Chemopreventive effects of FITOPROT against 5-fluorouracil-induced toxicity in HaCaT cells

• Published in: Life sciences (1973) Vol. 193; pp. 300 - 308
• Main Authors: dos Santos Filho, Edvande Xavier, da Silva, Artur Christian Garcia, de Ávila, Renato Ivan, Batista, Aline Carvalho, Marreto, Ricardo Neves, Lima, Eliana Martins, de Oliveira, Cecília Maria Alves, Mendonça, Elismauro Francisco, Valadares, Marize Campos
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Inc 15-01-2018; Elsevier BV
• Subjects: 5-Fluorouracil; Anticarcinogenic Agents - pharmacology; Antioxidants - pharmacology; Bidens pilosa; Cancer; Cell Line; Cell Line, Tumor; Cell Proliferation - drug effects; Cell Survival - drug effects; Cells; Chemoprevention; Curcuma - metabolism; Curcuma - physiology; Curcuma longa; Curcuminoids; Cytochrome; Cytochrome c; Cytokines; Cytokines - metabolism; Cytometry; Cytoplasm; Cytotoxicity; Flavonoids; Flavonoids - pharmacology; Flow cytometry; Fluorescence; Fluorescence microscopy; Fluorouracil - adverse effects; Fluorouracil - pharmacology; Humans; Inflammation; Interleukin 6; Interleukin 8; Interleukin-1beta - pharmacology; Interleukin-6 - pharmacology; Keratinocytes - metabolism; Ligases - metabolism; Ligases - pharmacology; Ligases - therapeutic use; Liquid chromatography; Mass spectrometry; Mass spectroscopy; Mitochondria; Mucositis; NF-kappa B - metabolism; NF-κB protein; Oral mucositis; Oxidative stress; Protective Agents - pharmacology; Proteins; Quercetin; Rutin; Scientific imaging; Stomatitis - drug therapy; Stomatitis - metabolism; Stomatitis - prevention & control; Studies; Toxicity; Tumor necrosis factor; Tumor Necrosis Factor-alpha - pharmacology; Oral mucositis; Curcuma longa; Chemoprevention; Curcuminoids; Bidens pilosa
• ISSN: 0024-3205; 1879-0631
• DOI: 10.1016/j.lfs.2017.09.035

This study evaluated the mechanisms involved in the chemopreventive effects of a mucoadhesive formulation (FITOPROT), containing curcuminoids from Curcuma longa L. (Zingiberaceae) and Bidens pilosa L. (Asteraceae) extract, against 5-FU-induced cellular toxicity using an in vitro oral mucositis model.
Effects of FITOPROT on 5-FU-induced cytotoxicity in HaCaT and SSC-4 cells were evaluated by MTT assay. For mechanistic analyses, HaCaT cells were first pretreated with FITOPROT (0.005%) for 24h followed by treatment with FITOPROT and simultaneously exposed to 5-FU (10μg/mL) for additional 24h.
FITOPROT was able to protect HaCaT cells from 5-FU-triggered cell damage. Moreover, the FITOPROT+5-FU association showed higher cytotoxic effects on SSC-4 cancer cells. Flow cytometry and/or fluorescence microscopy analysis showed FITOPROT was able to significantly reduce ROS generation and prevent mitochondrial changes in HaCaT cells. In addition, it avoided the release of cytochrome c from mitochondria to the cytoplasm in cells exposed to 5-FU, and restored their proliferative activity via Ki-67 expression. Furthermore, FITOPROT regulated 5-FU-induced oxidative stress via Nrf2 involvement. HaCaT cells pretreated/treated with FITOPROT also showed normal expression of TNF-R1 and NF-κB inflammatory proteins and decreased levels of pro-inflammatory cytokines (TNF, IL-1β, IL-6 and IL-8). Moreover, a high-resolution liquid chromatography-mass spectrometry analysis showed the presence of flavonoids rutin, glucoronylated quercetin and dimethylquercetin rutenoside in FITOPROT.
It was showed that FITOPROT, an antioxidant phytochemicals-rich mucoadhesive formulation, exerts chemopreventive effects against 5-FU-triggered toxicity through antioxidant and anti-inflammatory mechanisms and restoration of proliferative capacity in HaCaT cells.