CARS microscopy analysis of the interaction of flavonoids (quercetin, rutin and myricetin) and neutrophil extracellular traps (NETs) on A549 cells

• Grant number: 19/01681-8
• Support Opportunities: Scholarships abroad - Research Internship - Scientific Initiation
• Start date: May 01, 2019
• End date: July 31, 2019
• Field of knowledge: Biological Sciences - Immunology - Cellular Immunology
• Principal Investigator: Karina Alves de Toledo
• Grantee: Sarah Mendes
• Supervisor: Jonathan Richard Brewer
• Host Institution: Faculdade de Ciências e Letras (FCL-ASSIS). Universidade Estadual Paulista (UNESP). Campus de Assis. Assis , SP, Brazil
• Institution abroad: University of Southern Denmark (SDU), Denmark
• Associated to the scholarship: 17/05233-4 - Antiinflammatory evaluation of miricetin flavonoid on PMA-activated neutrophils, BP.IC

Abstract

Neutrophils Extracellular Traps (NETs) are released from activated neutrophils during inflammatory process. They capture and/or kill pathogens but they also are cytotoxic to pulmonary cells. Histones, elastase e myeloperoxidase are present in the NETs and they have been described as responsible proteins for the tissue damage induced by NETs. Our preliminary results demonstrated that flavonoids quercetin, rutin and myricetin inhibit the enzymatic role of elastase and myeloperoxidase. In Brazil, we are studying the protective effects of those flavonoids in A549 cells culture maintained in the presence of NETs. During the exchange in Denmark, we will be interested to know how these molecules interact on A549 surface. For this purpose, A549 cells will be incubated with different concentrations of flavonoids (quercetin, rutin and myricetin) and/or NETs (Neutrophil Extracellular Traps). The interaction of the molecules on cell surface will be analyzed by advanced microscopy technique Coherent Anti-Stokes Raman Scattering (CARS). The project will be conducted in the University of Southern Denmark, under supervision of Dr. Jonathan R. Brewer, at the Department of Biochemistry and Molecular Biology at the Danish Molecular Biomedical Imaging Center (DaMBIC). The presence of flavonoids (quercetin, rutin and myricetin) and/or NETs (Neutrophil Extracellular Traps) as such as their distribution on cells surface will be useful to understand the flavonoids protective effects on cell damage induced by NETs. (AU)