Phospholipase A2 enzymatic activity as water quality biomarker

• Grant number: 21/03352-1
• Support type: Scholarships in Brazil - Scientific Initiation
• Effective date (Start): June 01, 2021
• Effective date (End): November 30, 2021
• Field of knowledge: Engineering - Sanitary Engineering - Water Resources
• Principal researcher: Bruno Paes de Carli
• Grantee: Isabelly Annunciato
• Home Institution: Instituto de Ciências da Saúde (ICS). Universidade Paulista (UNIP). São Paulo , SP, Brazil

Abstract

Phospholipases A2 are a large family of enzymes that have the function of hydrolyzing the sn2 phosphodiester bond of phospholipids and releasing fatty acids, emphasizing unsaturated fatty acids of 18 to 20 carbons (PUFAs, w-3 and w-6) and acid arachidonic (C20: 4 w-6), whose subsequent metabolization by cyclooxygenase 2 (COX-2) generates bioactive pro-inflammatory and homeostatic lipids. Within the current context, with a pandemic caused by Covid-19, there was an increase in the concentration of drugs used in the early treatment of the disease (ivermectin and chloroquine) in the water of rivers and lakes. The situation worsens in the face of an epidemic of dengue and Chikungunya that has been intensifying in the country, making it necessary to nebulize with organophosphates (malathion) on a recurring basis. PLA2s has the function of acting as a bridge between hormones, inflammatory stimuli and signs of physiological stress that rely on arachidonic acid mobilization to mediate adaptive and physiological responses. Zebrafish is a largest model for biological research due to DNA homology. Thus, in this project we intend to investigate the presence and function of phospholipase A2 enzymes in zebrafish, as well as their modulation in the presence of xenobiotic agents present in freshwater. So, this project will be divided into two parts, the first will be focused on phospholipase A2 activity, in which we will develop a simple method of general and specific evaluation for PLA2 using colorimetric techniques; we will also develop methods or analysis protocols for secretory PLA2 (sPLA2) and cytosolic PLA2 (cPLA2) using specific inhibitors, we will build a in sílico model of zebrafish cPLA2 to evaluate, through molecular modeling, the interaction of compounds with fish cPLA2 and the effect of xenobiotic compounds: chloroquine, ivermectin and malathion on the enzymatic activity of Danio rerio's PLA2. (AU)