title: TNFα induces the expression of genes associated with endothelial dysfunction through p38MAPK-mediated down-regulation of miR-149.
abstract: MicroRNAs have been proposed as novel regulators of vascular inflammation and dysfunction. This study aimed to evaluate the role of miR-149 in regulating the expression of key molecules associated with TNFα-induced endothelial activation. miR-149 was selected by in silico analysis and microRNA target prediction. Endothelial dysfunction was induced by TNFα treatment in Eahy926 endothelial cells and HUVEC. miR-149 level was evaluated by quantitative real time-polymerase chain reaction (RT-qPCR). Metalloproteinase-9 (MMP-9) was measured by zymography, Inducible Nitric Oxide Synthase (iNOS) by immunoblotting, Interleukin-6 (IL-6) and Interleukin-8 (IL-8) by ELISA. miR-149 regulatory effect was evaluated by gain-of-function technique upon miR-149 mimics transfection. TNFα down-modulated miR-149 level in Eahy926 and HUVEC. This effect was significantly abolished in Eahy926 by treatment with p38MAPK inhibitor. miR-149 mimic transfection counteracted the TNFα-induced expression of MMP-9, iNOS and IL-6. No effect was detected on IL-8 expression. Our results suggest that miR-149 represents an important new regulator of endothelial function through negative regulation of molecules associated with TNFα-induced endothelial dysfunction.
authors.affiliation: Vascular and Endovascular Surgery Unit, Research Laboratory of Experimental and Clinical Vascular Biology, DISC, University of Genoa, IRCCS San Martino - IST, 16132 Genoa, Italy. Electronic address: danielapalmieri@yahoo.com.
authors.forename: Daniela
authors.initials: D
authors.lastname: Palmieri
authors.affiliation: Department of Neuroscience, Rehabilitation, Ophthalmology, Genetics and Maternal and Child Health-Section of Medical Genetics, University of Genoa, IRCCS San Martino - IST, 16132 Genoa, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Capponi
authors.affiliation: Department of Neuroscience, Rehabilitation, Ophthalmology, Genetics and Maternal and Child Health-Section of Medical Genetics, University of Genoa, IRCCS San Martino - IST, 16132 Genoa, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Geroldi
authors.affiliation: Vascular and Endovascular Surgery Unit, Research Laboratory of Experimental and Clinical Vascular Biology, DISC, University of Genoa, IRCCS San Martino - IST, 16132 Genoa, Italy.
authors.forename: Marzia
authors.initials: M
authors.lastname: Mura
authors.affiliation: Department of Neuroscience, Rehabilitation, Ophthalmology, Genetics and Maternal and Child Health-Section of Medical Genetics, University of Genoa, IRCCS San Martino - IST, 16132 Genoa, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Mandich
authors.affiliation: Vascular and Endovascular Surgery Unit, Research Laboratory of Experimental and Clinical Vascular Biology, DISC, University of Genoa, IRCCS San Martino - IST, 16132 Genoa, Italy.
authors.forename: Domenico
authors.initials: D
authors.lastname: Palombo
mesh_terms: D002478:Cells, Cultured; D015536:Down-Regulation; D005786:Gene Expression Regulation; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D015850:Interleukin-6 / Q000235:genetics*; D020780:Matrix Metalloproteinase 9 / Q000235:genetics*; D035683:MicroRNAs / Q000235:genetics / Q000502:physiology*; D052247:Nitric Oxide Synthase Type II / Q000235:genetics*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology / Q000502:physiology*; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: C508600:IL6 protein, human; D015850:Interleukin-6; C551557:MIRN149 microRNA, human; D035683:MicroRNAs; D014409:Tumor Necrosis Factor-alpha; C496319:NOS2 protein, human; D052247:Nitric Oxide Synthase Type II; D048051:p38 Mitogen-Activated Protein Kinases; D020780:Matrix Metalloproteinase 9
keywords: Endothelial dysfunction; IL-6; MMP-9; MicroRNA; TNFα; iNOS; miR-149