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Updated: May 9, 2026

High Throughput Screening Assessment of Reactive Oxygen Species (ROS) Generation using Dihydroethidium (DHE) Fluorescence Dye
Published on: January 19, 2024
[Resveratrol protects against hydrogen peroxide induced injury in human umbilical vein endothelial cells]
Yu-ting Wang1, Hai-yan Zhou, Xiao-cui Lü
1Department of Anesthesia, Sir Run Run Shaw Hospital, Zhejiang University, Hangzhou 310016, China.
Objective:
To investigate the effect of Resveratrol on Hydrogen peroxide-treated human umbilical vein endothelial cells (HUVEC).
Methods:
Oxidative stress in HUVEC was induced by hydrogen peroxide (H2O2). The cells were divided in to 5 groups: A. Control group; B. H2O2 group; C. Res + H2O2 group; D. Res + H2O2 + LY294002 group; E. H2O2 + LY294002 group. The cell viability were measured by cck-8 assay; Malondialdehyde (MDA) and Superoxide dismutase (SOD) were determined by using commercially available kits; Reactive oxygen species (ROS)were detect by flow cytometer; the expression of Nuclear factor-E2-related factor(Nrf2), Protein kinase B(Akt), Mammalian target of rapamycin (mTOR) were determined by Western blot.
Results:
Compared with control, resveratrol increased the cells viability (36.14 ± 2.20) to (72.05 ± 2.46) (P = 0.004), decreased MDA formation (299.38 ± 12.42) to (159.17 ± 7.30)(P = 0.006), elevated SOD activity(35.61 ± 3.26) to (81.37 ± 4.55) (P = 0.007), inhibited ROS production (177.86 ± 8.98) to (133.96 ± 4.17) (P = 0.003). These changes were accompany with the activation of p-Nrf2 (198.57 ± 6.89) to (371.10 ± 8.41) (P = 0.005), p-Akt (102.93 ± 3.29) to (221.62 ± 6.51) (P = 0.004), p-mTOR (93.26 ± 4.12) to (172.14 ± 5.74) (P = 0.008). However these protective effect of resveratrol were abolished by LY294002.
Conclusion:
Resvertrol could protects HUVEC cells from H2O2-induced oxidative stress, in which the PI3K-Akt-mTOR pathway may be involved.
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