Evaluation of antioxidant, wound healing and anticancer activities of araticum (Annona crassiflora Mart.) in vitro

Author(s)

Lívia Garcia Prado

• Document type: Master's Dissertation
• Press: Campinas, SP.
• Institution: Universidade Estadual de Campinas (UNICAMP). Faculdade de Engenharia de Alimentos
• Defense date: 2018-05-25

Examining board members

Glaucia Maria Pastore; Giovanna Barbarini Longato; Juliano Lemos Bicas

• Advisor: Glaucia Maria Pastore; João Ernesto de Carvalho

Abstract

This study aimed to characterize the crude extracts of araticum peel and seed, the quantification of phenolic compounds, flavonoids and condensed tannins and the analysis of the antioxidant, wound healing and anticancer activities in vitro of these extracts. The characterization of the crude extracts was performed by UHPLC-ESI-MS/MS analysis. The quantification of total phenolic compounds, flavonoids and tannins present in the samples, as well as the antioxidant capacity evaluated by the DPPH and TEAC assays were performed by spectrophotometer analysis and the evaluation of the antioxidant capacity by ORAC was performed in a microplate reader. Antiproliferative assay was performed with eight tumor lines (U251, MCF-7, NCI-ADR/RES, NCI-H460, PC-3, OVCAR-3, HT-29 e K562) and two non-tumoral lines (HaCaT e 3T3) by staining viable proteins with sulforhodamine B. Cell migration experiment was performed with immortalized human keratinocytes (HaCat) and murine fibroblasts (3T3) and the analysis of cell cycle with glioma cells (U251). The compounds epicatechin (6221.63 µg/g) and catechin (579.40 µg/g) were identified as major compounds of the peel extract. This extract also showed 311.03 mg/g phenolic compounds, 117.12 mg/g flavonoids, 179.94 mg/g tannins and antioxidant activity proven by the DPPH (1065 µmolTE/g), TEAC (2022.13 µmolTE/g), hydrophilic ORAC (1302.68 µmolTE/g) and lipophilic ORAC (4643.78 µmolTE/g) assays, respectively. In the seed extract, the major compounds identified were quercetin (21.11 µg/g) and chlorogenic acid (16.41 µg/g). A total of 214.04 mg/g phenolic compounds, 23.74 mg/g flavonoids and 3.27 mg/g tannins was quantified and the antioxidant activity was proven by the DPPH (917 µmolTE/g), TEAC (190.54 µmolTE/g), hydrophilic ORAC (557.10 µmolTE/g) and lipophilic ORAC (5166.31 µmolTE/g) assays. In the antiproliferative assay the crude peel extract presented TGI significant value for glioma (U251 ¿ TGI = 37.64 µg/mL) tumor line. While the crude extract of the seeds presented antiproliferative activity especially in ovary strains with resistance phenotype (NCI-ADR/RES ¿ TGI = 5.36 µg/mL) and prostate (PC-3 ¿ TGI = 16.60 µg/mL). Seed extract arrest the cell cycle in the G1 and G2/M stages of glioma cells (U251), presenting potential as a mitotic inhibitor, while the peel extract arrest the G1 stage. In the migration test performed with keratinocytes (HaCaT), the seed extract induced 73% slot closure at its highest concentration (3.6 µg/mL) and the peel extract inhibited the migration, allowing only 3,8% slot closure at its highest concentration, (36 µg/mL). In the murine fibroblast (3T3) assay, the peel extract induced 34% closure (3.1 µg/mL) and the seed 28% (1.5 µg/mL). Both extracts of araticum presented antioxidant and anticancer activities, and the seed extract presented a healing action. These results suggest a potential in the use of araticum as a source of new drugs or in the enrichment of processed foods (AU)

• FAPESP's process: 16/15746-6 - Evaluation of wound healing properties of araticum (Annona crassiflora Mart.)in vitro
• Grantee: Lívia Garcia Prado
• Support Opportunities: Scholarships in Brazil - Master