scholarly journals Identification of suitable reference genes for real-time quantitative PCR analysis of hydrogen peroxide-treated human umbilical vein endothelial cells

2016 ◽  
Author(s):  
Tianyi Li ◽  
Hongying Diao ◽  
Lei Zhao ◽  
Yue Xing ◽  
Jichang Zhang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Hydrogen Peroxide ◽  
Endothelial Cells ◽  
Reference Genes ◽  
Umbilical Vein ◽  
Pcr Analysis ◽  
Qrt Pcr ◽  
Stress Injuries ◽  
Umbilical Vein Endothelial Cells

Background. Oxidative stress could induce cell injury in vascular endothelial cells, which is the initial event in the development of atherosclerosis. Although quantitative real-time polymerase chain reaction (qRT-PCR) has been widely used in gene expression studies in oxidative stress injuries, using carefully validated reference genes has not cause sufficient attention in related researches. The objective of this study, therefore, was to select a set of stably expressed reference genes for use in qRT-PCR normalization in oxidative stress injuries in human umbilical vein endothelial cells (HUVECs) induced by hydrogen peroxide (H2O2). Methods. HUVECs were treated with different concentrations of H2O2, geNorm and NormFinder software were conducted to evaluate the expression stabilities of 15 candidate reference genes. The optimal number of reference genes needed for qRT-PCR was determined using geNorm. Results. Using geNorm analysis, we found that five stably expressed reference genes were sufficient for normalization in qRT-PCR analysis in HUVECs treated with H2O2. Genes with the most stable expression according to geNorm were U6 and TFRC, RPLP0, GAPDH and ACTB, and were ALAS1, TFRC, U6, GAPDH, and ACTB according to NormFinder. Discussion. Taken together, our study demonstrated that the expression stability of reference genes may differ according to the statistical program used. U6, TFRC, RPLP0, GAPDH, and ACTB was the optimal set of reference genes for studies on gene expression with qRT-PCR assays in HUVECs under oxidative stress study.

scholarly journals Identification of suitable reference genes for real-time quantitative PCR analysis of hydrogen peroxide-treated human umbilical vein endothelial cells

2016 ◽  
Author(s):  
Tianyi Li ◽  
Hongying Diao ◽  
Lei Zhao ◽  
Yue Xing ◽  
Jichang Zhang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Hydrogen Peroxide ◽  
Endothelial Cells ◽  
Reference Genes ◽  
Umbilical Vein ◽  
Pcr Analysis ◽  
Qrt Pcr ◽  
Stress Injuries ◽  
Umbilical Vein Endothelial Cells

Background. Oxidative stress could induce cell injury in vascular endothelial cells, which is the initial event in the development of atherosclerosis. Although quantitative real-time polymerase chain reaction (qRT-PCR) has been widely used in gene expression studies in oxidative stress injuries, using carefully validated reference genes has not cause sufficient attention in related researches. The objective of this study, therefore, was to select a set of stably expressed reference genes for use in qRT-PCR normalization in oxidative stress injuries in human umbilical vein endothelial cells (HUVECs) induced by hydrogen peroxide (H2O2). Methods. HUVECs were treated with different concentrations of H2O2, geNorm and NormFinder software were conducted to evaluate the expression stabilities of 15 candidate reference genes. The optimal number of reference genes needed for qRT-PCR was determined using geNorm. Results. Using geNorm analysis, we found that five stably expressed reference genes were sufficient for normalization in qRT-PCR analysis in HUVECs treated with H2O2. Genes with the most stable expression according to geNorm were U6 and TFRC, RPLP0, GAPDH and ACTB, and were ALAS1, TFRC, U6, GAPDH, and ACTB according to NormFinder. Discussion. Taken together, our study demonstrated that the expression stability of reference genes may differ according to the statistical program used. U6, TFRC, RPLP0, GAPDH, and ACTB was the optimal set of reference genes for studies on gene expression with qRT-PCR assays in HUVECs under oxidative stress study.

scholarly journals Effects of Nebivolol on Endothelial Gene Expression during Oxidative Stress in Human Umbilical Vein Endothelial Cells

2008 ◽  
Vol 2008 ◽  
pp. 1-6 ◽  
Author(s):  
Ulisse Garbin ◽  
Anna Fratta Pasini ◽  
Chiara Stranieri ◽  
Stefania Manfro ◽  
Chiara Mozzini ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Endothelial Cells ◽  
Beta Blocker ◽  
Umbilical Vein ◽  
Beta Blockers ◽  
Antioxidant Properties ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells ◽  
Progression Of Atherosclerosis

The endothelium plays a key role in the development of atherogenesis and its inflammatory and proliferative status influences the progression of atherosclerosis. The aim of this study is to compare the effects of two beta blockers such as nebivolol and atenolol on gene expression in human umbilical vein endothelial cells (HUVECs) following an oxidant stimulus. HUVECs were incubated with nebivolol or atenolol (10 micromol/L) for 24 hours and oxidative stress was induced by the addition of oxidized (ox)-LDL. Ox-LDL upregulated adhesion molecules (ICAM-1, ICAM-2, ICAM-3, E-selectin, and P-selectin); proteins linked to inflammation (IL-6 and TNFalpha), thrombotic state (tissue factor, PAI-1 and uPA), hypertension such as endothelin-1 (ET-1), and vascular remodeling such as metalloproteinases (MMP-2, MMP-9) and protease inhibitor (TIMP-1). The exposure of HUVECs to nebivolol, but not to atenolol, reduced these genes upregulated by oxidative stress both in terms of protein and RNA expression. The known antioxidant properties of the third generation beta blocker nebivolol seem to account to the observed differences seen when compared to atenolol and support the specific potential protective role of this beta blocker on the expression of a number of genes involved in the initiation and progression of atherosclerosis.

scholarly journals Pretreatment with Astragaloside IV protects human umbilical vein endothelial cells from hydrogen peroxide induced oxidative stress and cell dysfunction via inhibiting eNOS uncoupling and NADPH oxidase – ROS – NF-κB pathway

2016 ◽  
Vol 94 (11) ◽  
pp. 1132-1140 ◽  
Author(s):  
Chonghua Xu ◽  
Futian Tang ◽  
Meili Lu ◽  
Jing Yang ◽  
Ronghui Han ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Nitric Oxide ◽  
Hydrogen Peroxide ◽  
Endothelial Cells ◽  
Nadph Oxidase ◽  
Protein Expression ◽  
Umbilical Vein ◽  
Astragaloside Iv ◽  
Enos Uncoupling ◽  
Umbilical Vein Endothelial Cells

Endothelial cell injury caused by reactive oxygen species (ROS) plays a critical role in the pathogenesis of cardiovascular disorders. Astragaloside IV (AsIV) possesses potent antioxidant properties against oxidative stress through undefined mechanism(s). We sought to investigate whether AsIV protects human umbilical vein endothelial cells (HUVECs) from hydrogen peroxide (H2O2) induced oxidative stress focusing on eNOS uncoupling and the NADPH oxidase – ROS – NF-κB pathway. Compared with HUVECs incubated with H2O2 alone, pretreatment with AsIV significantly increased the viability of HUVECs, which was accompanied with apparent increase in nitric oxide (NO) production and decrease in intracellular superoxide anion production. Furthermore, pretreatment with AsIV increased endothelial nitric oxide synthase (eNOS) dimer/monomer ratio and its critical cofactor tetrahydrobiopterin (BH4) content, decreased Nox4 protein expression (the most abundant Nox isoform in HUVECs), inhibited translocation of NF-κB p65 subunit into nuclear fraction while enhanced the protein expression of IκB-α (the inhibitor of NF-κB p65), reduced the levels of IL-1β, IL-6, and TNF-α in HUVECs medium, and decreased iNOS protein expression. These results suggest that AsIV may protect HUVECs from H2O2-induced oxidative stress via inhibiting NADPH oxidase – ROS – NF-κB pathway and eNOS uncoupling.

Protective effects of peoniflorin against hydrogen peroxide-induced oxidative stress in human umbilical vein endothelial cells

2011 ◽  
Vol 89 (6) ◽  
pp. 445-453 ◽  
Author(s):  
Tao Chen ◽  
Zai-pei Guo ◽  
Xiao-yan Jiao ◽  
Yu-hong Zhang ◽  
Jing-yi Li ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Endothelial Cells ◽  
Oxidative Damage ◽  
Umbilical Vein ◽  
Flow Cytometric Analysis ◽  
Vascular Diseases ◽  
Protective Effects ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells

Peoniflorin (PF), extracted from the root of Paeonia lactiflora Pall., has been reported to have anti-inflammation and antioxidant effects in several animal models. Herein, we investigated the protective effects of PF against hydrogen peroxide (H2O2)-induced oxidative damage in human umbilical vein endothelial cells (HUVECs). HUVECs were treated by H2O2 (240 µmol/L) with or without PF. PF significantly increased the percent cell viability of HUVECs injured by H2O2 using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. By flow cytometric analysis, PF markedly attenuated H2O2-induced apoptosis and intracellular reactive oxygen species production. In addition, PF also displayed a dose-dependent reduction of lactate dehydrogenase leakage, malondialdehyde formation, and caspase-3 proteolytic activities in H2O2-treated cells, which was accompanied with a restoration of the activities of endogenous antioxidants, including total superoxide dismutase and glutathione peroxidase. Finally, Western blot data revealed that H2O2 upregulated phosphorylation of extracellular signal-regulated kinase 1/2 in HUVECs, which was almost completely reversed by PF. Taken together, our data provide the first evidence that PF has a protective ability against oxidative damage in HUVECs. PF may be a candidate medicine for the treatment of vascular diseases associated with oxidative stress.

scholarly journals Piper sarmentosum as an antioxidant on oxidative stress in human umbilical vein endothelial cells induced by hydrogen peroxide

2010 ◽  
Vol 11 (5) ◽  
pp. 357-365 ◽  
Author(s):  
Abdul Hamid Hafizah ◽  
Zakaria Zaiton ◽  
Amom Zulkhairi ◽  
Adenan Mohd Ilham ◽  
Megat Mohd Nordin Nor Anita ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Endothelial Cells ◽  
Umbilical Vein ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells

Proteomic analysis of ginsenoside Re attenuates hydrogen peroxide-induced oxidative stress in human umbilical vein endothelial cells

2016 ◽  
Vol 7 (5) ◽  
pp. 2451-2461 ◽  
Author(s):  
Gui-dong Huang ◽  
Xian-feng Zhong ◽  
Ze-yuan Deng ◽  
Rong Zeng
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Endothelial Cells ◽  
Proteomic Analysis ◽  
Active Component ◽  
Umbilical Vein ◽  
Therapeutic Effects ◽  
Ginsenoside Re ◽  
System P ◽  
Umbilical Vein Endothelial Cells

Ginsenoside Re is an active component in ginseng that has attracted much attention because of its evident therapeutic effects on the cardiovascular system.

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