Abstract

The in vitro methods currently used to screen bioactive compounds focus on the use of a single model of oxidative stress. However, this simplistic view may lead to conflicting results. The aim of this study was to evaluate the antioxidant properties of two natural extracts (a mix of red wine polyphenols (RWPs) and epigallocatechin gallate (EGCG)) with three models of oxidative stress induced with hydrogen peroxide (H2O2), a mixture of hypoxanthine and xanthine oxidase (HX/XO), or streptozotocin (STZ) in RINm5F beta cells. We employed multiple approaches to validate their potential as therapeutic treatment options, including cell viability, reactive oxygen species production, and antioxidant enzymes expression. All three oxidative stresses induced a decrease in cell viability and an increase in apoptosis, whereas the level of ROS production was variable depending on the type of stress. The highest level of ROS was found for the HX/XO-induced stress, an increase that was reflected by higher expression antioxidant enzymes. Further, both antioxidant compounds presented beneficial effects during oxidative stress, but EGCG appeared to be a more efficient antioxidant. These data indicate that the efficiency of natural antioxidants is dependent on both the nature of the compound and the type of oxidative stress generated.

Highlights

  • Oxidative stress can be defined as an imbalance between proand antioxidants and is often associated with free radical [1] overproduction and/or defective physiological defence mechanisms resulting in the cell being overwhelmed with oxidizing radicals [2]

  • Rat insulinoma clone m5F (RINm5F) [28] cells were purchased from the American Type Culture Collection (ATCC, Manassas, USA)

  • Cells were cultivated in Roswell Park Memorial Institute (RPMI-1640) medium supplemented with 10% fetal bovine serum

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Summary

Introduction

Oxidative stress can be defined as an imbalance between proand antioxidants and is often associated with free radical [1] overproduction and/or defective physiological defence mechanisms resulting in the cell being overwhelmed with oxidizing radicals [2]. High concentrations of ROS can cause lipid peroxidation, protein oxidation or denaturation, nuclear acid oxidation, and many other macromolecular changes that can lead to serious cellular damage [6]. Such ROS-related damage has been identified to occur in numerous diseases, including metabolic syndrome, diabetes, multiple types of cancer, Alzheimer’s disease, and cardiovascular diseases. Enzymatic defence mechanisms [9], such

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