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Endotoxin-induced uveitis: analysis of the interaction Annexin A1 and natural bioactive extracts as an alternative therapy

Grant number: 15/03359-5
Support type:Regular Research Grants
Duration: September 01, 2015 - August 31, 2017
Field of knowledge:Biological Sciences - Morphology - Cytology and Cell Biology
Principal researcher:Ana Paula Girol
Grantee:Ana Paula Girol
Home Institution: Faculdades Integradas Padre Albino (FIPA). Fundação Padre Albino (FPA). Catanduva , SP, Brazil
Assoc. researchers:Sonia Maria Oliani

Abstract

Uveitis is associated with various infectious and autoimmune diseases and is one of the leading causes of blindness worldwide. The different drugs used in the treatment of uveitis have severe side effects, which stimulates the search for new therapeutic alternatives. In recent years, the role of anti-inflammatory protein annexin A1 (AnxA1) in ocular inflammatory processes has been investigated by our group, using in vivo and in vitro studies, and point out the AnxA1 as a key mediator in the homeostasis of the inflammatory process. Other studies also indicate the anti-inflammatory action of natural bioactive extracts (NBEs) and recently it was demonstrated the interaction of AnxA1 and piplartine, a pepper NBE. In view of these considerations and because there are no studies on the action of NBEs in uveitis, the aim of this study is to investigate the possible anti-inflammatory effect of the administration of NBE separately or in combination with the mimetic peptide Ac2-26 of AnxA1, in the endotoxin induced uveitis (EIU). Male wistar rats (150-200 g) will be divided into five groups (n=10/group). For the development of EIU, four groups of animals will be anesthetized and inoculated into the right paw with 1 mg/kg of lipopolysaccharide (LPS) in 100 µl PBS. Animals without manipulation will be used as controls. Three EIU groups will be treated intraperitoneally, 15 minutes after LPS induction, to evaluate the anti-inflammatory effects of the Ac2-26 (1 mg/kg in 100 µl PBS), pepper NBE (1 mg/kg in 100 µl PBS) and Ac2-26/NBE (1 mg /kg of Ac2-26 + 1 mg/kg of NBE in 100 µL of PBS). The EIU animals, treated or not, will be sacrificed after 24 hours of LPS inoculation, by overdose of anesthetic, to collect blood, aqueous humor (AqH) and the eyes. The efficacy of the NBE and Ac2-26, associated or not, will be evaluated by histopathology and quantitative analyzes of neutrophils in the AqH, and macrophages in ocular tissues after immunohistochemistry. The immunohistochemical method will also be used to verify the expression of AnxA1, formyl peptide receptor (fpr)2 and cyclooxygenase 2 in ocular tissues. The gene expression of Anxa1 and fpr2 will be analyzed by RT-PCR. The supernatants from ocular tissues after maceration and blood plasma will be used for the dosage of the cytokines, tumor necrosis factor-±, monocyte chemotactic protein 1, interleukin (IL)1², , IL6 and IL10 using the Milliplex MAP panels. This research will allow the understanding of Ac2-26/NBE interaction and will also strengthen the AnxA1 studies in ocular inflammation, which may lead to new therapeutic discoveries for the treatment of uveitis. (AU)

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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)
POSSEBON, LUCAS; COSTA, SARA S.; SOUZA, HELENA R.; AZEVEDO, LUCAS R.; SANT'ANA, MONIELLE; IYOMASA-PILON, MELINA M.; OLIANI, SONIA M.; GIROL, ANA PAULA. Mimetic peptide AC2-26 of annexin A1 as a potential therapeutic agent to treat COPD. International Immunopharmacology, v. 63, p. 270-281, . (15/03359-5)
GIROL, ANA PAULA; ZANON, CAROLINE DE FREITAS; CARUSO, ICARO PUTINHON; COSTA, SARA DE SOUZA; SOUZA, HELENA RIBEIRO; CORNELIO, MARINONIO LOPES; OLIANI, SONIA MARIA. Annexin A1 Mimetic Peptide and Piperlongumine: Anti-Inflammatory Profiles in Endotoxin-Induced Uveitis. CELLS, v. 10, n. 11, . (15/03359-5, 19/19949-7)

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