German Cockroach Extract Induces Matrix Metalloproteinase-1 Expression, Leading to Tight Junction Disruption in Human Airway Epithelial Cells

Abstract

PurposeCockroach exposure is a pivotal cause of asthma. Tight junctions are intercellular structures required for maintenance of the barrier function of the airway epithelium, which is impaired in this disease. Matrix metalloproteinases (MMPs) digest extracellular matrix components and are involved in asthma pathogenesis: MMP1 is a collagenase with a direct influence on airway obstruction in asthmatics. This study aimed to investigate the mechanism by which German cockroach extract (GCE) induces MMP1 expression and whether MMP1 release alters cellular tight junctions in human airway epithelial cells (NCI-H292).Materials and MethodsmRNA and protein levels were determined using real-time PCR and ELISA. Tight junction proteins were detected using immunofluorescence staining. Epithelial barrier function was measured by transepithelial electrical resistance (TEER). The binding of a transcription factor to DNA molecules was determined by electrophoretic mobility shift assay, while the levels of tight junction proteins and phosphorylation were determined using Western blotting.ResultsGCE was shown to increase MMP1 expression, TEER, and tight junction degradation. Both an inhibitor and small interfering RNA (siRNA) of MMP1 significantly decreased GCE-induced tight junction disruption. Furthermore, transient transfection with ETS1 and SP1 siRNA, and anti-TLR2 antibody pretreatment prevented MMP1 expression and tight junction degradation. An extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase (MAPK) inhibitor also blocked MMP1 release, ETS1/SP1 DNA binding, and tight junction alteration.ConclusionGCE treatment increases MMP1 expression, leading to tight junction disruption, which is transcriptionally regulated and influenced by the ERK/MAPK pathway in airway epithelial cells. These findings may contribute to developing novel therapeutic strategies for airway diseases.

Modeling CADASIL vascular pathologies with patient-derived induced pluripotent stem cells

“…For imaging and analysis of cytoskeletal structures, the statement of the normal cytoskeletal structures were based on the morphology of the WTs in the representative picture. The normal cytoskeletal structures are as follows: the microfilaments should without obvious robust bundles or nodular structures (Domenga et al, 2004 ; Tikka et al, 2012 ); the intermediate filaments and microtubules should be in the form of a filament-like structure distributed in a network without significant aggregation (Fogl et al, 2016 ; Fuertes-Alvarez et al, 2018 ; Tu et al, 2018 ); the adhesion junction protein vinculin should be in a punctiform structure and co-localizes with the microfilaments near the cell membrane (Tikka et al, 2012 ); the tight junction proteins are membrane-localized proteins that resemble the pattern of other VEC surface markers such as CD31 and CD144 (Lee et al, 2018 ; Zhang et al, 2018b ).…”

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“…Olea europaea , Dactylis glomerata , Cupressus sempervirens , and Pinus sylvestris allergens also induce variable E-cadherin, OCLN, and claudin-1 disruption in Calu-3 cells ALI-cultures ( Vinhas et al, 2011 ). Cockroach allergen extracts also show complex proteolytic activities, notably through Per a 10 (serine protease) and Bla g 2 (aspartic protease) ( Pomes et al, 2002 ; Sudha et al, 2008 ), but their action on the epithelial barrier stricto sensu is still unclear, although decreased TER in cockroach extract antigen-exposed BEAS-2 and ATCC cells (monolayer cell culture) has been reported ( Antony et al, 2002 ; Lee et al, 2018 ). Besides its hypothetical effects on barrier permeability, Per a 10 induces IL-6, IL-8/CXCL-8, and GM-CSF in A549 cells ( Kale and Arora, 2015 ).…”