scholarly journals AM966, an Antagonist of Lysophosphatidic Acid Receptor 1, Increases Lung Microvascular Endothelial Permeability through Activation of Rho Signaling Pathway and Phosphorylation of VE-Cadherin

2017 ◽  
Vol 2017 ◽  
pp. 1-12 ◽  
Author(s):  
Junting Cai ◽  
Jianxin Wei ◽  
Shuang Li ◽  
Tomeka Suber ◽  
Jing Zhao
Keyword(s):  
Endothelial Cells ◽  
Lung Injury ◽  
Lysophosphatidic Acid ◽  
Endothelial Permeability ◽  
Endothelial Barrier ◽  
Dependent Manner ◽  
Gap Formation ◽  
Barrier Dysfunction ◽  
Lpa Receptor ◽  
Microvascular Endothelial

Maintenance of pulmonary endothelial barrier integrity is important for reducing severity of lung injury. Lysophosphatidic acid (LPA) regulates cell motility, cytoskeletal rearrangement, and cell growth. Knockdown of LPA receptor 1 (LPA1) has been shown to mitigate lung injury and pulmonary fibrosis. AM966, an LPA1 antagonist exhibiting an antifibrotic property, has been considered to be a future antifibrotic medicine. Here, we report an unexpected effect of AM966, which increases lung endothelial barrier permeability. An electric cell-substrate sensing (ECIS) system was used to measure permeability in human lung microvascular endothelial cells (HLMVECs). AM966 decreased the transendothelial electrical resistance (TEER) value immediately in a dose-dependent manner. VE-cadherin and f-actin double immunostaining reveals that AM966 increases stress fibers and gap formation between endothelial cells. AM966 induced phosphorylation of myosin light chain (MLC) through activation of RhoA/Rho kinase pathway. Unlike LPA treatment, AM966 had no effect on phosphorylation of extracellular signal-regulated kinases (Erk). Further, in LPA1 silencing cells, we observed that AM966-increased lung endothelial permeability as well as phosphorylation of VE-cadherin and focal adhesion kinase (FAK) were attenuated. This study reveals that AM966 induces lung endothelial barrier dysfunction, which is regulated by LPA1-mediated activation of RhoA/MLC and phosphorylation of VE-cadherin.

ID: 114: AN ANTAGONIST OF LYSOPHOSPHATIDIC ACID RECEPTOR1, AM966, INCREASES PULMONARY ENDOTHELIAL PERMEABILITY THROUGH ACTIVATION OF VE-CADHERIN

2016 ◽  
Vol 64 (4) ◽  
pp. 965.3-966
Author(s):  
J Cai ◽  
J Wei ◽  
AM Jacko ◽  
J Zhao
Keyword(s):  
Lysophosphatidic Acid ◽  
Barrier Function ◽  
Endothelial Permeability ◽  
Heart Association ◽  
Endothelial Barrier ◽  
Dependent Manner ◽  
Gap Formation ◽  
Endothelial Barrier Function ◽  
Barrier Dysfunction ◽  
Barrier Integrity

BackgroundMaintenance of pulmonary endothelial barrier integrity is important for reducing severity of lung injury. VE-cadherin is a major component of cell–cell adherens junctions in endothelium. In response to inflammatory stimuli, VE-cadherin is tyrosine phosphorylated, resulting in dissociation with catenins, which links to f-actin. Lysophosphatidic acid (LPA) is a bioactive lysophospholipid, which regulates cell motility. LPA has been shown to increase lung epithelial barrier integrity, while it reduces endothelial barrier function. AM966 is an antagonist exhibiting an anti-fibrotic property. However, the effect of AM966 on pulmonary endothelial barrier integrity has not been well studied.Methods and ResultsTo investigate endothelial barrier integrity, electric cell-substrate sensing (ECIS) system was used to measure permeability in human lung microvascular endothelial cells (HLMVECs). Similar to the effect of LPA, AM966 increases permeability immediately in a dose dependent manner. To investigate the molecular mechanism by which regulates AM966-mediated reduction of endothelial barrier function, HLMVECs were treated with AM966, and then phosphorylation of myosin light chain (MLC) and VE-cadherin were determined by immunoblotting. AM966 increased phosphorylation of MLC and VE-cadherin. VE-cadherin and f-actin double immunostaining revealed that AM966 induces gap formation and f-actin stress fibers as well as dissociation between VE-cadherin and f-actin.ConclusionThis study reveals that AM966 induces lung endothelial barrier dysfunction, which is regulated by phosphorylation of VE-cadherin.This work was supported by the National Institutes of Health (R01GM115389 to J.Z.), American Heart Association 12SDG9050005 (J.Z.), American Lung Association Biomedical Research Grant RG350146 (J.Z.).

scholarly journals The HSP90 Inhibitor, AUY-922, Protects and Repairs Human Lung Microvascular Endothelial Cells from Hydrochloric Acid-Induced Endothelial Barrier Dysfunction

Cells ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1489
Author(s):  
Ruben M. L. Colunga Biancatelli ◽  
Pavel Solopov ◽  
Betsy Gregory ◽  
John D. Catravas
Keyword(s):  
Endothelial Cells ◽  
Hydrochloric Acid ◽  
Human Lung ◽  
Hsp90 Inhibitor ◽  
Endothelial Barrier ◽  
Microvascular Endothelial Cells ◽  
Dependent Manner ◽  
Barrier Dysfunction ◽  
Endothelial Barrier Dysfunction ◽  
Microvascular Endothelial

Exposure to hydrochloric acid (HCl) leads acutely to asthma-like symptoms, acute respiratory distress syndrome (ARDS), including compromised alveolo-capillary barrier, and respiratory failure. To better understand the direct effects of HCl on pulmonary endothelial function, we studied the characteristics of HCl-induced endothelial barrier dysfunction in primary cultures of human lung microvascular endothelial cells (HLMVEC), defined the involved molecular pathways, and tested the potentially beneficial effects of Heat Shock Protein 90 (HSP90) inhibitors. HCl impaired barrier function in a time- and concentration-dependent manner and was associated with activation of Protein Kinase B (AKT), Ras homolog family member A (RhoA) and myosin light chain 2 (MLC2), as well as loss of plasmalemmal VE-cadherin, rearrangement of cortical actin, and appearance of inter-endothelial gaps. Pre-treatment or post-treatment of HLMVEC with AUY-922, a third-generation HSP90 inhibitor, prevented and restored HCl-induced endothelial barrier dysfunction. AUY-922 increased the expression of HSP70 and inhibited the activation (phosphorylation) of extracellular-signal regulated kinase (ERK) and AKT. AUY-922 also prevented the HCl-induced activation of RhoA and MLC2 and the internalization of plasmalemmal VE-cadherin. We conclude that, by increasing the expression of cytoprotective proteins, interfering with actomyosin contractility, and enhancing the expression of junction proteins, inhibition of HSP90 may represent a useful approach for the management of HCl-induced endothelial dysfunction and acute lung injury.

scholarly journals Magnitude-dependent effects of cyclic stretch on HGF- and VEGF-induced pulmonary endothelial remodeling and barrier regulation

2008 ◽  
Vol 295 (4) ◽  
pp. L612-L623 ◽  
Author(s):  
Anna A. Birukova ◽  
Nurgul Moldobaeva ◽  
Junjie Xing ◽  
Konstantin G. Birukov
Keyword(s):  
Growth Factor ◽  
Lung Injury ◽  
Synergistic Effects ◽  
Endothelial Permeability ◽  
Small Gtpase ◽  
Cyclic Stretch ◽  
Dependent Manner ◽  
Protective Effects ◽  
Gap Formation ◽  
Barrier Dysfunction

Mechanical ventilation at high tidal volumes compromises the blood-gas barrier and increases lung vascular permeability, which may lead to ventilator-induced lung injury and pulmonary edema. Using pulmonary endothelial cell (ECs) exposed to physiologically [5% cyclic stretch (CS)] and pathologically (18% CS) relevant magnitudes of CS, we evaluated the potential protective effects of hepatocyte growth factor (HGF) on EC barrier dysfunction induced by CS and vascular endothelial growth factor (VEGF). In static culture, HGF enhanced EC barrier function in a Rac-dependent manner and attenuated VEGF-induced EC permeability and paracellular gap formation. The protective effects of HGF were associated with the suppression of Rho-dependent signaling triggered by VEGF. Five percent CS promoted HGF-induced enhancement of the cortical F-actin rim and activation of Rac-dependent signaling, suggesting synergistic barrier-protective effects of physiological CS and HGF. In contrast, 18% CS further enhanced VEGF-induced EC permeability, activation of Rho signaling, and formation of actin stress fibers and paracellular gaps. These effects were attenuated by HGF pretreatment. EC preconditioning at 5% CS before HGF and VEGF further promoted EC barrier maintenance. Our data suggest synergistic effects of HGF and physiological CS in the Rac-mediated mechanisms of EC barrier protection. In turn, HGF reduced the barrier-disruptive effects of VEGF and pathological CS via downregulation of the Rho pathway. These results support the importance of HGF-VEGF balance in control of acute lung injury/acute respiratory distress syndrome severity via small GTPase-dependent regulation of lung endothelial permeability.

Abstract 45: Endothelial Aryl Hydrocarbon Receptor Nucleatranslator (Arnt) is a Novel Regulator of Cardiac Endothelial Barrier Integrity in Diabetes

2017 ◽  
Vol 37 (suppl_1) ◽  
Author(s):  
Maura Knapp ◽  
Mei Zheng ◽  
Nikola Sladojevic ◽  
Qiong Zhao ◽  
Konstaintin G Birukov ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Barrier Function ◽  
Endothelial Permeability ◽  
Endothelial Barrier ◽  
Endothelial Barrier Function ◽  
Barrier Dysfunction ◽  
Endothelial Barrier Dysfunction ◽  
Aryl Hydrocarbon ◽  
Underlying Mechanisms

Background: Diabetes leads to endothelial barrier dysfunction and altered endothelial permeability, which results in increased cardiovascular risk. ARNT, also known as HIF-1β, a transcription factor that functions as a master regulator of glucose homeostasis, has been implicated in diabetes. Endothelial-specific ARNT deletion (ArntΔEC) in mice is embryonically lethal, with hemorrhage occurring in the heart during the embryonic stage. However, the particular role of endothelial ARNT(ecARNT) in diabetes is largely unknown. We have found a significant decrease in ARNT expression in both diabetic rodent endothelial cells and diabetic human hearts. We hypothesize that a loss of ecARNT mediates endothelial barrier dysfunction during diabetes. Methods and Results: We generated inducible endothelial specific ARNT knockout mice (ecARNT-/-) by crossing mice with loxP sequences flanking exon 6 of ARNT with Cre ERT2 mice under the VE-cadherin promoter. A 90% deletion of ecARNT was achieved following two weeks of oral tamoxifen administration. ecARNT-/- mice exhibit severe blood vessel leakage, which is restricted to the heart, suggesting a distinct function for ecARNT in different tissues. Cardiomyopathy is evident 6 months after ARNT deletion. In vitro , trans-endothelial electrical resistance (TER) and transwell assays have confirmed endothelial barrier disruption in cardiac microvascular endothelial cells (CMEC) isolated from both ecARNT-/- hearts and diabetic (DB/DB) mouse hearts. To determine the underlying mechanisms by which ARNT may regulate endothelial barrier function, we performed DNA sequencing on CMEC isolated from control, ecARNT-/-, and DB/DB mice. Data suggest a significant increase in TNFa signaling, including ELAM-1 and ICAM-1 in CMEC isolated from ecARNT-/- CMEC and diabetic CMEC. Moreover, use of anti-TNFa antibody rescues endothelial barrier dysfunction in CMEC isolated from ecARNT-/- mice. Taken together, these results suggest that a reduction in ecARNT during diabetes may mediate endothelial barrier dysfunction through a TNFa signaling pathway. Conclusion: ecARNT is a critical mediator of endothelial barrier function and could potentially serve as a therapeutic target for diabetic cardiovascular diseases.

scholarly journals Impact of Atherosclerosis- and Diabetes-Related Dicarbonyls on Vascular Endothelial Permeability: A Comparative Assessment

2017 ◽  
Vol 2017 ◽  
pp. 1-10 ◽  
Author(s):  
Mikhail V. Samsonov ◽  
Asker Y. Khapchaev ◽  
Alexander V. Vorotnikov ◽  
Tatyana N. Vlasik ◽  
Elena V. Yanushevskaya ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Fluorescent Microscopy ◽  
Endothelial Permeability ◽  
Time Lapse ◽  
Protein Band ◽  
Endothelial Barrier ◽  
Vascular Endothelial ◽  
Barrier Dysfunction ◽  
Cytoskeletal Organization ◽  
Diabetes Patients

Background. Malondialdehyde (MDA), glyoxal (GO), and methylglyoxal (MGO) levels increase in atherosclerosis and diabetes patients. Recent reports demonstrate that GO and MGO cause vascular endothelial barrier dysfunction whereas no evidence is available for MDA. Methods. To compare the effects of MDA, GO, or MGO on endothelial permeability, we used human EA.hy926 endothelial cells as a standard model. To study cortical cytoplasm motility and cytoskeletal organization in endothelial cells, we utilized time-lapse microscopy and fluorescent microscopy. To compare dicarbonyl-modified protein band profiles in these cells, we applied Western blotting with antibodies against MDA- or MGO-labelled proteins. Results. MDA (150–250 μM) irreversibly suppressed the endothelial cell barrier, reduced lamellipodial activity, and prevented intercellular contact formation. The motile deficiency of MDA-challenged cells was accompanied by alterations in microtubule and microfilament organization. These detrimental effects were not observed after GO or MGO (250 μM) administration regardless of confirmed modification of cellular proteins by MGO. Conclusions. Our comparative study demonstrates that MDA is more damaging to the endothelial barrier than GO or MGO. Considering that MDA endogenous levels exceed those of GO or MGO and tend to increase further during lipoperoxidation, it appears important to reduce oxidative stress and, in particular, MDA levels in order to prevent sustained vascular hyperpermeability in atherosclerosis and diabetes patients.

scholarly journals Tongxinluo Boosts the Effect of Atorvastatin in Treatment of Atherosclerosis With COPD by Maintaining the Pulmonary Microvascular Barrier

2021 ◽  
Author(s):  
Xiangnan Kuang ◽  
Yafen Wang ◽  
Liping Chang ◽  
Yujie Yin ◽  
Zhen Li ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Systemic Inflammation ◽  
Barrier Function ◽  
High Fat Diet ◽  
Lung Inflammation ◽  
Endothelial Damage ◽  
Endothelial Barrier ◽  
High Fat ◽  
Barrier Dysfunction ◽  
Microvascular Endothelial

Abstract Background: Atherosclerosis (AS) is a common comorbidity of chronic obstructive pulmonary disease (COPD) and the main cause of death in patients with COPD. Systemic inflammation is a significant mechanism of COPD with AS. The dysfunction of pulmonary microvascular barrier is involved in the formation of chronic inflammation in COPD, and its functional disruption will induce systemic inflammation. Atorvastatin (Ato) is a common medicine for the treatment of AS; however, the effect is not ideal for COPD combined with AS. Tongxinluo (TXL) improves the function of vascular endothelial cells. This study aims to prove that the impairment of pulmonary microvascular barrier function participated in the process of COPD aggravating AS and investigated whether TXL enhances the therapeutic effect of Ato on COPD with AS by protecting the pulmonary microvascular endothelial barrier function. Methods: In vivo, the COPD with AS model of ApoE-/- mice was established by cigarette smoke combined with a high-fat diet. The animals were administered TXL (1.5 g/kg/day), Ato (10 mg/kg/day), and TXL+Ato once a day for 20 weeks. Then, lung function, lung microvascular permeability, lung inflammation, systemic inflammation, serum lipid level, atheromatous plaque formation, and the biomarker of endothelial damage were measured. In vitro, human pulmonary microvascular endothelial cells (HPMECs) were pretreated with TXL for 6 h and incubated with cigarette smoke extract (CSE) for 24 h to establish a model of CSE-induced pulmonary microvascular barrier dysfunction. The permeability of the endothelial monolayer, inflammatory cytokines, endothelial damage biomarkers, tight junction proteins were determined. Results: Cigarette smoking significantly exacerbated, which induced by high fat diet, the pulmonary function decline, pulmonary microvascular endothelial barrier dysfunction, pulmonary and systemic inflammation, and atherosclerotic plaque. These changes were reversed by TXL-Ato combination; the combination therapeutic effect was better than that of Ato alone. In addition, TXL protected the function of HPMEC barrier and inhibited the inflammation in CSE-induced HPMECs.Conclusions: COPD aggravates AS, the mechanism may be the destruction of pulmonary microvascular barrier function and thus lung inflammation triggers systemic inflammation. In treating COPD with AS, TXL enhances the anti-AS effect of Ato protecting the pulmonary microvascular barrier.

scholarly journals Roles of VE-Cadherin in Hypoxia Induced Injury of Pulmonary Microvascular Endothelial Barrier

2021 ◽  
Vol 24 (4) ◽  
pp. E764-E768
Author(s):  
Lizhe Zhong ◽  
Xiurong Gao ◽  
Yongli Chen ◽  
Zhaoxiang Yu ◽  
Shuo Jin ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Acute Lung Injury ◽  
Endothelial Barrier ◽  
Microvascular Endothelial Cells ◽  
Barrier Dysfunction ◽  
Barrier Methods ◽  
Endothelial Barrier Dysfunction ◽  
Microvascular Endothelial ◽  
Induced Apoptosis

Background: Hypoxia induced injury of pulmonary microvascular endothelial barrier is closely related to the pathogenesis of acute lung injury after lung transplantation. VE-cadherin is an important structural molecule for pulmonary microvascular endothelial barrier. In this study, we aim to investigate the roles of VE-cadherin in hypoxia induced injury of pulmonary microvascular endothelial barrier. Methods: Rat model of hypoxia and cultured pulmonary microvascular endothelial cells (PMVECs) were utilized. Determination of PMVECs apoptosis, skeleton combination was conducted to verify the effects of hypoxia on injury of pulmonary microvascular endothelial barrier. In addition, VE-cadherin expression was modulated by administration of siRNA in order to investigate the roles of VE-cadherin in hypoxia induced PMVECs apoptosis and skeleton recombination. Results: Our data indicated that expression of VE-cadherin was down-regulated in hypoxia-exposed PMVECs. Whereas, in the cells treated using siRNA, down-regulation of VE-cadherin did not trigger PMVECs apoptosis, but it increased the sensitivity of PMVECs to the hypoxia induced apoptosis. In cases of hypoxia, the expression of VE-cadherin was significantly down-regulated, together with endothelial skeleton recombination and increase of permeability, which then triggered endothelial barrier dysfunction. Conclusions: These data verify that VE-cadherin expression played an important role in hypoxia induced PMVECs apoptosis and cellular skeletal recombination.

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