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Investigation of the modulation of the inflammatory response in human oral and immunological cells by Ang II and Ang 1-7

Grant number: 17/19227-6
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): December 01, 2017
Effective date (End): July 31, 2020
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Carlos Ferreira dos Santos
Grantee:Thais Francini Garbieri
Home Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil
Associated research grant:15/03965-2 - Role of the renin-angiotensin system in different oral inflammatory models: an experimental interdisciplinary and clinical approach, AP.TEM
Associated scholarship(s):18/23934-2 - Role of Angiotensin II and angiotensin 1-7 on the metabolism of human bone cells, BE.EP.DR

Abstract

In addition to the ability to regulate various physiological functions such as those related to blood pressure and cardiovascular system, the Renin-Angiotensin System (RAS) has been reported as an important modulator of inflammatory and immunological processes. Its components can exert pro or anti-inflammatory activities, collaborating with the recovery of the damaged tissue or with the maintenance and evolution of inflammation causing tissue damages. The hypothesis of the present study is that Angiotensin II (Ang II) plays an important role in the development of the inflammatory process in the fibroblasts of oral tissues, while Angiotensin 1-7 (Ang 1-7) may attenuate the inflammation. Therefore, in order to better understand the role of RAS components related to oral inflammatory diseases such as periodontitis and pulpitis, the objective of this study is to verify the proinflammatory potential of Ang II and the anti-inflammatory potential of Ang 1-7 in fibroblasts of oral tissues from human gingiva (HGF), human periodontal ligament (HPLF), and human permanent tooth pulp (HPF), in addition, to verify these same parameters in human immune cells, also involved in the inflammatory process. Another objective of this study is to evaluate whether the inflammatory response mediated by Ang II is preceded by activation of the nuclear factor kappa B complex (NF-kB). For this, primary cultures of oral fibroblasts and human immune cells will be obtained from three healthy individuals. The immunofluorescence technique will be performed to characterize the cell types studied. All cell types will be stimulated with Ang II and Ang 1-7, after which cytotoxicity and cell viability will be assessed by means of the MTT assay. The expression of the mRNA of the genes related to the establishment and maintenance of the inflammatory process, as well as the NF-kB complex, will be verified by the Quantitative Polymerase Chain Reaction (qPCR) preceded by Reverse Transcription (RT). The production of the enzymes and cytokines, produced by the different cell types, related to the pathways described, will be evaluated by ELISA (Multiplex). (AU)