| Grant number: | 16/02021-3 |
| Support Opportunities: | Scholarships in Brazil - Master |
| Start date: | March 01, 2016 |
| End date: | November 30, 2017 |
| Field of knowledge: | Biological Sciences - Physiology - General Physiology |
| Principal Investigator: | Hosana Gomes Rodrigues |
| Grantee: | Thamiris Candreva Robles |
| Host Institution: | Faculdade de Ciências Aplicadas (FCA). Universidade Estadual de Campinas (UNICAMP). Limeira , SP, Brazil |
| Associated research grant: | 13/06810-4 - Mechanisms of action of omega-3 and omega-6 in the tissue repair process: neuro-immune focus, AP.JP |
| Associated scholarship(s): | 16/23298-3 - Analysis of phosphatidylcholine lipid fraction in fat-1 animals in the context of wound healing, BE.EP.MS |
Abstract Inflammation is an essential process in the restoration of local homeostasis and tissue regeneration. In previous studies, we found that FAT-1 animals, which endogenously produce n-3 fatty acids such as EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid), exhibit delayed healing process. These animals have a increase in the activity of myeloperoxidase (MPO), an indirect marker of inflammatory cells migration. At the same time in primary culture of peritoneal macrophages isolated from these animals, production of proinflammatory chemokines and chemokine-derived keratinocytes (KC) and production of reactive oxygen species (ROS), were increased. We also observed a reduction in concentrations of interleukin 10 (IL-10), an anti-inflammatory cytokine. Thus, we believe that the FAT-1 animals are having a proinflammatory behavior. However, so far we do not know which pathway may be modified in these animals. Currently, it has been described that the central nervous system (CNS) interacts with the immune system (IS) through the vagus nerve, which release acetylcholine which will act via alpha-7 nicotinic receptor in cells such as macrophages, inhibiting the production of inflammatory mediators, controlling inflammation. Considering that the healing process is impaired in FAT-1 animals, we will investigate in this project whether this response involves anti-inflammatory cholinergic pathway. For this, we will evaluate the local inflammatory response in the presence / absence of acetylcholine in control animals and FAT-1. We use agonist (PNU) and antagonist (MLA) of ±-7 nicotinic receptor (±-7nAChR), to demonstrate the essential nature of this pathway for the healing process. We will use flow cytometry to assess the inflammatory response (CBA assays) and immunophenotyping, to monitor the process of cell migration during different phases of the healing process; Western blotting to analyze the phosphorylation status of the pathway proteins (JAK-2 and STAT-3) and colorimetry to measure the activity of synthesis enzyme (choline acetyltransferase) and breakdown (acetylcholinesterase) of acetylcholine, the main neurotransmitter via cholinergic . The gene expression analysis will be performed by PCR specific array for wound healing, which evaluate genes directly related to migration processes, production of inflammatory mediators, extracellular matrix formation and remodeling. | |
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