矢车菊素-3-O-葡萄糖苷保护RAW264.7细胞免受过氧化氢诱导的氧化损伤

本研究旨在探究矢车菊素-3-O-葡萄糖苷(cyanidin-3-O-glucoside,C3G)对H2O2诱导RAW264.7细胞氧化损伤的保护作用及其机制.通过噻唑蓝法测定C3G和过氧化氢分别对RAW264.7细胞存活率的影响;采用酶联免疫吸附测定法检测细胞内超氧化物歧化酶(superoxide dismutase,...

Cyanidin-3-O-Glucoside Protects RAW264.7 Cells against Hydrogen Peroxide-Induced Oxidative Damage

The aim of this study is to evaluate the cytoprotection and potential molecular mechanisms of cyanidin-3-O-glucoside (C3G) on hydrogen peroxide (H2O2)-induced oxidative damage in RAW264.7 cells. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was conducted to determine the viability of RAW264.7 cells exposure to H2O2 or C3G. Meanwhile, we measured the antioxidant properties of C3G by determining the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), nitric oxide (NO) release and malondialdehyde (MDA) levels by enzyme-linked immunosorbent assay (ELISA). 2’,7’-dichlorofluorescin diacetate (DCFH-DA) was employed to evaluate the production of intracellular reactive oxygen species (ROS). Finally, the expression levels of related mRNA/protein were evaluated by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. The results showed that the H2O2-induced decrease in the cell viability of RAW264.7 cells was remarkably suppressed C3G (6.25–25.00 μmol/L). C3G significantly inhibited the H2O2-induced of overproduction of intracellular ROS, NO release and MDA levels, but increased the activities of intracellular SOD and GSH-Px (P < 0.05). In addition, the relative mRNA and protein expression levels of Mst1, Mst2 and Keap1 were up-regulated, while the mRNA and protein relative expression levels of Nrf2 and HO-1 were down-regulated in the 400 μmol/L H2O2-treated group when compared to the vehicle-treated group. However, the above changes were reversed by intervention with C3G. C3G could exert a cytoprotective effect possibly by activating the Mst/Nrf2 signaling pathway and improving the activities of antioxidant enzymes.