Eriocitrin: implications on inflammation pro-resolutive markers

• Grant number: 20/11672-3
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: December 01, 2021
• End date: September 30, 2024
• Field of knowledge: Health Sciences - Dentistry - Periodontology
• Principal Investigator: Luis Carlos Spolidorio
• Grantee: Jhonatan de Souza Carvalho
• Host Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
• Associated scholarship(s): 22/07485-9 - Resolution and post-resolution of inflammation: what role of macrophages in adapted-homeostasis?, BE.EP.DR

Abstract

New knowledge about how inflammation is resolved signals opportunities for the development of new therapeutic approaches complementary or superior to those traditionally used. Eriocitrin (ERIOC), a citrus flavonoid known for its potent anti-inflammatory and antioxidant properties, may be a candidate to act in this process. This proposal aims to lay the foundations for the development of the preventive use of eriocitrin as a strategy for the control of periodontal inflammations. The hypothesis is that food supplementation with eriocitrin modulates the inflammatory immune process, stimulating anti-inflammatory and pro-resolving events. Subproject # 1: evaluate, in vitro, the action of ERIOC on the polarization of macrophages (pro-inflammatory M1 and anti-inflammatory/pro-resolving M2 profile) and the cellular mechanisms involved. For that, the macrophage cell line RAW264.7 will be incubated with LPS from Escherichia coli associated with IFN-³ and simultaneously treated with eriocitrin for 48 hours. qPCR analyzes will be performed to evaluate the expression of M1 and M2 phenotype markers, quantification of M1 and M2 phenotypes by flow cytometry and evaluation of transcription factors STAT1, STAT6 and PPAR-³ by Western blotting. The expression of cytokines IL-1², IL-4, IL-13, TNF-±, GM-CSF, TGF-² and IL-10 will also be evaluated by multiplex immunoassay. The Subproject #2 aims to assess, in vivo, the impact of dietary supplementation with ERIOC on the nature and action of the leukocyte infiltrate during the inflammatory reaction in periodontal tissues, as well as on the generation of chemical mediators, in an experimental protocol of periodontal inflammatory disease induced by LPS injections. Mice (BALB/c) will be treated with a standard diet or diet enriched with ERIOC for 60 days at concentrations of 25 and 50 mg/kg of body weight/day. On the 30th day, the animals will be subjected to bilateral injections of LPS from Escherichia coli into gingival tissues adjacent to the palatal surface of the first maxillary molars, 3x/week for 4 weeks. At the end of the experimental period, the mice will be euthanized and the samples collected for analysis of bone volume, through computerized microtomography; microscopic analysis of the jaws, liver tissue and gastric mucosa; quantification of lipoxin A4 and evaluation of the expression of cytokines IL-1², TNF±, IL-4, IL-5, IL-6, IL-8, IL-10, IL-13, TGF², GM-CSF by multiplex immunoassay; immunoenzymatic assay (ELISA) for the evaluation of leukotriene B4 and prostaglandin E2; immunohistochemistry for marking macrophages (M1 and M2) and the transcription factor PPAR-³ in the jaws. At the end of the study, it is expected to propose strategies to promote the resolution process of inflammatory diseases. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

CARVALHO, JHONATAN DE SOUZA; RAMADAN, DANIA; DE CARVALHO, GABRIEL GARCIA; DE PAIVA GONCALVES, VINICIUS; PELEGRIN, ALVARO FORMOSO; DE ASSIS, RENATA PIRES; BRUNETTI, IGUATEMY LOURENCO; MUSCARA, MARCELO NICOLAS; SPOLIDORIO, DENISE MADALENA; SPOLIDORIO, LUIS CARLOS. Repercussions of Long-Term Naproxen Administration on LPS-Induced Periodontitis in Male Mice. JOURNAL OF PERIODONTAL RESEARCH, v. N/A, p. 13-pg., 2024-11-28. (20/11672-3, 20/16682-7)