title: ANXA1Ac2-26peptide, a possible therapeutic approach in inflammatory ocular diseases.
abstract: The eye is immunologically privileged when inflammatory responses are suppressed. One component responsible for the suppression of inflammatory responses is the blood retinal barrier, which comprises the retinal pigment epithelium. The destruction of this barrier initiates inflammation, which can affect any part of the eye. Therefore, inflammatory response is controlled by the action of anti-inflammatory mediators, among these mediators, annexin A1 (ANXA1) protein acts as a modulator of inflammation. In this study we aimed to improve the knowledge of this area by investigating how a peptide of the ANXA1 protein (ANXA1Ac2-26) modulates the morphology, proliferation, migration and expression of genes and proteins in human retinal pigment epithelium cells (ARPE-19). Determining how signaling pathways (NF-κB and UBC) are modulated by the ANXA1Ac2-26peptide could be important for understanding the inflammatory process. ARPE-19 cells were activated by bacterial lipopolysaccharide endotoxin (LPS) and treated with ANXA1Ac2-26peptide, in a concentration of 1.7μM and 33.8μM. We observed that the LPS activation diminished the levels of endogenous ANXA1 after 2h and 24h and ANXA1Ac2-26peptide decreased the proliferation and re-establishes the migration of ARPE-19 cells. After using a hybridization approach, 80 differentially expressed genes were found. Five of these genes were selected (LRAT, CTGF, MAP1B, ALDH1A3 and SETD7) and all were down-regulated after treatment with the peptide. The genes CTGF and LRAT would be considered as potential molecular markers of ophthalmologic inflammation. The expression of pro-inflammatory cytokines was also decreased after the treatment, indicating the efficiency of the anti-inflammatory peptide at high concentrations, since the reduction in the levels of these mediators were observed after the treatment with ANXA1Ac2-26peptide at 33.8μM. Our results suggest that the retinal pigment epithelial cells are a potential target of the ANXA1 protein and point to possible applications of the ANXA1Ac2-26peptide as an innovative therapy for the treatment of ocular inflammation.
authors.affiliation: Department of Biology, Institute of Biosciences, Letters and Science - IBILCE/UNESP, São José do Rio Preto, SP, Brazil.
authors.forename: Laila Toniol
authors.initials: LT
authors.lastname: Cardin
authors.affiliation: Department of Biology, Institute of Biosciences, Letters and Science - IBILCE/UNESP, São José do Rio Preto, SP, Brazil.
authors.forename: Nathália Martins
authors.initials: NM
authors.lastname: Sonehara
authors.affiliation: Department of Biology, Institute of Biosciences, Letters and Science - IBILCE/UNESP, São José do Rio Preto, SP, Brazil.
authors.forename: Kallyne Kioko Oliveira
authors.initials: KK
authors.lastname: Mimura
authors.affiliation: Department of Clinical Medical, Foundation Blood Center of Ribeirão Preto, Faculty of Medicine of Ribeirão Preto, University of São Paulo - FCFRP/USP, Ribeirão Preto, SP, Brazil.
authors.forename: Anemari
authors.initials: A
authors.lastname: Ramos Dinarte Dos Santos
authors.affiliation: Department of Clinical Medical, Foundation Blood Center of Ribeirão Preto, Faculty of Medicine of Ribeirão Preto, University of São Paulo - FCFRP/USP, Ribeirão Preto, SP, Brazil.
authors.forename: Wilson Araújo
authors.initials: WA
authors.lastname: da Silva
authors.affiliation: Department of Clinical Analyses, Toxicology and Food Science, Faculty of Pharmaceutical Science of Ribeirão Preto, University of São Paulo - FCFRP/USP, Ribeirão Preto, SP, Brazil.
authors.forename: Lays Martin
authors.initials: LM
authors.lastname: Sobral
authors.affiliation: Department of Clinical Analyses, Toxicology and Food Science, Faculty of Pharmaceutical Science of Ribeirão Preto, University of São Paulo - FCFRP/USP, Ribeirão Preto, SP, Brazil.
authors.forename: Andréia Machado
authors.initials: AM
authors.lastname: Leopoldino
authors.affiliation: Department of Molecular Biology, Faculty of Medicine of São José do Rio Preto - FAMERP, São José do Rio Preto, SP, Brazil.
authors.forename: Bianca Rodrigues
authors.initials: BR
authors.lastname: da Cunha
authors.affiliation: Department of Molecular Biology, Faculty of Medicine of São José do Rio Preto - FAMERP, São José do Rio Preto, SP, Brazil.
authors.forename: Eloiza H
authors.initials: EH
authors.lastname: Tajara
authors.affiliation: Department of Biology, Institute of Biosciences, Letters and Science - IBILCE/UNESP, São José do Rio Preto, SP, Brazil.
authors.forename: Sonia Maria
authors.initials: SM
authors.lastname: Oliani
authors.affiliation: Department of Biology and Animal Science, Faculty of Engineering of Ilha Solteira - FEIS/UNESP, Ilha Solteira, SP, Brazil. Electronic address: lisoni@bio.feis.unesp.br.
authors.forename: Flávia Cristina
authors.initials: FC
authors.lastname: Rodrigues-Lisoni
mesh_terms: D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism; D017305:Annexin A1 / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D015153:Blotting, Western; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects* / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects* / Q000235:genetics; D055513:Connective Tissue Growth Factor / Q000235:genetics / Q000378:metabolism; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D005128:Eye Diseases / Q000235:genetics / Q000517:prevention & control; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling / Q000379:methods; D053263:Gene Regulatory Networks / Q000187:drug effects; D006801:Humans; D007249:Inflammation / Q000235:genetics / Q000517:prevention & control; D008070:Lipopolysaccharides / Q000494:pharmacology; D010455:Peptides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D055213:Retinal Pigment Epithelium / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D017305:Annexin A1; D000893:Anti-Inflammatory Agents; C526317:CCN2 protein, human; D016207:Cytokines; D008070:Lipopolysaccharides; D010455:Peptides; C096153:annexin A1 peptide (2-26); D055513:Connective Tissue Growth Factor; D000217:Acyltransferases; C056942:lecithin-retinol acyltransferase