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Reticulum stress involvement in fatty acids-induced neutrophils death

Grant number: 12/15079-9
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): March 01, 2013
Effective date (End): December 31, 2014
Field of knowledge:Biological Sciences - Physiology
Principal Investigator:Adriana Cristina Levada-Pires
Grantee:Raquel Freitas Zambonatto
Home Institution: Pró-Reitoria de Pós-Graduação e Pesquisa. Universidade Cruzeiro do Sul (UNICSUL). São Paulo , SP, Brazil
Associated research grant:10/50848-8 - Role of oxygen reactive species and ceramides in human neutrophil death induced by fatty acids, AP.JP

Abstract

The mechanisms involved in neutrophil apoptosis induced by fatty acids and the involvement of reticulum stress in this process will be investigated. Will be investigated the effect of monoynsaturated fatty acid oleic (main component of olive oil)and the polyunsaturated fatty acid n-6 linoleic (main component of soybean oil) and is very abundant in the diet. Human neutrophils will be pre-treated for 3 hours with concentration, non-toxic (100 µM) and toxic (250 µM) of oleic and linoleic acids. The mechanisms involved in AG induced neutrophils death, will be assessed by fluorometry and western blotting. The effect fatty acids on the expression of pro-(Bax) and antiapoptotic (Mcl-1)proteins. The extrinsic (Bid) and intrinsic pathways (cytochrome c) and / or endoplasmic reticulum stress (CHOP, GRP78, PERK, pPERK, peIF2-alpha and caspase 12) involved in neutriphils death also will be determined. To establish the possible involvement of reticulum stress in the death of neutrophils, cells are cultivated with the fatty phenylbutyric acid (4-PBA - inhibitor reticulum stress), and then will be determined in neutrophils the viability and DNA fragmentation by cytometric flow.