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

[1]	[1]LANKIN V Z, TIKHAZE A K.Role of oxidative stress in the genesis of atherosclerosis and diabetes mellitus:a personal look back on 50 years of research[J].Curr Aging Sci, 2017, 10 (1) :18-25.
[2]	[2]PETROU A L, TERZIDAKI A.A meta-analysis and review examining a possible role for oxidative stress and singlet oxygen in diverse diseases[J].Biochem J, 2017, 474 (16) :2713-2731.
[3]	[3]SIES H.Hydrogen peroxide as a central redox signaling molecule in physiological oxidative stress:Oxidative eustress[J].Redox Biol, 2017, 11 (C) :613-619.
[4]	[4]DA ROSA ARAUJO A S, SILVA DE MIRANDA M F, DE OLIVEIRA U O, et al.Increased resistance to hydrogen peroxide-induced cardiac contracture is associated with decreased myocardial oxidative stress in hypothyroid rats[J].Cell Biochem Funct, 2010, 28 (1) :38-44.
[5]	[5]AL-SHEDDI E S, FARSHORI N N, AL-OQAIL M M, et al.Protective effect of lepidium sativum seed extract against hydrogen peroxide-induced cytotoxicity and oxidative stress in human liver cells (Hep G2) [J].Pharm Biol, 2016, 54 (2) :314-321.
[6]	[6]LIU G, LI Y, GAO X G.Micro RNA-181a is upregulated in human atherosclerosis plaques and involves in the oxidative stress-induced endothelial cell dysfunction through direct targeting Bcl-2[J].Eur Rev Med Pharmacol Sci, 2016, 20 (14) :3092-3100.
[7]	[7]NOWAK W N, DENG J, RUAN X Z, et al.Reactive oxygen species generation and atherosclerosis[J].Arterioscler Thromb Vasc Biol, 2017, 37 (5) :e41-e52.
[8]	[8]YANG X, Li Y.Oxidative stress-mediated atherosclerosis:mechanisms and therapies[J].Front Physiol, 2017, 23 (8) :600.
[9]	[9]DAS P, BISWAS S, MUKHERJEE S, et al.Association of oxidative stress and obesity with insulin resistance in type 2diabetes mellitus[J].Mymensingh Med J, 2016, 25 (1) :148-152.
[10]	[10]KWAK H J, CHOI H E, CHEON H G.5-LO inhibition ameliorates palmitic acid-induced ER stress, oxidative stress and insulin resistance via AMPK activation in murine myotubes[J].Sci Rep, 2017, 7 (1) :5025.
[11]	[11]LATHA R, SHANTHI P, SACHDANANDAMP.Kalpaamruthaa modulates oxidative stress in cardiovascular complication associated with type 2 diabetes mellitus through PKC-β/Akt signaling[J].Can J Physiol Pharmacol, 2013, 91 (11) :901-912.
[12]	[12]ASMAT U, ABAD K, ISMAIL K.Diabetes mellitus and oxidative stress-A concise review[J].Saudi Pharm J, 2016, 24 (5) :547-553.
[13]	[13]SIES H, BERNDT C, JONES DP.Oxidative stress[J].Annu Rev Biochem.2017, 86 (20) :715-748.
[14]	[14]XU C, TANG F, LU M, et al.Pretreatment with astragaloside IV protects human umbilical vein endothelial cells from hydrogen peroxide induced oxidative stress and cell dysfunction via inhibiting e NOS uncoupling and NADPH oxidase-ROS-NF-κB pathway[J].Can J Physiol Pharmacol, 2016, 30 (5) :1-9.
[15]	[15]LIN XL, LIU Y, LIU M, et al.Inhibition of hydrogen peroxide-induced human umbilical vein endothelial cells aging by allicin depends on sirtuin1 activation[J].Med Sci Monit, 2017, 23 (31) :563-570.
[16]	[16]CHEN J, GU Y, SHAO Z, et al.Propofol protects against hydrogen peroxide-induced oxidative stress and cell dysfunction in human umbilical vein endothelial cells[J].Mol Cell Biochem, 2010, 339 (1-2) :43-54.

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