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Involvement of NLRP3/inflammasome on aldosterone-induced endothelial dysfunction

Grant number: 15/24796-4
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): February 01, 2016
Effective date (End): December 31, 2016
Field of knowledge:Biological Sciences - Pharmacology - Cardiorenal Pharmacology
Principal researcher:Thiago Bruder Do Nascimento
Grantee:Isabela Orlandin Pequeno
Home Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil


Aldosterone induces endothelial dysfunction by inflammatory and pro-fibrotic-mechanisms-dependent. Nod-like receptors - NLR are cytoplasmic proteins, which recognize endogen molecules and pathogens. These receptors are expressed in cells from immune system, smooth muscle cells and in endothelium. Aldosterone activates NLRP3, member of NLRs family, in cells from immune system and in the vasculature. However, the role of NLRP3/inflammasome on aldosterone-induced endothelial dysfunction, as well as the mechanisms involved on the NLRP3 activation by aldosterone still are unknown. Aim: In the presente purpose we want to elucidate the role of NLRP3 on aldosterone-induced endotelial dysfunction. We will test the hypothese that NLRP3 activation contributes to aldosterone-induced endotelial dysfunction.Material and methods: Aorta will be isolated from male mice C57BLK6J (C) and NLRP3 knockout mice (NLRp3-/-) with 10-12 weeks of age. Aortic rings, intact endothelium, will be incubated qith aldosterone (0,1uM). Concentration-effect curves (CEC) for acytocholine [(ACh) (0,1 nM-10¼M)] will be performed in aortic rings from C incubated with aldosterone, in presence of vehicle or NLRP3 (MCC950, 1 ¼M). CEC for ACh also will be performed in presence of L-NAME or indomethacin, nitric oxide synthaze (NOS) and cyclooxigenase (COX), respectively. to assess whether NLRP3 activation impairs the endotelial factors (NO and PGs) generation/releasing. NLRP3 activation will be performed by western blot. The total and phosphorylated forms of endotelial NOS (eNOS) and cyclooxigenase, COX-1 and 2, also will b analyzed by western blot. Our results will contribiute for understanding of NLRP3 on endotelial dysfunction induced by aldosterone.

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