Analyses of the anti-inflammatory protein galectin-1 in the human retinal pigment epithelial cells after activation by endotoxin

• Grant number: 11/05248-5
• Support Opportunities: Scholarships in Brazil - Master
• Start date: March 01, 2012
• End date: February 28, 2013
• Field of knowledge: Biological Sciences - Morphology - Histology
• Principal Investigator: Sonia Maria Oliani
• Grantee: Nathália Martins Sonehara
• Host Institution: Instituto de Biociências, Letras e Ciências Exatas (IBILCE). Universidade Estadual Paulista (UNESP). Campus de São José do Rio Preto. São José do Rio Preto , SP, Brazil

Abstract

Inflammation is the main contributing factor for many eye disorders and may lead to decreased vision and even blindness. In the treatment of intraocular inflammation in general, glucocorticoids are frequently administered medications. However, the collateral effect of these pharmaco, as the rise of the pressure to intraocular, cataractogenesis and potential cytotoxicity to the structures of the retina, limit its uses. Because of these restrictions, there is an obvious demand for the development of new therapeutic strategies. Among the anti-inflammatory mediators, including protein endogenous galectin-1 (Gal-1) capable of controlling the process of leukocyte transmigration and apoptosis, contributing to the homeostasis of the inflammatory reaction. However, the expression of Gal-1 in normal and inflamed ocular tissues has been poorly studied. For these reasons, this study aims to evaluate in vitro the anti-inflammatory effect of the administration of Gal-1 protein. We investigate the expression and localization of endogenous Gal-1 in ARPE-19 cells (derived from human retinal pigment epithelial normal) after activation by bacterial lipopolysaccharide (LPS), and the possible anti-inflammatory effect of the administration of protein in these cells. Initially, the ARPE-19 cells are grown in DMEM: F-12, the concentration of 104células/35mm per sample, and divided into the following groups: control, activated by LPS and activated by LPS / Gal-1 treated with recombinant human ( hrGal-1, 0.04 to 4 mg / mL) or dexametaxona (Dex; 1mm). To determine the specificity of the effect of hrGal-1 will be added in some wells Beta-galactose sugar or sucrose (100 mM). Investigations are held in the 0, 2, 8, 24 and 48 hours. Groups in LPS, and LPS LPS/Gal-1 / Dex cells will be processed for analysis of the expression of COX-2 by immunofluorescence. The endogenous Gal-1 protein is evaluated by Western blotting in LPS group and LPS / Dex. The supernatants obtained in all experimental conditions, will be collected for measurements of pro-inflammatory cytokines TNF-alfa, IL-6, IL-8, IL-10 and MCP1 by MAGPIX System. Investigations will be conducted in view of the applications of anti-inflammatory protein Gal-1 as a therapeutic alternative eye.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ZANON, CAROLINE DE FREITAS; SONEHARA, NATHALIA MARTINS; GIROL, ANA PAULA; GIL, CRISTIANE DAMAS; OLIANI, SONIA MARIA. Protective effects of the galectin-1 protein on in vivo and in vitro models of ocular inflammation. MOLECULAR VISION, v. 21, p. 1036-1050, SEP 2 2015. (11/21845-3, 11/05248-5, 12/02759-1)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

SONEHARA, Nathália Martins. Avaliação do efeito inflamatório da proteína galectina-1 nas células epiteliais pigmentadas da retina humana (ARPE-19) após ativação por endotoxina. 2013. Master's Dissertation - Universidade Estadual Paulista (Unesp). Instituto de Biociências Letras e Ciências Exatas. São José do Rio Preto São José do Rio Preto.