Fitoquímica, toxicidade e avaliação das atividades anti-inflamatória e antioxidante de Eugenia neonitida Sobral (pitangatuba), uma fruta nativa do Brasil

Author(s)

Josy Goldoni Lazarini

• Document type: Doctoral Thesis
• Press: Piracicaba, SP.
• Institution: Universidade Estadual de Campinas (UNICAMP). Faculdade de Odontologia de Piracicaba
• Defense date: 2020-02-21

Examining board members

Pedro Luiz Rosalen; Ana Paula de Souza; Patricia Correa Dias; Masaharu Ikegaki; Ana Marisa Chudzinski Tavassi

• Advisor: Marcelo Franchin; Pedro Luiz Rosalen

Abstract

Brazilian native fruits (FNB) can be classified as "superfruits" due to their rich polyphenolic composition that can modulate the inflammatory process as well as reduce the production of reactive oxygen and nitrogen species (ROS/RNS). This study determined the polyphenolic composition, evaluated the mechanism of anti-inflammatory action, antioxidant activity and systemic toxicity profile of the extract, fraction (F3) and subfraction (S8) of Eugenia neonitida (Ene). The compounds present in Ene, F3, and S8 were quantified and identified by LC-ESI-QTOF-MS. For the studies of anti-inflammatory activity in vitro, RAW 264.7 macrophages (NF-kB-pLUC gene) transfected were used stimulated with LPS and quantification of inflammatory cytokines by ELISA. For the evaluation of the anti-inflammatory mechanism of action in vivo, neutrophil migration tests were performed in the peritoneal cavity of mice challenged with carrageenan, quantification of inflammatory cytokines in vivo using ELISA, intravital microscopy (rolling and leukocyte adhesion) and expression of adhesion molecule (ICAM-1) by Western Blotting. For antioxidant activity, tests against ROS/RNS (ROO, O2 -, HOCl and NO ) were used. Finally, the systemic toxicity profile was tested in Galleria mellonella larvae. Chemical analysis revealed the presence of hydroxybenzoic acid, flavanoids and ellagitannins for Ene and F3 and derivatives of quercetrin, vanillic acid and cumaric acid for S8. Ene reduced the activation of NF-kB and TNF-'alfa' levels in vitro, however, it did not reduce CXCL2/MIP-2; Ene treated in vivo decreased neutrophil migration as well as TNF-'alfa' and CXCL2/MIP-2 levels. Regarding the fraction, F3 reduced the activation of NF-kB, the levels of TNF-'alfa' and CXCL2 / MIP-2 (in vitro and in vivo) and migration of neutrophils. Finally, S8 reduced activation of NF-kB, levels of TNF-'alfa' and CXCL2 / MIP-2 (in vitro and in vivo) and neutrophil migration confirmed by intravital microscopy and reduced expression of ICAM-1 (adhesion molecule) in endothelium. In general, Ene, F3 and S8 exhibited activities against ROS/RNS and none of the samples induced systemic toxicity in G. mellonella. Thus, Ene, F3 and S8 showed no toxicity and exibited anti-inflammatory and antioxidant activities due to their phenolic composition. E. neonitida is a promising source of bioactive compounds little explored, which can favor sectors such as food industries, pharmaceutical and agribusiness, providing the development of new products for human health (AU)

• FAPESP's process: 16/02926-6 - Phytochemistry and bioprospection of antioxidant and anti-inflammatory activities from unexplored Brasilian native fruits.
• Grantee: Josy Goldoni Lazarini
• Support Opportunities: Scholarships in Brazil - Doctorate