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| Author(s): |
Thais Francini Garbieri
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | Bauru. |
| Institution: | Universidade de São Paulo (USP). Faculdade de Odontologia de Bauru (FOB/SDB) |
| Defense date: | 2021-06-01 |
| Examining board members: |
Carlos Ferreira dos Santos;
Flavio Augusto Cardoso de Faria;
Ana Paula Favaro Trombone Garlet;
Pedro de Sousa Gomes;
Ana Carolina Morandini Ramos
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| Advisor: | Carlos Ferreira dos Santos |
| Abstract | |
In addition to the ability to regulate, in a systemic manner, several essential functions of the body, such as those related to blood pressure and cardiovascular homeostasis, the Renin-Angiotensin System (RAS) has been reported as an important immunomodulator of local action in different tissues. Its components can exert pro or anti-inflammatory activities, collaborating with recovery of injured tissue or with the maintenance and evolution of inflammation, causing tissue damage. Fibroblast is the most abundant cellular type of connective tissue, which, among other functions, acts as a sentinel, being able to express several inflammatory mediators in the presence of different stimuli. The hypothesis of the present study was that Angiotensin ll (Ang II), once bound to AT1 receptors in human oral cells, would be able to promote the release inflammatory mediators, helping to establish and/or maintain the local inflammatory process. On the other hand, Ang II, linked to AT2 receptors, would act in the opposite way to the characteristics mentioned about AT1 receptors. Therefore, to better understand the role of Ang II in oral inflammatory diseases, this study aimed to investigate, in vitro, the pro-inflammatory and/or anti-inflammatory potential of Ang ll in primary cultures of human oral fibroblasts, after interaction with AT1 and AT2 receptors under baseline conditions and after pre-stimulation with interleukin 1b (IL1b). Primary cultures of fibroblasts were obtained from explants of gingiva, periodontal ligament and dental pulp (n = 3) and were characterized by morphological analysis and expression of FSP (fibroblast surface protein) by immunofluorescence (IF). Cytotoxicity and cell viability was verified by the AlamaBlueâ assay. The relative gene expression was investigated by Quantitative Polymerase Chain Reaction (qPCR) preceded by Reverse Transcription (RT), after stimuli with Ang II, IL1b and AT1 (valsartan) and AT2 receptor antagonist drugs (PD123319). The immunophenotyping of AT1 and AT2 receptors was verified by flow cytometry in baseline culture situations and after stimulation with IL1b. Stimulus with Ang II for 24h, was not able to modulate the expression of inflammatory mediators in any of the cell types. In shorter times of stimulus, the cells responded to some mediators (P <0.05), however, the blockers were not able to antagonize the observed responses. When the cells received IL1b for 24h before Ang II, and was maintained for another 3, 6 and 24h, there was no additive or synergistic effect between these molecules. Both the AT1 and AT2 receptors were detected by flow cytometry. The results of this study suggest that, in the experimental conditions adopted, the gingival, periodontal ligament and dental pulp fibroblasts did not demonstrate an important role in the establishment and/or maintenance of the local inflammatory process, when stimulated with Ang II. (AU) | |
| FAPESP's process: | 17/19227-6 - Investigation of the modulation of the inflammatory response in human oral and immunological cells by Ang II and Ang 1-7 |
| Grantee: | Thais Francini Garbieri |
| Support Opportunities: | Scholarships in Brazil - Doctorate |