Quantification of adherens junction disruption and contiguous paracellular protein leak in human lung endothelial cells under septic conditions.
Microcirculation
; 26(3): e12528, 2019 04.
Article
em En
| MEDLINE
| ID: mdl-30636088
ABSTRACT
OBJECTIVE:
Sepsis is associated with dysfunction of MVEC resulting in organ edema and inflammation. VE-cadherin, a component of MVEC adherens junctions, may be disrupted in sepsis. However, the direct connection between individual MVEC VE-cadherin disruption and increased paracellular permeability is uncertain.METHODS:
Human pulmonary MVEC were cultured on a biotin matrix and treated with cytomix, as a model of sepsis, vs PBS. MVEC permeability was assessed by trans-MVEC monolayer leak of Oregon green 488-conjugated avidin, which bound subcellular biotin to localize sites of paracellular leak. Leak was correlated with individual cell-specific MVEC surface VE-cadherin continuity by fluorescence microscopy.RESULTS:
Cytomix treatment reduced total MVEC VE-cadherin density, disrupted surface VE-cadherin continuity, was associated with intercellular gap formation, and enhanced paracellular avidin leak. Cytomix-induced MVEC paracellular avidin leak was strongly correlated temporally and was highly contiguous with focal MVEC surface VE-cadherin disruption. Total cellular VE-cadherin density was less strongly correlated with MVEC paracellular avidin leak and individual cell-specific focal surface VE-cadherin discontinuity.CONCLUSIONS:
These data support a mechanistic link between septic human lung MVEC VE-cadherin disruption and contiguous paracellular protein leak, and will permit more detailed assessment of individual cell-specific mechanisms of septic MVEC barrier dysfunction.Palavras-chave
Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Permeabilidade Capilar
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Antígenos CD
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Caderinas
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Sepse
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Junções Aderentes
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Células Endoteliais
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Pulmão
Limite:
Humans
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article