Abstract 474: Effects of Cigarette Smoke on CD146 Expression and Function in Pulmonary Endothelial Cells

2012 ◽  
Vol 32 (suppl_1) ◽  
Author(s):  
Adelheid Kratzer ◽  
Jonas Salys ◽  
Benjy Gonzalez ◽  
Hong Wei Chu ◽  
Martin Zamora ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cigarette Smoke ◽  
Immunofluorescent Staining ◽  
Microvascular Endothelial Cells ◽  
Pulmonary Microvascular Endothelial Cells ◽  
Pulmonary Endothelial Cells ◽  
Membrane Bound ◽  
Microvascular Endothelial ◽  
Cd146 Expression

Background and Objectives: Cell adhesion molecule CD146 is a transmembrane glycoprotein constitutively expressed in all types of endothelial cells (EC). It exists in two forms: a membrane-anchored form (CD146) and a soluble, extracellular and cleaved form (sCD146). The plasma concentration of sCD146 is modulated in inflammatory diseases that involve endothelial alterations. We investigated the role of endothelial CD146 in cigarette smoke-induced emphysema in vivo and in pulmonary endothelial cells (EC) in vitro . Methods: Sprague Dawley rats exposed to cigarette smoke for 2 months developed significant emphysematous changes (measured by mean linear intercept). Levels of sCD146 were subsequently measured in the circulation as well as in the bronchoalveolar lavage fluid (BALf) via ELISA. In vitro studies were carried out in rat pulmonary microvascular endothelial cells using CSE. Results: CD146 is highly expressed in rat pulmonary microvascular endothelial cells (RPMVEC) and to a much lower extent, in pulmonary macrovascular endothelial cells (RPAEC). Treatment of RPMVEC with cigarette smoke extract (CSE) in vitro resulted in decreased expression of membrane-bound CD146 as well as a reduced gene expression and increased sCD146 levels in the culture medium after 12 hours. Moreover, CSE-induced downregulation of CD146 expression resulted in increased vascular permeability of RPMVEC, as measured by EVANs Blue assay and migration of CFSE-labeled rat alveolar macrophage. Immunofluorescent staining revealed that CSE treatment resulted in translocation of membrane-bound CD146 into the nucleus. Subsequent western blot analysis showed changes in ERK and AKT activation and signaling. Similar results were found upon siRNA silencing of CD146, implicating a role for CD146 in tissue inflammation and integrity. Circulating levels of sCD146 were also elevated in plasma and BALf of patients with COPD and correlated, in part, with the presence of anti-endothelial autoantibodies. Additionally, we found decreased expression of membrane-bound CD146 in lung tissues of COPD patients. Conclusions: Our data suggest that CD146 plays an important role in pulmonary vascular EC function. Moreover, levels of circulating soluble CD146 can be a predictor of vascular endothelial cell injury.

scholarly journals Donor antibodies to HNA-3a implicated in TRALI reactions prime neutrophils and cause PMN-mediated damage to human pulmonary microvascular endothelial cells in a two-event in vitro model

Blood ◽  
2006 ◽  
Vol 109 (4) ◽  
pp. 1752-1755 ◽  
Author(s):  
Christopher C. Silliman ◽  
Brian R. Curtis ◽  
Patricia M. Kopko ◽  
Samina Y. Khan ◽  
Marguerite R. Kelher ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
In Vitro Model ◽  
Microvascular Endothelial Cells ◽  
Pulmonary Microvascular Endothelial Cells ◽  
Biologic Response ◽  
Event Model ◽  
Microvascular Endothelial ◽  
Vitro Model ◽  
Related Mortality

Abstract Transfusion-related acute lung injury (TRALI) is the leading cause of transfusion-related mortality. Antibodies to HNA-3a are commonly implicated in TRALI. We hypothesized that HNA-3a antibodies prime neutrophils (PMNs) and cause PMN-mediated cytotoxicity through a two-event pathogenesis. Isolated HNA-3a+ or HNA-3a− PMNs were incubated with plasma containing HNA-3a antibodies implicated in TRALI, and their ability to prime the oxidase was measured. Human pulmonary microvascular endothelial cells (HMVECs) were activated with endotoxin or buffer, HNA-3a+ or HNA-3a− PMNs were added, and the coculture was incubated with plasma ± antibodies to HNA-3a. PMN-mediated damage was measured by counting viable HMVECs/mm2. Plasma containing HNA-3a antibodies primed the fMLP-activated respiratory burst of HNA-3a+, but not HNA-3a−, PMNs and elicited PMN-mediated damage of LPS-activated HMVECs when HNA-3a+, but not HNA-3a−, PMNs were used. Thus, antibodies to HNA-3a primed PMNs and caused PMN-mediated HMVEC cytotoxicity in a two-event model identical to biologic response modifiers implicated in TRALI.

scholarly journals Prohibitin protects against cigarette smoke extract-induced cell apoptosis in cultured human pulmonary microvascular endothelial cells

2020 ◽  
Author(s):  
Yating Peng ◽  
Zijing Zhou ◽  
Aiyuan Zhou ◽  
JiaXi Duan ◽  
Hong Peng ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cigarette Smoke ◽  
Cell Apoptosis ◽  
Cigarette Smoke Extract ◽  
Microvascular Endothelial Cells ◽  
Cigarette Smoke Exposure ◽  
Dependent Manner ◽  
Pulmonary Microvascular Endothelial Cells ◽  
Mitochondrial Transcription Factor ◽  
Microvascular Endothelial

Prohibitin is an evolutionarily conserved and ubiquitously expressed protein in eukaryocyte. It mediate many important roles in cell survival, apoptosis, autophagy and senescence. In the present study, we aimed to explore the role of prohibitin in cigarette smoke extract (CSE)-induced apoptosis of human pulmonary microvascular endothelial cells (HPMECs). For this purpose, HPMECs were trasfected with prohibitin and challenged with CSE. Our results showed that CSE exposure inhibited prohibitin expression in a dose-dependent manner in HPMECs. Overexpression of prohibitin could protect cell from CSE-induced injury by inhibiting CSE-induced cell apoptosis, inhibiting reactive oxygen species (ROS) production, increase mitochondrial membrane potential, increase the content of mitochondrial transcription factor A (mtTFA), IKKα/β phosphorylation and IκB-α degradation. CSE decreases prohibitin expression in endothelial cells and restoration of prohibitin expression in these cells can protect against the deleterious effects of CSE on mitochondrial and cells. We identified prohibitin is a novel regulator of endothelial cell apoptosis and survival in the context of cigarette smoke exposure.

scholarly journals Cigarette Smoke Extract and Lipopolysaccharides Induce Pyroptosis in Rats Pulmonary Microvascular Endothelial Cells

2022 ◽  
Author(s):  
Qizhi Wang ◽  
Min Liu ◽  
Yu Liu ◽  
Zhen Zhang ◽  
Zhengping Bai
Keyword(s):  
Flow Cytometry ◽  
Endothelial Cells ◽  
Cell Viability ◽  
Cigarette Smoke ◽  
Cigarette Smoke Extract ◽  
Microvascular Endothelial Cells ◽  
Dependent Manner ◽  
Pulmonary Microvascular Endothelial Cells ◽  
Microvascular Endothelial ◽  
Dose Dependent

Abstract Objective: To investigate the effects of cigarette smoke extract (CSE) and lipopolysaccharide (LPS) on the activity and pyroptosis of pulmonary microvascular endothelial cells (PMVECs). Methods: PMVECs were cultured without treatment or with CSE (1%-25%), LPS, or CSE+LPS. Cell viability was detected using the CCK8 method. Apoptosis was evaluated by flow cytometry. Cell morphology was evaluated using optical microscopy. The content of IL-1β and IL-18 was measured by ELISA. Results: CSE decreased cell viability in a dose-dependent manner. The cells in the CSE+LPS group showed the most obvious cytomorphological changes and the highest pyroptosis rate under the microscope. Flow cytometry showed that the CSE and LPS groups showed higher apoptosis rates than the blank group; the apoptotic rate in the CSE+LPS group was even higher (P<0.01). Compared with the bkank group, the levels of IL-18 and IL-1β in the cell supernatant of the CSE, LPS, and CSE+LPS groups increased significantly, with significant differences (P<0.01). There were no differences between the CSE and LPS groups (P>0.05). Compared with the CSE and LPS groups, the CSE+LPS group had higher IL-18 and IL-1β (P<0.01). Conclusion: The effect of CSE on cell viability is dose-dependent. CSE+LPS can induce cell pyroptosis and increase the levels of inflammatory cytokines in PMVECs. These observations demonstrated that pyroptosis caused by CSE and LPS might play an important role in pulmonary vascular remodeling.

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