title: Identification of intracellular proteins and signaling pathways in human endothelial cells regulated by angiotensin-(1-7).
abstract: The study aimed to identify proteins regulated by the cardiovascular protective peptide angiotensin-(1-7) and to determine potential intracellular signaling cascades. Human endothelial cells were stimulated with Ang-(1-7) for 1 h, 3 h, 6 h, and 9 h. Peptide effects on intracellular signaling were assessed via antibody microarray, containing antibodies against 725 proteins. Bioinformatics software was used to identify affected intracellular signaling pathways. Microarray data was verified exemplarily by Western blot, Real-Time RT-PCR, and immunohistochemical studies. The microarray identified 110 regulated proteins after 1 h, 119 after 3 h, 31 after 6 h, and 86 after 9 h Ang-(1-7) stimulation. Regulated proteins were associated with high significance to several metabolic pathways like “Molecular Mechanism of Cancer” and “p53 signaling” in a time dependent manner. Exemplarily, Western blots for the E3-type small ubiquitin-like modifier ligase PIAS2 confirmed the microarray data and displayed a decrease by more than 50% after Ang-(1-7) stimulation at 1 h and 3 h without affecting its mRNA. Immunohistochemical studies with PIAS2 in human endothelial cells showed a decrease in cytoplasmic PIAS2 after Ang-(1-7) treatment. The Ang-(1-7) mediated decrease of PIAS2 was reproduced in other endothelial cell types. The results suggest that angiotensin-(1-7) plays a role in metabolic pathways related to cell death and cell survival in human endothelial cells.
authors.affiliation: Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, Universität Heidelberg, Germany; Department of Pharmacology and Therapeutics, University College Cork, Cork, Ireland.
authors.forename: Christian
authors.initials: C
authors.lastname: Meinert
authors.affiliation: Division of Nephrology, Department of Internal Medicine III, Faculty of Medicine, Technische Universität Dresden, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Gembardt
authors.affiliation: Department of Obstetrics, Division of Women and Child Health, Universität Leipzig, Leipzig, Germany.
authors.forename: Ilka
authors.initials: I
authors.lastname: Böhme
authors.affiliation: Department of Pharmacology and Therapeutics, University College Cork, Cork, Ireland; Department of Obstetrics, Division of Women and Child Health, Universität Leipzig, Leipzig, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Tetzner
authors.affiliation: Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, Universität Heidelberg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Wieland
authors.affiliation: Division of Cardiology, University of California, San Diego, USA.
authors.forename: Barry
authors.initials: B
authors.lastname: Greenberg
authors.affiliation: Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, Universität Heidelberg, Germany; Department of Pharmacology and Therapeutics, University College Cork, Cork, Ireland; Department of Obstetrics, Division of Women and Child Health, Universität Leipzig, Leipzig, Germany. Electronic address: thomas.walther2011@gmail.com.
authors.forename: Thomas
authors.initials: T
authors.lastname: Walther
mesh_terms: D000803:Angiotensin I / Q000378:metabolism*; D016923:Cell Death; D002470:Cell Survival; D015536:Down-Regulation; D042783:Endothelial Cells / Q000378:metabolism; D005786:Gene Expression Regulation; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D007150:Immunohistochemistry; D010446:Peptide Fragments / Q000378:metabolism*; D040081:Protein Array Analysis; D011506:Proteins / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism; D040901:Proteomics; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D010446:Peptide Fragments; D011506:Proteins; D020543:Proteome; D000803:Angiotensin I; C118790:angiotensin I (1-7)
keywords: Angiotensin-(1-7); Antibody microarray; Cell signaling; Endothelium; Renin-angiotensin system