Establishment of Oxidative Stress Model by Inducing Human Umbilical Vein Endothelial Cells Using Hydrogen Peroxide

Dai Qing Li; Sun Gui Hu; Yan Bin; Guo Tao; Dai Qing Yuan

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Dai Qing Li , Sun Gui Hu , Yan Bin , Guo Tao , Dai Qing Yuan . Establishment of Oxidative Stress Model by Inducing Human Umbilical Vein Endothelial Cells Using Hydrogen Peroxide[J]. Journal of Kunming Medical University, 2018, 39(04): 34-39.

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Dai Qing Li , Sun Gui Hu , Yan Bin , Guo Tao , Dai Qing Yuan . Establishment of Oxidative Stress Model by Inducing Human Umbilical Vein Endothelial Cells Using Hydrogen Peroxide[J]. Journal of Kunming Medical University, 2018, 39(04): 34-39.

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Establishment of Oxidative Stress Model by Inducing Human Umbilical Vein Endothelial Cells Using Hydrogen Peroxide

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基金：云南省教育厅科学研究基金资助项目 (2015Y153);

Received Date: 2018-02-04

Abstract

Abstract

Objective To establish an oxidative stress cell model by inducing human umbilical vein endothelial cells (HUVECs) using hydrogen peroxide (H2 O2) . Methods Six gradient concentrations of H2 O2 were co-cultured with HUVECs for 6 h, 12 h, 24 h and 48 h. Inverted microscope was used to observe the change of cell morphologies. The optimal working concentration and effect time of H2 O2 on HUVECs were screened by CCK-8 method for cell viability, flowcymetry for apoptosis rate, and CDFU-DA fluorescent probing and flowcymetry for ROS levels. Re s ults Any concentration of H2 O2 could do harm to HUVECs co-culturing with cells. Cell viabilities decreased statistically when treating cells with 400, 600 or 800 μmol/L H2 O2 for different times (P<0.05) . Cell viabilities also decreased gradually as time prolonged when treating cells with 800 μmol/L H2 O2 (P<0.001) . When treating cells with different concentration of H2 O2 for 24 h, cell apoptosis rates increased dramatically (1000μmol/L as an exception, P<0.05) , so did cell necrosis rates (100 μmol/L as an exception, P<0.05) ;When treating cells with 800 μmol/L H2 O2, cell apoptosis rates increased dramatically as time prolonged, reaching the peak at 24 h (P<0.05) , so did necrosis rates, reaching the peak at 48 h (P<0.05) . ROS level reached thepeak while treating cells with 400 μmol/L H2 O2 (P<0.001) ; ROS levels increased gradully as time prolonged when treating cells with 800 μmol/L H2 O2 (P<0.001) . Conclusions The oxidative stress model in HUVECs was established by coculturing cells with culture medium containing 800 μmol/L hydrogen peroxide for 24 h.
- Human umbilical vein endothelial cells,
- Hydrogen peroxide,
- Oxidative stress,
- Cell model

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References(16)

References

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