Loss of endothelial sulfatase-1 after experimental sepsis attenuates subsequent pulmonary inflammatory responses

Sepsis patients are at increased risk for hospital-acquired pulmonary infections, potentially due to postseptic immunosuppression known as the compensatory anti-inflammatory response syndrome (CARS). CARS has been attributed to leukocyte dysfunction, with an unclear role for endothelial cells. The pulmonary circulation is lined by an endothelial glycocalyx, a heparan sulfate-rich layer essential to pulmonary homeostasis. Heparan sulfate degradation occurs early in sepsis, leading to lung injury. Endothelial synthesis of new heparan sulfates subsequently allows for glycocalyx reconstitution and endothelial recovery. We hypothesized that remodeling of the reconstituted endothelial glycocalyx, mediated by alterations in the endothelial machinery responsible for heparan sulfate synthesis, contributes to CARS. Seventy-two hours after experimental sepsis, coincident with glycocalyx reconstitution, mice demonstrated impaired neutrophil and protein influx in response to intratracheal lipopolysaccharide (LPS). The postseptic reconstituted glycocalyx was structurally remodeled, with enrichment of heparan sulfate disaccharides sulfated at the 6- O position of glucosamine. Increased 6- O-sulfation coincided with loss of endothelial sulfatase-1 (Sulf-1), an enzyme that specifically removes 6- O-sulfates from heparan sulfate. Intravenous administration of Sulf-1 to postseptic mice restored the pulmonary response to LPS, suggesting that loss of Sulf-1 was necessary for postseptic suppression of pulmonary inflammation. Endothelial-specific knockout mice demonstrated that loss of Sulf-1 was not sufficient to induce immunosuppression in non-septic mice. Knockdown of Sulf-1 in human pulmonary microvascular endothelial cells resulted in downregulation of the adhesion molecule ICAM-1. Taken together, our study indicates that loss of endothelial Sulf-1 is necessary for postseptic suppression of pulmonary inflammation, representing a novel endothelial contributor to CARS.

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Suppressive Effects of Sulforaphane on TGFBIp-mediated Sepsis

Natural Product Communications ◽

10.1177/1934578x1701201026 ◽

2017 ◽

Vol 12 (10) ◽

pp. 1934578X1701201 ◽

Cited By ~ 1

Author(s):

In-Chul Lee ◽

Jong-Sup Bae

Keyword(s):

Endothelial Cells ◽

Matrix Protein ◽

Transforming Growth Factor ◽

Inflammatory Responses ◽

Inflammatory Diseases ◽

Umbilical Vein ◽

Cecal Ligation ◽

Transforming Growth Factor Β ◽

Extracellular Matrix Protein ◽

Experimental Sepsis

Sulforaphane (SFN) is produced when the enzyme myrosinase transforms glucoraphanin upon damage to the plant such as from chewing and effective in preventing carcinogenesis, diabetes, and inflammatory responses. Transforming growth factor β-induced protein (TGFBIp) is an extracellular matrix protein whose expression in several cell types is greatly increased by TGF-β. TGFBIp is released by human umbilical vein endothelial cells (HUVECs) and functions as a mediator of experimental sepsis. We hypothesized that SFN could reduce TGFBIp-mediated severe inflammatory responses in human endothelial cells and mice. Here, we investigated the anti-septic effects and underlying mechanisms of SFN against TGFBIp-mediated septic responses. SFN effectively inhibited lipopolysaccharide-induced release of TGFBIp and suppressed TGFBIp-mediated septic responses. In addition, SFN suppressed cecal ligation and puncture (CLP)-induced sepsis lethality and pulmonary injury. In conclusion, SFN suppressed TGFBIp-mediated and CLP-induced septic responses. Therefore, SFN could be a potential therapeutic agent for treatment of various severe vascular inflammatory diseases via inhibition of the TGFBIp signaling pathway.

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Pulmonary delivery of the broad-spectrum matrix metalloproteinase inhibitor marimastat diminishes multiwalled carbon nanotube-induced circulating bioactivity without reducing pulmonary inflammation

Particle and Fibre Toxicology ◽

10.1186/s12989-021-00427-w ◽

2021 ◽

Vol 18 (1) ◽

Author(s):

Tamara L. Young ◽

Ekaterina Mostovenko ◽

Jesse L. Denson ◽

Jessica G. Begay ◽

Selita N. Lucas ◽

...

Keyword(s):

Endothelial Cells ◽

Matrix Metalloproteinase ◽

Broad Spectrum ◽

Inflammatory Responses ◽

Pulmonary Inflammation ◽

Significant Risk ◽

Multiwalled Carbon ◽

Endothelial Cell Function ◽

Serum Peptide ◽

Oropharyngeal Aspiration

Abstract Background Multiwalled carbon nanotubes (MWCNT) are an increasingly utilized engineered nanomaterial that pose the potential for significant risk of exposure-related health outcomes. The mechanism(s) underlying MWCNT-induced toxicity to extrapulmonary sites are still being defined. MWCNT-induced serum-borne bioactivity appears to dysregulate systemic endothelial cell function. The serum compositional changes after MWCNT exposure have been identified as a surge of fragmented endogenous peptides, likely derived from matrix metalloproteinase (MMP) activity. In the present study, we utilize a broad-spectrum MMP inhibitor, Marimastat, along with a previously described oropharyngeal aspiration model of MWCNT administration to investigate the role of MMPs in MWCNT-derived serum peptide generation and endothelial bioactivity. Results C57BL/6 mice were treated with Marimastat or vehicle by oropharyngeal aspiration 1 h prior to MWCNT treatment. Pulmonary neutrophil infiltration and total bronchoalveolar lavage fluid protein increased independent of MMP blockade. The lung cytokine profile similarly increased following MWCNT exposure for major inflammatory markers (IL-1β, IL-6, and TNF-α), with minimal impact from MMP inhibition. However, serum peptidomic analysis revealed differential peptide compositional profiles, with MMP blockade abrogating MWCNT-derived serum peptide fragments. The serum, in turn, exhibited differential potency in terms of inflammatory bioactivity when incubated with primary murine cerebrovascular endothelial cells. Serum from MWCNT-treated mice led to inflammatory responses in endothelial cells that were significantly blunted with serum from Marimastat-treated mice. Conclusions Thus, MWCNT exposure induced pulmonary inflammation that was largely independent of MMP activity but generated circulating bioactive peptides through predominantly MMP-dependent pathways. This MWCNT-induced lung-derived bioactivity caused pathological consequences of endothelial inflammation and barrier disruption.

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Inverse Regulation of Confluence-Dependent ADAMTS13 and von Willebrand Factor Expression in Human Endothelial Cells

Thrombosis and Haemostasis ◽

10.1055/s-0041-1733800 ◽

2021 ◽

Author(s):

Miruna Popa ◽

Markus Hecker ◽

Andreas H. Wagner

Keyword(s):

Endothelial Cells ◽

Shear Stress ◽

Von Willebrand Factor ◽

Interleukin 10 ◽

Endothelial Glycocalyx ◽

Human Endothelial Cells ◽

Proliferating Cells ◽

Increased Risk ◽

Von Willebrand ◽

Willebrand Factor

AbstractADAMTS13 (a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13) is a zinc-containing metalloprotease also known as von Willebrand factor (vWF)-cleaving protease. Low ADAMTS13 plasma levels are associated with an increased risk of arterial thrombosis, including myocardial infarction and cerebrovascular disease. The expression and regulation of this metalloprotease in human endothelial cells have not been systematically investigated. In this study, we demonstrate that ADAMTS13 expression is inhibited by proinflammatory cytokines tumor necrosis factor-α and interferon-γ as well as by CD40 ligand, which was hitherto unknown. Factors protecting against atherosclerosis such as exposure to continuous unidirectional shear stress, interleukin-10, or different HMG-CoA reductase inhibitors like, e.g., simvastatin, atorvastatin, or rosuvastatin, did not influence ADAMTS13 expression. Unidirectional periodic orbital shear stress, mimicking oscillatory flow conditions found at atherosclerosis-prone arterial bifurcations, had also no effect. In contrast, a reciprocal correlation between ADAMTS13 and vWF expression in endothelial cells depending on the differentiation state was noted. ADAMTS13 abundance significantly rose on both the mRNA and intracellular protein level and also tethered to the endothelial glycocalyx with the degree of confluency while vWF protein levels were highest in proliferating cells but significantly decreased upon reaching confluence. This finding could explain the anti-inflammatory and antithrombotic phenotype of dormant endothelial cells mediated by contact inhibition.

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Matrix Stiffness Affects Glycocalyx Expression in Cultured Endothelial Cells

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.731666 ◽

2021 ◽

Vol 9 ◽

Author(s):

Marwa Mahmoud ◽

Limary Cancel ◽

John M. Tarbell

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

Heparan Sulfate ◽

Cell Types ◽

Endothelial Glycocalyx ◽

Specific Cell ◽

Matrix Stiffness ◽

Endothelial Cell Function ◽

Mouse Brain Endothelial Cells ◽

Aortic Endothelial

Rationale: The endothelial cell glycocalyx (GCX) is a mechanosensor that plays a key role in protecting against vascular diseases. We have previously shown that age/disease mediated matrix stiffness inhibits the glycocalyx glycosaminoglycan heparan sulfate and its core protein Glypican 1 in human umbilical vein endothelial cells, rat fat pad endothelial cells and in a mouse model of age-mediated stiffness. Glypican 1 inhibition resulted in enhanced endothelial cell dysfunction. Endothelial cell culture typically occurs on stiff matrices such as plastic or glass. For the study of the endothelial GCX specifically it is important to culture cells on soft matrices to preserve GCX expression. To test the generality of this statement, we hypothesized that stiff matrices inhibit GCX expression and consequently endothelial cell function in additional cell types: bovine aortic endothelial cells, mouse aortic endothelial cell and mouse brain endothelial cells.Methods and Results: All cell types cultured on glass showed reduced GCX heparan sulfate expression compared to cells cultured on either soft polyacrylamide (PA) gels of a substrate stiffness of 2.5 kPa (mimicking the stiffness of young, healthy arteries) or on either stiff gels 10 kPa (mimicking the stiffness of old, diseased arteries). Specific cell types showed reduced expression of GCX protein Glypican 1 (4 of 5 cell types) and hyaluronic acid (2 of 5 cell types) on glass vs soft gels.Conclusion: Matrix stiffness affects GCX expression in endothelial cells. Therefore, the study of the endothelial glycocalyx on stiff matrices (glass/plastic) is not recommended for specific cell types.

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Heparan sulfates mediate pressure-induced increase in lung endothelial hydraulic conductivity via nitric oxide/reactive oxygen species

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00376.2006 ◽

2007 ◽

Vol 292 (6) ◽

pp. L1452-L1458 ◽

Cited By ~ 41

Author(s):

Randal O. Dull ◽

Ian Mecham ◽

Scott McJames

Keyword(s):

Nitric Oxide ◽

Reactive Oxygen Species ◽

Endothelial Cells ◽

Hydraulic Conductivity ◽

Reactive Oxygen ◽

Endothelial Glycocalyx ◽

Oxygen Species ◽

Barrier Dysfunction ◽

Heparan Sulfates ◽

Intracellular Fluorescence

We investigated the nonlinear dynamics of the pressure vs. hydraulic conductivity (Lp) relationship in lung microvascular endothelial cells and demonstrate that heparan sulfates, an important component of the endothelial glycocalyx, participate in pressure-sensitive mechanotransduction that results in barrier dysfunction. The pressure vs. Lp relationship was complex, possessing both time- and pressure-dependent components. Pretreatment of lung capillary endothelial cells with heparanase III completely abolished the pressure-induced increase in Lp. This extends our ( 7 ) previous observation regarding heparan sulfates as mechanotransducers for shear stress. Inhibition of nitric oxide (NO) synthase with l-NAME ( NG-nitro-l-arginine methyl ester HCl) and intracellular scavenging of reactive oxygen species (ROS) by TBAP [tetrakis-(4-benzoic acid) porphorin] significantly attenuated the pressure-induced Lp response. Intracellular NO/ROS were visualized using the fluorescent dye, 2′7′-dichlorofluorescein diacetate (DCFA), and cells demonstrated a pressure-induced increase in intracellular fluorescence. Heparanase pretreatment significantly reduced the pressure-induced increase in intracellular fluorescence, suggesting that cell-surface heparan sulfates directly participate in mechanotransduction that results in NO/ROS production and increased permeability. This is the first report to demonstrate a role for heparan sulfates in pressure-mediated mechanotransduction and barrier regulation. These observations may have important clinical implications during conditions where pulmonary microvascular pressure is elevated.

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Obesity and lung inflammation

Journal of Applied Physiology ◽

10.1152/japplphysiol.00781.2009 ◽

2010 ◽

Vol 108 (3) ◽

pp. 722-728 ◽

Cited By ~ 99

Author(s):

Peter Mancuso

Keyword(s):

Adipose Tissue ◽

Lung Disease ◽

Inflammatory Responses ◽

Acute Phase Proteins ◽

Pulmonary Inflammation ◽

Therapeutic Interventions ◽

Low Grade ◽

Tissue Mass ◽

Proinflammatory Mediators ◽

Increased Risk

The prevalence of obesity has increased dramatically worldwide, predisposing individuals to an increased risk of morbidity and mortality due to cardiovascular disease and type 2 diabetes. Less recognized is the fact that obesity may play a significant role in the pathogenesis of pulmonary diseases through mechanisms that may involve proinflammatory mediators produced in adipose tissue that contribute to a low-grade state of systemic inflammation. In animal models, inflammatory responses in the lung have been shown to influence the production of the adipocytokines, leptin and adiponectin, cytokines, acute phase proteins, and other mediators produced by adipose tissue that may participate in immune responses of the lung. An increased adipose tissue mass may also influence susceptibility to pulmonary infections, enhance pulmonary inflammation associated with environmental exposures, and exacerbate airway obstruction in preexisting lung disease. An increased understanding of the mechanisms by which obesity influences pulmonary inflammation may facilitate the development of novel therapeutic interventions for the treatment of lung disease.

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Unraveling the Complex Relationship Triad between Lipids, Obesity, and Inflammation

Mediators of Inflammation ◽

10.1155/2014/502749 ◽

2014 ◽

Vol 2014 ◽

pp. 1-16 ◽

Cited By ~ 32

Author(s):

Shahida A. Khan ◽

Ashraf Ali ◽

Sarah A. Khan ◽

Solafa A. Zahran ◽

Ghazi Damanhouri ◽

...

Keyword(s):

Inflammatory Responses ◽

Pulmonary Inflammation ◽

Lipid Mediators ◽

Peroxisome Proliferator ◽

Omega 3 ◽

Mortality And Morbidity ◽

Increased Risk ◽

Omega 6 ◽

Anti Inflammatory ◽

Peroxisome Proliferator Activated Receptors

Obesity today stands at the intersection between inflammation and metabolic disorders causing an aberration of immune activity, and resulting in increased risk for diabetes, atherosclerosis, fatty liver, and pulmonary inflammation to name a few. Increases in mortality and morbidity in obesity related inflammation have initiated studies to explore different lipid mediated molecular pathways of attempting resolution that uncover newer therapeutic opportunities of anti-inflammatory components. Majorly the thromboxanes, prostaglandins, leukotrienes, lipoxins, and so forth form the group of lipid mediators influencing inflammation. Of special mention are the omega-6 and omega-3 fatty acids that regulate inflammatory mediators of interest in hepatocytes and adipocytes via the cyclooxygenase and lipoxygenase pathways. They also exhibit profound effects on eicosanoid production. The inflammatory cyclooxygenase pathway arising from arachidonic acid is a critical step in the progression of inflammatory responses. New oxygenated products of omega-3 metabolism, namely, resolvins and protectins, behave as endogenous mediators exhibiting powerful anti-inflammatory and immune-regulatory actions via the peroxisome proliferator-activated receptors (PPARs) and G protein coupled receptors (GPCRs). In this review we attempt to discuss the complex pathways and links between obesity and inflammation particularly in relation to different lipid mediators.

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Ischemic stroke disrupts the endothelial glycocalyx through activation of proHPSE via acrolein exposure

Journal of Biological Chemistry ◽

10.1074/jbc.ra120.015105 ◽

2020 ◽

Vol 295 (52) ◽

pp. 18614-18624

Author(s):

Kenta Ko ◽

Takehiro Suzuki ◽

Ryota Ishikawa ◽

Natsuko Hattori ◽

Risako Ito ◽

...

Keyword(s):

Endothelial Cells ◽

Ischemic Stroke ◽

Heparan Sulfate ◽

Infarct Volume ◽

Hek293 Cells ◽

Endothelial Glycocalyx ◽

Stroke Severity ◽

Amino Acid Residues ◽

Barrier Dysfunction ◽

Promising Target

Infiltration of peripheral immune cells after blood-brain barrier dysfunction causes severe inflammation after a stroke. Although the endothelial glycocalyx, a network of membrane-bound glycoproteins and proteoglycans that covers the lumen of endothelial cells, functions as a barrier to circulating cells, the relationship between stroke severity and glycocalyx dysfunction remains unclear. In this study, glycosaminoglycans, a component of the endothelial glycocalyx, were studied in the context of ischemic stroke using a photochemically induced thrombosis mouse model. Decreased levels of heparan sulfate and chondroitin sulfate and increased activity of hyaluronidase 1 and heparanase (HPSE) were observed in ischemic brain tissues. HPSE expression in cerebral vessels increased after stroke onset and infarct volume greatly decreased after co-administration of N-acetylcysteine + glycosaminoglycan oligosaccharides as compared with N-acetylcysteine administration alone. These results suggest that the endothelial glycocalyx was injured after the onset of stroke. Interestingly, scission activity of proHPSE produced by immortalized endothelial cells and HEK293 cells transfected with hHPSE1 cDNA were activated by acrolein (ACR) exposure. We identified the ACR-modified amino acid residues of proHPSE using nano LC–MS/MS, suggesting that ACR modification of Lys139 (6-kDa linker), Lys107, and Lys161, located in the immediate vicinity of the 6-kDa linker, at least in part is attributed to the activation of proHPSE. Because proHPSE, but not HPSE, localizes outside cells by binding with heparan sulfate proteoglycans, ACR-modified proHPSE represents a promising target to protect the endothelial glycocalyx.

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On, Around, and Through: Neutrophil-Endothelial Interactions in Innate Immunity

Physiology ◽

10.1152/physiol.00011.2011 ◽

2011 ◽

Vol 26 (5) ◽

pp. 334-347 ◽

Cited By ~ 41

Author(s):

Eric P. Schmidt ◽

Warren L. Lee ◽

Rachel L. Zemans ◽

Cory Yamashita ◽

Gregory P. Downey

Keyword(s):

Endothelial Cells ◽

Molecular Mechanisms ◽

Inflammatory Responses ◽

Tissue Injury ◽

Endothelial Glycocalyx ◽

Neutrophil Adhesion ◽

Complex Field ◽

Interested Reader ◽

And Migration

This manuscript will review our current understanding of neutrophilic polymorphonuclear leukocyte (neutrophil) interactions with the endothelium during immune and inflammatory responses, focusing on the molecular mechanisms regulating neutrophil adhesion to and migration through the endothelium in response to infection or tissue injury. This is a complex and dynamic area of research and one that has been the topic of several recent comprehensive reviews to which the interested reader is referred ( 64 , 118 , 131 ). By design, this review will begin with a brief review of some basic aspects of neutrophil biology and endothelial adhesion to provide a foundation. The remainder of the review will focus on selected areas of this complex field, specifically the role of the endothelial glycocalyx in regulating neutrophil adhesion and the mechanisms and consequences of migration of neutrophils between (paracellular) and through (transcellular) endothelial cells during egress from the vasculature.

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Hemodynamic shear stress characteristic of atherosclerosis-resistant regions promotes glycocalyx formation in cultured endothelial cells

AJP Cell Physiology ◽

10.1152/ajpcell.00187.2012 ◽

2013 ◽

Vol 304 (2) ◽

pp. C137-C146 ◽

Cited By ~ 75

Author(s):

Andrew Koo ◽

C. Forbes Dewey ◽

Guillermo García-Cardeña

Keyword(s):

Endothelial Cells ◽

Shear Stress ◽

Heparan Sulfate ◽

Vascular Endothelial Cells ◽

Surface Expression ◽

Endothelial Glycocalyx ◽

Cell Surface Expression ◽

Apical Surface ◽

Laminar Shear Stress ◽

Critical Components

The endothelial glycocalyx, a glycosaminoglycan layer located on the apical surface of vascular endothelial cells, has been shown to be important for several endothelial functions. Previous studies have documented that the glycocalyx is highly abundant in the mouse common carotid region, where the endothelium is exposed to laminar shear stress, and it is resistant to atherosclerosis. In contrast, the glycocalyx is scarce or absent in the mouse internal carotid sinus region, an area exposed to nonlaminar shear stress and highly susceptible to atherosclerosis. On the basis of these observations, we hypothesized that the expression of components of the endothelial glycocalyx is differentially regulated by distinct hemodynamic environments. To test this hypothesis, human endothelial cells were exposed to shear stress waveforms characteristic of atherosclerosis-resistant or atherosclerosis-susceptible regions of the human carotid, and the expression of several components of the glycocalyx was assessed. These experiments revealed that expression of several components of the endothelial glycocalyx is differentially regulated by distinct shear stress waveforms. Interestingly, we found that heparan sulfate expression is increased and evenly distributed on the apical surface of endothelial cells exposed to the atheroprotective waveform and is irregularly present in cells exposed to the atheroprone waveform. Furthermore, expression of a heparan sulfate proteoglycan, syndecan-1, is also differentially regulated by the two waveforms, and its suppression mutes the atheroprotective flow-induced cell surface expression of heparan sulfate. Collectively, these data link distinct hemodynamic environments to the differential expression of critical components of the endothelial glycocalyx.

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