Phospholipase D-mediated Activation of IQGAP1 through Rac1 Regulates Hyperoxia-induced p47phoxTranslocation and Reactive Oxygen Species Generation in Lung Endothelial Cells

SummaryLipoxins and their aspirin-triggered carbon-15 epimers have emerged as mediators of key events in endogenous anti-inflammation and resolution. However, the implication of these novel lipid mediators on cardiovascular diseases such as hypertension, atherosclerosis, and heart failure has not been investigated. One of the major features shared by these pathological conditions is the increased production of reactive oxygen species (ROS) generated by vascular NAD(P)H oxidase activation. In this study, we have examined whether an aspirin-triggered lipoxin A4 analog (ATL-1) modulates ROS generation in endothelial cells (EC). Pre-treatment of EC with ATL-1 (1–100 nM) completely blocked ROS production triggered by different agents, as assessed by dihydrorhodamine 123 and hydroethidine. Furthermore, ATL-1 inhibited the phosphorylation and translocation of the cytosplamic NAD(P)H oxidase subunit p47phox to the cell membrane as well as NAD(P)H oxidase activity. Western blot and immunofluorescence microscopy analyses showed that ATL-1 (100 nM) impaired the redox-sensitive activation of the transcriptional factor NF-κB, a critical step in several events associated to vascular pathologies. These results demonstrate that ATL-1 suppresses NAD(P)H oxidase-mediated ROS generation in EC, strongly indicating that lipoxins may play a protective role against the development and progression of cardiovascular diseases.

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Activation of NF-κB induced by H2O2 and TNF-α and its effects on ICAM-1 expression in endothelial cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.2000.279.2.l302 ◽

2000 ◽

Vol 279 (2) ◽

pp. L302-L311 ◽

Cited By ~ 87

Author(s):

Andrea L. True ◽

Arshad Rahman ◽

Asrar B. Malik

Keyword(s):

Gene Expression ◽

Reactive Oxygen Species ◽

Endothelial Cells ◽

Nuclear Dna ◽

Reactive Oxygen Species Generation ◽

Reactive Oxygen ◽

Intercellular Adhesion Molecule ◽

Oxygen Species ◽

Tnf Α

Reactive oxygen species have been proposed to signal the activation of the transcription factor nuclear factor (NF)-κB in response to tumor necrosis factor (TNF)-α challenge. In the present study, we investigated the effects of H2O2 and TNF-α in mediating activation of NF-κB and transcription of the intercellular adhesion molecule (ICAM)-1 gene. Northern blot analysis showed that TNF-α exposure of human dermal microvascular endothelial cells (HMEC-1) induced marked increases in ICAM-1 mRNA and cell surface protein expression. In contrast, H2O2 added at subcytolytic concentrations failed to activate ICAM-1 expression. Challenge with H2O2 also failed to induce NF-κB-driven reporter gene expression in the transduced HMEC-1 cells, whereas TNF-α increased the NF-κB-driven gene expression ∼10-fold. Gel supershift assay revealed the presence of p65 (Rel A), p50, and c-Rel in both H2O2- and TNF-α-induced NF-κB complexes bound to the ICAM-1 promoter, with the binding of the p65 subunit being the most prominent. In vivo phosphorylation studies, however, showed that TNF-α exposure induced marked phosphorylation of NF-κB p65 in HMEC-1 cells, whereas H2O2 had no effect. These results suggest that reactive oxygen species generation in endothelial cells mediates the binding of NF-κB to nuclear DNA, whereas TNF-α generates additional signals that induce phosphorylation of the bound NF-κB p65 and confer transcriptional competency to NF-κB.

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HO-2 provides endogenous protection against oxidative stress and apoptosis caused by TNF-α in cerebral vascular endothelial cells

AJP Cell Physiology ◽

10.1152/ajpcell.00032.2006 ◽

2006 ◽

Vol 291 (5) ◽

pp. C897-C908 ◽

Cited By ~ 75

Author(s):

Shyamali Basuroy ◽

Sujoy Bhattacharya ◽

Dilyara Tcheranova ◽

Yan Qu ◽

Raymond F. Regan ◽

...

Keyword(s):

Oxidative Stress ◽

Reactive Oxygen Species ◽

Endothelial Cells ◽

Reactive Oxygen Species Generation ◽

Reactive Oxygen ◽

Serum Deprivation ◽

Oxygen Species ◽

Tnf Α ◽

Endogenous Protection ◽

Cerebral Vascular

Tumor necrosis factor-α (TNF-α) causes oxidative stress and apoptosis in a variety of cell types. Heme oxygenase (HO) degrades heme to bilirubin, an antioxidant, and carbon monoxide (CO), a cell cycle modulator, and a vasodilator. Newborn pig cerebral microvascular endothelial cells (CMVEC) highly express constitutive HO-2. We investigated the role of HO-2 in protection against TNF-α-induced apoptosis in cerebral vascular endothelium. In CMVEC from mice and newborn pigs, 15 ng/ml TNF-α alone, or with 10 μg/ml cycloheximide (CHX) caused apoptosis detected by nuclear translocation of p65 NF-κB, caspase-3 activation, DNA fragmentation, cell-cell contact destabilization, and cell detachment. TNF-α did not induce HO-1 expression in CMVEC. CMVEC from HO-2 knockout mice showed greater sensitivity to apoptosis caused by serum deprivation and TNF-α than did wild-type mice. TNF-α increased reactive oxygen species generation, including hydrogen peroxide and superoxide radicals, as detected by dihydrorhodamine-123 and dihydroethidium. The TNF-α response was inhibited by superoxide dismutase and catalase suggesting apoptosis is oxidative stress related. Inhibition of endogenous HO-2 in newborn pig CMVEC increased oxidative stress and exaggerated apoptosis caused by serum deprivation and TNF-α. In HO-1-overexpressing CMVEC (HO-1 selective induction by cobalt portophyrin), TNF-α did not cause apoptosis. A CO-releasing compound, CORM-A1, and bilirubin blocked TNF-α-induced reactive oxygen species accumulation and apoptosis consistent with the antioxidant and antiapoptotic roles of the end products of HO activity. We conclude that HO-2 is critical for protection of cerebrovascular endothelium against apoptotic changes induced by oxidative stress and cytokine-mediated inflammation.

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C-peptide reduces high-glucose-induced apoptosis of endothelial cells and decreases NAD(P)H-oxidase reactive oxygen species generation in human aortic endothelial cells

Diabetologia ◽

10.1007/s00125-011-2251-0 ◽

2011 ◽

Vol 54 (10) ◽

pp. 2702-2712 ◽

Cited By ~ 39

Author(s):

V. Cifarelli ◽

X. Geng ◽

A. Styche ◽

R. Lakomy ◽

M. Trucco ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Endothelial Cells ◽

High Glucose ◽

Reactive Oxygen Species Generation ◽

Reactive Oxygen ◽

Oxygen Species ◽

C Peptide ◽

Aortic Endothelial Cells ◽

Induced Apoptosis ◽

Human Aortic Endothelial Cells

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Endothelial Cells Negatively Modulate Reactive Oxygen Species Generation in Vascular Smooth Muscle Cells

Hypertension ◽

10.1161/hypertensionaha.109.133983 ◽

2009 ◽

Vol 54 (2) ◽

pp. 427-433 ◽

Cited By ~ 29

Author(s):

Shaoping Xu ◽

Ying He ◽

Martina Vokurkova ◽

Rhian M. Touyz

Keyword(s):

Reactive Oxygen Species ◽

Endothelial Cells ◽

Smooth Muscle ◽

Vascular Smooth Muscle ◽

Smooth Muscle Cells ◽

Vascular Smooth Muscle Cells ◽

Reactive Oxygen Species Generation ◽

Reactive Oxygen ◽

Muscle Cells ◽

Oxygen Species

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38 NOX4, A HOMOLOGUE OF NOX2, REGULATES HYPEROXIA-INDUCED REACTIVE OXYGEN SPECIES PRODUCTION AND ANGIOGENESIS IN HUMAN LUNG ENDOTHELIAL CELLS.

Journal of Investigative Medicine ◽

10.2310/6650.2005.x0015.37 ◽

2006 ◽

Vol 54 (2) ◽

pp. S349.6-S349

Author(s):

S. Pendyala ◽

I. Gorshkova ◽

B. Gorshko ◽

H. Donghong ◽

R. K. Stern ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Endothelial Cells ◽

Reactive Oxygen Species Production ◽

Human Lung ◽

Reactive Oxygen ◽

Oxygen Species ◽

Lung Endothelial Cells ◽

Species Production

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The Protective Effect of Ubiquinone against the Amyloid Peptide in Endothelial Cells Is Isoprenoid Chain Length-Dependent

Antioxidants ◽

10.3390/antiox10111806 ◽

2021 ◽

Vol 10 (11) ◽

pp. 1806

Author(s):

Javier Frontiñán-Rubio ◽

Yoana Rabanal-Ruiz ◽

Mario Durán-Prado ◽

Francisco Javier Alcain

Keyword(s):

Reactive Oxygen Species ◽

Endothelial Cells ◽

Nadph Oxidase ◽

Protective Effect ◽

Vascular Dysfunction ◽

Reactive Oxygen Species Generation ◽

Density Lipoprotein ◽

Reactive Oxygen ◽

Amyloid Peptide ◽

Oxygen Species

Vascular brain pathology constitutes a common feature in neurodegenerative diseases that could underlie their development. Indeed, vascular dysfunction acts synergistically with neurodegenerative changes to exacerbate the cognitive impairment found in Alzheimer’s disease. Different injuries such as hypertension, high glucose, atherosclerosis associated with oxidized low-density lipoprotein or inflammation induce NADPH oxidase activation, overproduction of reactive oxygen species, and apoptosis in endothelial cells. Since it has been shown that pretreatment of cultured endothelial cells with the lipophilic antioxidant coenzyme Q10 (CoQ10) displays a protective effect against the deleterious injuries caused by different agents, this study explores the cytoprotective role of different CoQs homologues against Aβ25–35-induced damage and demonstrates that only pretreatment with CoQ10 protects endothelial brain cells from Aβ25–35-induced damage. Herein, we show that CoQ10 constitutes the most effective ubiquinone in preventing NADPH oxidase activity and reducing both reactive oxygen species generation and the increase in free cytosolic Ca2+ induced by Aβ25–35, ultimately preventing apoptosis and necrosis. The specific cytoprotective effect of CoQ with a side chain of 10 isoprenoid units could be explained by the fact that CoQ10 is the only ubiquinone that significantly reduces the entry of Aβ25–35 into the mitochondria.

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