Investigation of anti-inflammatory and antioxidant actions of jussara extract (Euterpe edulis Martius) in BV2 cells (microglia) treated with LPS

• Grant number: 19/22071-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Effective date (Start): July 01, 2020
• Effective date (End): June 30, 2021
• Field of knowledge: Health Sciences - Nutrition - Malnutrition and Physiological Development
• Principal Investigator: Cristiano Mendes da Silva
• Grantee: Valkiria Guilherme Assis da Silva
• Host Institution: Instituto de Saúde e Sociedade (ISS). Universidade Federal de São Paulo (UNIFESP). Campus Baixada Santista. Santos , SP, Brazil

Abstract

The induced by high-fat diets (HFDs) obesity promotes a pro-inflammatory state in peripheral and central tissues activating the signaling cascade of the toll-like receptor 4 (TLR4), which induces the production of pro-inflammatory cytokines and may cause metabolic changes, damage and cell death. The TLR4 can be activated through substances of bacterial origin, such as Lipopolysaccharide (LPS) and/or by saturated fatty acids. In this context, the LPS plasma increased due to obesity and/or HFDs promotes: 1) activation of TLR4, which signals positively to c-jun-NH2-kinase (JNK), which stimulates the production of pro-cytokines inflammatory, such as TNF-± which can induce cell death due to apoptosis and 2) increase in free radicals generating a state of oxidative stress inducing damage to cell biomolecules. Therefore, different interventions with anti-inflammatory and antioxidant actions are being studied, such as phenolic compounds, anthocyanin-like found in the fruit of the jussara (Euterpe edulis Martius), revealing a high bioactive potential, but a few studied, particularly in the nervous system central. Aim: To evaluate the responses of BV2 cells (microglia) treated or not with juçara extract, exposed to the pro-inflammatory stimuli induced by LPS. Methodology: BV2 cells will be cultured in DMEM medium containing 10% Bovine Fetal Serum (SFB) at 37°C, under an atmosphere of 5% CO‚. The cytotoxicity test will be performed with different doses of LPS, then the cell treatment will be carried out, associated or not with the extract of Juçara. The cells' supernatant will be used in the ELISA technique to quantify pro-inflammatory (TNF-alpha and IL-1²) and anti-inflammatory (IL-10) cytokines. The cells' protein extract will be analyzed using the Western Blotting technique to evaluate the protein expression of JNK and p-JNK, proteins of the inflammatory pathway. In addition, oxidative stress markers will also be determined by determining the activity of the enzymes Superoxide Dismutase Total, Catalase and Glutathione Peroxidase.