scholarly journals Intracellular redox state regulates activation of nuclear factor .KAPPA.B and expression of the genes for adhesion molecule in human pulmonary artery endothelial cells.

Ensho ◽  
1999 ◽  
Vol 19 (1) ◽  
pp. 39-46
Author(s):  
Masashi Bando ◽  
Yoshiki Ishii ◽  
Satoshi Kitamura
Keyword(s):  
Endothelial Cells ◽  
Pulmonary Artery ◽  
Adhesion Molecule ◽  
Redox State ◽  
Nuclear Factor ◽  
Human Pulmonary Artery ◽  
Intracellular Redox

EPAS1 (Endothelial PAS Domain Protein 1) Orchestrates Transactivation of Endothelial ICAM1 (Intercellular Adhesion Molecule 1) by Small Nucleolar RNA Host Gene 5 (SNHG5 ) to Promote Hypoxic Pulmonary Hypertension

Hypertension ◽  
2021 ◽  
Author(s):  
Shouxian Wang ◽  
Yu Wang ◽  
Chang Liu ◽  
Gang Xu ◽  
Wenxiang Gao ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Pulmonary Hypertension ◽  
Pulmonary Artery ◽  
Adhesion Molecule ◽  
Intercellular Adhesion Molecule ◽  
Pas Domain ◽  
Intercellular Adhesion Molecule 1 ◽  
Domain Protein ◽  
Human Pulmonary Artery ◽  
Nucleolar Rna

EPAS1 (endothelial PAS domain protein 1), as the major effect gene for the adaptation to chronic hypoxia, is required for hypoxic pulmonary hypertension (HPH). Downregulated EPAS1 ameliorates the development of HPH. We confirmed that EPAS1-specific inhibitor PT2385 ameliorated HPH features, as demonstrated by right ventricle hypertrophy, right ventricular systolic pressure, and pulmonary vascular remodeling. However, the mechanism of EPAS1 in HPH pathogenesis remains unclear. RNA sequencing in human pulmonary artery endothelial cells with EPAS1 knockdown identified EPAS1-regulated genes, including ICAM1 (intercellular adhesion molecule 1), which created a proinflammatory perivascular microenvironment associated with HPH by elevating leukocyte adhesion to the vascular endothelium. Chromatin immunoprecipitation assays revealed that EPAS1 directly bound to ICAM1 promoter. The long noncoding RNA small nucleolar RNA host gene 5 (SNHG5), significantly increased in acute exacerbation period of chronic obstructive pulmonary disease and hypoxic human pulmonary artery endothelial cells, also contributed to the regulation of ICAM1 expression. Endothelial-specific deletion of Snhg5 also rescued HPH in mice. Overexpression of EPAS1 or SNHG5 enhanced, while the depletion of EPAS1 or SNHG5 attenuated, ICAM1 transactivation. SNHG5 was directly regulated by EPAS1, and interestingly, the upregulated SNHG5 could further enhance the levels of EPAS1, which consequently led to hypoxia-induced ICAM1 transactivation. RNA pull-down assay followed by high-throughput sequencing demonstrated that miR-625-5p could bind to SNHG5. Manipulating miR-625-5p altered the levels of EPAS1 during hypoxia. Our data showed a positive feed-forward exists between EPAS1 and SNHG5 signaling during hypoxia-induced ICAM1 transactivation in endothelial cells. Targeting EPAS1 and SNHG5 may provide promising strategies for the prevention of HPH.

scholarly journals Direct Pro-Inflammatory Effect of C-Reactive Protein on Human Pulmonary Artery Endothelial Cells

2012 ◽  
Vol 10 (3) ◽  
pp. 357-363 ◽  
Author(s):  
H.J. Parekh ◽  
H.B. Nguyen ◽  
S.L. Hall ◽  
M. Rehage ◽  
J. Anholm ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Pulmonary Artery ◽  
Adhesion Molecules ◽  
Adhesion Molecule ◽  
Fluorescence Intensity ◽  
Mean Fluorescence Intensity ◽  
Pulmonary Vasculature ◽  
C Reactive Protein ◽  
Reactive Protein ◽  
Human Pulmonary Artery

C-reactive protein (CRP) has a prognostic role in cardiovascular and pulmonary diseases. Recent data suggest its pro-inflammatory effects in atherosclerotic lesion formation. This raises the hypothesis of whether or not CRP has pro-inflammatory effects on pulmonary vasculature by inducing the production of endothelin-1 (ET)-1, a potent vasoconstrictor and proliferative cytokine, and expression of adhesion molecules which could culminate in inflammatory cell recruitment and vascular injury. Human pulmonary artery endothelial cells (HPAECs) were cultured and incubated with 25μg/ml of human recombinant CRP and with interleukin (IL)-1β 10ng/ml, a well-known activator of endothelial cells, which served as a positive control for 24 hours. Expression of vascular cell adhesion molecule (VCAM)-1 and intercellular adhesion molecule (ICAM)-1 was assessed by flow cytometry. Secretion of ET-1 from HPAECs was also evaluated. In this study we show that incubation of HPAECs with human recombinant CRP for 24 hours induced a significant increase in ICAM-1 expression (from 610 to 6553 mean fluorescence intensity, p < 0.005) and VCAM-1 expression (from 212 to 303 mean fluorescence intensity, p < 0.05), as compared to control. Adhesion molecule induction was similar to that observed in endothelial cells activated with IL-1β. Likewise, CRP potentiated the ET-1 production by HPAECs. The levels of ET-1 were significantly higher at 24 hours (control 19.94±3 vs CRP 46.54±18 pg/ml, p < 0.05). In conclusion, this study makes a novel observation that CRP induces expression of adhesion molecules and secretion of ET-1 in HPAECs. Our study provides the first evidence that CRP exerts direct proinflammatory effects on pulmonary artery endothelial cells.

HIV-1 Tat protein-induced VCAM-1 expression in human pulmonary artery endothelial cells and its signaling

2005 ◽  
Vol 289 (2) ◽  
pp. L252-L260 ◽  
Author(s):  
Kai Liu ◽  
David S. Chi ◽  
Chuanfu Li ◽  
H. Kenton Hall ◽  
Denise M. Milhorn ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Pulmonary Artery ◽  
Adhesion Molecule ◽  
Cell Adhesion Molecule ◽  
Ros Generation ◽  
Tat Protein ◽  
Human Pulmonary Artery ◽  
Hiv 1 ◽  
Sb 203580

Expression of cell adhesion molecule in endothelial cells upon activation by human immunodeficiency virus (HIV) infection is associated with the development of atherosclerotic vasculopathy. We postulated that induction of vascular cell adhesion molecule-1 (VCAM-1) by HIV-1 Tat protein in endothelial cells might represent an early event that could culminate in inflammatory cell recruitment and vascular injury. We determined the role of HIV-1 Tat protein in VCAM-1 expression in human pulmonary artery endothelial cells (HPAEC). HIV-1 Tat protein treatment significantly increased cell-surface expression of VCAM-1 in HPAEC. Consistently, mRNA expression of VCAM-1 was also increased by HIV-1 Tat protein as measured by RT-PCR. HIV-1 Tat protein-induced VCAM-1 expression was abolished by the NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) and the p38 MAPK inhibitor SB-203580. Furthermore, HIV-1 Tat protein enhanced DNA binding activity of NF-κB, facilitated nuclear translocation of NF-κB subunit p65, and increased production of reactive oxygen species (ROS). Similarly to VCAM-1 expression, HIV-1 Tat protein-induced NF-κB activation and ROS generation were abrogated by PDTC and SB-203580. These data indicate that HIV-1 Tat protein is able to induce VCAM-1 expression in HPAEC, which may represent a pivotal early molecular event in HIV-induced vascular/pulmonary injury. These data also suggest that the molecular mechanism underlying the HIV-1 Tat protein-induced VCAM-1 expression may involve ROS generation, p38 MAPK activation, and NF-κB translocation, which are the characteristics of pulmonary endothelial cell activation.

scholarly journals Fenfluramine-Induced Gene Dysregulation in Human Pulmonary Artery Smooth Muscle and Endothelial Cells

2011 ◽  
Vol 1 (3) ◽  
pp. 405-418 ◽  
Author(s):  
Weijuan Yao ◽  
Wenbo Mu ◽  
Amy Zeifman ◽  
Michelle Lofti ◽  
Carmelle V. Remillard ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Pulmonary Artery ◽  
Gene Dysregulation ◽  
Pulmonary Artery Smooth Muscle ◽  
Human Pulmonary Artery

Abstract 218: Fibrinolytic Reactivity of Venous Endothelial Cells from Different Vascular Beds

2012 ◽  
Vol 32 (suppl_1) ◽  
Author(s):  
Neil G Kumar ◽  
Elisa Roztocil ◽  
John P Cullen ◽  
David L Gillespie
Keyword(s):  
Endothelial Cells ◽  
Pulmonary Artery ◽  
Cell Types ◽  
Iliac Vein ◽  
Vein Thrombosis ◽  
Cellular Expression ◽  
Artery Embolism ◽  
Vascular Beds ◽  
Human Pulmonary Artery ◽  
Pai 1

Objective: Little is known about the molecular biology of endothelial cells from different venous vascular beds. As a result, our treatment of deep vein thrombosis (DVT) and pulmonary artery embolism (PE) remain identical. PAI-1 and tPA are important regulators of thrombosis and fibrinolysis, while ICAM-1 is known to bind fibrinogen. Here, we aim to investigate differences in fibrinolytic reactivity between human iliac vein endothelial cells (HIVECs) and human pulmonary artery endothelial cells (HPAECs). Methods: Confluent HIVECs and HPAECs, passages 3 - 6, were cultured in the absence or presence of TNFα (10 ng/mL) for 24 hours. Cellular expression of tPA and PAI-1 as analyzed by Western blot analysis and ICAM-1 as analyzed by flow cytometry were compared to controls. Results: Following TNFα stimulation, PAI-1 was upregulated in both HPAECs and HIVECs, however the upregulation observed in HPAECs was approximately 9-fold the increase observed in HIVECs (relative expression: 3.23 ± 0.52 vs 1.26 ± 0.27, n = 3, p < 0.05). While TNFα had no effect on tPA expression in HIVECs, tPA expression in HPAECS was upregulated by 33% (n = 3, p < 0.05). Although TNFα stimulation increased the number of ICAM-1 positive to approximately 100% in both cell types, a 3-fold greater increase in the Mean Fluorescence Intensity (MFI) was observed in HIVECs when compared to HPAECs (relative MFI: 69.28 ± 13.58 vs 21.92 ± 7.22, n = 3, p <0.05). Conclusions: HPAECs and HIVECs react differently in terms of fibrinolytic potential when challenged with a cytokine associated with systemic inflammation, such as in DVT and PE. These findings suggest that endothelial cells from distinct venous vascular beds may differentially regulate the fibrinolytic pathway, thus demonstrating unique properties of the deep veins and the pulmonary artery to respond to thromboembolism.

Isolation and maintenance of human pulmonary artery endothelial cells in culture isolated from transplant donors

1994 ◽  
Vol 267 (4) ◽  
pp. L406-L413 ◽  
Author(s):  
G. A. Visner ◽  
E. D. Staples ◽  
S. E. Chesrown ◽  
E. R. Block ◽  
D. S. Zander ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Pulmonary Artery ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Immunofluorescent Staining ◽  
Northern Analysis ◽  
Pulmonary Vasculature ◽  
Pulmonary Vascular Disease ◽  
Antigen Uptake ◽  
Human Pulmonary Artery

Even though endothelial cells from different locations have similarities, there are potential morphological and functional differences between cells from different vascular regions, as well as between species. Our laboratory is interested in studying the molecular regulation of vasoactive substances in pulmonary vasculature. Therefore, we have developed reproducible methodology to isolate and maintain cultures of human pulmonary artery endothelial cells. The major innovation has been the employment of sections of pulmonary artery from heart transplant donors, from which endothelial cells are isolated. Cell monolayers were identified as endothelial cells by phase-contrast microscopy. Representative dishes of cells were further characterized by indirect immunofluorescent staining for factor VIII antigen, uptake of acetylated low-density lipoprotein, and electron microscopy. These cells were also evaluated for the expression of endothelin-1 (ET-1), a vasoactive 21-amino acid peptide derived from endothelial cells. The cells expressed ET-1 peptide and mRNA as determined by radioimmunoassay and Northern analysis, respectively. We also demonstrated that these cells are useful in transient transfection experiments for potential evaluation of promoter elements. The availability and relevance of these cells provide an important investigative tool for studies on human pulmonary vascular disease.

Effect of hyperoxia on adhesion molecule expression in human endothelial cells and neutrophils

1997 ◽  
Vol 272 (3) ◽  
pp. L418-L425 ◽  
Author(s):  
Y. Suzuki ◽  
T. Aoki ◽  
O. Takeuchi ◽  
K. Nishio ◽  
K. Suzuki ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Adhesion Molecule ◽  
Umbilical Vein ◽  
Oxygen Toxicity ◽  
Intercellular Adhesion Molecule ◽  
Adhesion Molecule Expression ◽  
Neutrophil Accumulation ◽  
Protein Kinase C Inhibitor ◽  
Human Pulmonary Artery ◽  
Umbilical Vein Endothelial Cells

To investigate the pathogenesis of pulmonary oxygen toxicity, we examined the effect of hyperoxia on adhesion molecule expression in cultured human pulmonary artery endothelial cells (HPAEC) and human umbilical vein endothelial cells (HUVEC). Endothelial cell monolayers were exposed to either hyperoxic (90% O(2)-5% CO(2)) or normoxic (21% O(2)-5% CO(2)) conditions for various periods. The level of intercellular adhesion molecule (ICAM)-1 expression had increased in hyperoxia-exposed HPAEC and HUVEC at 48 h (194 +/- 38 and 233 +/- 56%, respectively; P < 0.001) and at 72 h (200 +/- 43 and 223 +/- 52%, respectively; P < 0.001) compared with normoxic conditions. These hyperoxia-induced ICAM-1 expressions were dose dependently attenuated by a protein kinase C inhibitor (H-7). In contrast, the levels of P-selectin and E-selectin expression in HPAEC and HUVEC were unchanged. The levels of ICAM-1 mRNA and the numbers of adherent neutrophils were increased in HPAEC and HUVEC at 48 and 72 h of hyperoxia. On the other hand, hyperoxia caused neutrophil H(2)O(2) production without affecting the level of CD11/CD18 expression. These results suggest that increased ICAM-1 expression in endothelial cells plays an important role in neutrophil accumulation during hyperoxia.

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