylase Inhibitors / Q000378:metabolism* / Q000494:pharmacology; D006655:Histone Deacetylases / Q000378:metabolism; D006801:Humans; D019331:NG-Nitroarginine Methyl Ester / Q000378:metabolism / Q000494:pharmacology; D009569:Nitric Oxide / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D014469:Umbilical Arteries / Q000187:drug effects / Q000378:metabolism*; D015854:Up-Regulation / Q000502:physiology
chemical_list: D056572:Histone Deacetylase Inhibitors; D009569:Nitric Oxide; D006655:Histone Deacetylases; C477631:ARG2 protein, human; D001119:Arginase; D019331:NG-Nitroarginine Methyl Ester
keywords: Chromatin accessibility; DETA NONOate (PubChem CID: 1606); HDAC; Histone acetylation; Human endothelial cells; N(G)-Nitro-l-arginine methyl ester (PubChem CID: 39836); Nitric oxide arginase-2; S-Methyl methanethiosulfonate (PubChem CID: 18064); Trichostatin A (PubChem CID: 444732)