German Cockroach Extract Induces Matrix Metalloproteinase-1 Expression, Leading to Tight Junction Disruption in Human Airway Epithelial Cells.
10.3349/ymj.2018.59.10.1222
- Author:
Kyung Eun LEE
1
;
Hye Mi JEE
;
Jung Yeon HONG
;
Mi Na KIM
;
Mi Seon OH
;
Yun Seon KIM
;
Kyung Won KIM
;
Kyu Earn KIM
;
Myung Hyun SOHN
Author Information
1. Department of Pediatrics, Institute of Allergy, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea. mhsohn@yuhs.ac
- Publication Type:Original Article
- Keywords:
German cockroach;
asthma;
matrix metalloproteinase;
airway epithelial cell;
tight junction
- MeSH:
Airway Obstruction;
Asthma;
Blattellidae*;
Blotting, Western;
Cockroaches;
Collagenases;
DNA;
Electric Impedance;
Electrophoretic Mobility Shift Assay;
Enzyme-Linked Immunosorbent Assay;
Epithelial Cells*;
Epithelium;
Extracellular Matrix;
Fluorescent Antibody Technique;
Humans*;
Matrix Metalloproteinase 1*;
Matrix Metalloproteinases;
Phosphorylation;
Phosphotransferases;
Protein Kinases;
Real-Time Polymerase Chain Reaction;
RNA, Messenger;
RNA, Small Interfering;
Tight Junction Proteins;
Tight Junctions*;
Transcription Factors;
Transfection
- From:Yonsei Medical Journal
2018;59(10):1222-1231
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: Cockroach 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 METHODS: mRNA 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. RESULTS: GCE 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. CONCLUSION: GCE 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.
