Abstract
The aim of this study is to assess the cytoprotection and potential molecular mechanisms of procyanidin B2 (PCB2) on hydrogen peroxide (H2O2)-induced oxidative damage in MCF-7 cells. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to determine the viability of MCF-7 cell exposure to H2O2 or PCB2. We measured the antioxidant properties of PCB2 by determining the activities of SOD, GSH-Px, LDH and MDA levels, and evaluated apoptosis and intracellular reactive oxygen species (ROS) levels. The related proteins expression levels were monitored by Western blot. MCF-7 cells induced with H2O2 had a remarkable decrease in cell viability that was suppressed when it was interfered with PCB2 (0.1–10.0 μM). PCB2 interference memorably and dose-dependently inhibited H2O2-induced LDH leakage, ROS and MDA overproduction, while PCB2 markedly increased H2O2-induced the activities of SOD and GSH-Px. Eventually, H2O2 prominently down-regulated the ratio of Bcl-2/Bax and the relative proteins expression levels of Nrf2, GCLC, NQO1 and HO-1, and up-regulated the relative proteins expression levels of cytochrome c, caspase-3 and Keap1. However, the relative expression levels of these proteins were reversed in PCB2-interfered MCF-7 cells. This study implied that protective effect of PCB2 on H2O2-induced oxidative damage in MCF-7 cells might be related to inhibition of mitochondria-dependent apoptosis, activation of Keap1/Nrf2/HO-1 signaling pathway and improvement of the antioxidant enzymes activities.
Highlights
Oxidative damage is caused by excessive generation of reactive oxygen species (ROS) in terms of hydroxyl radicals, singlet oxygen and superoxide anions [1]
Determination of the dosage range of P CB2 Cytotoxicity of PCB2 (0.1–40.0 μM) on MCF-7 cells was determined by MTT assay. PCB2 did not show any prominent cytotoxic effect when the concentrations of PCB2 were in the range of 0.1 to 10.0 μM (Fig. 3a)
MCF-7 cells were initially treated with H2O2 (300 μM) for 4 h, and intervented with different concentrations of PCB2 for 24 h
Summary
Oxidative damage is caused by excessive generation of reactive oxygen species (ROS) in terms of hydroxyl radicals, singlet oxygen and superoxide anions [1]. Growing researches have confirmed that oxidative damage is correlated with the pathological development of cancer, atherosclerosis, diabetes, neurological diseases and other diseases [2, 3]. Tan et al Appl Biol Chem (2020) 63:58 breast protective potential, which could scavenge excess free radicals and prevent cell oxidative damage. Many researches have authenticated that PCB2 could regulate the redox state of cells and the protect of antioxidant enzymes in colon cells against oxidative damage and exogenous substances [12, 13]. Up to date, there is not enough information about the effect of PCB2 on breast cells (MCF-7 cells), and the underlying molecular mechanism of the protective effect of PCB2 on hydrogen peroxide (H2O2)-induced oxidative damage remains to be elucidated
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