Pseudomonas aeruginosa elastase causes transient disruption of tight junctions and downregulation of PAR-2 in human nasal epithelial cells

Respiratory Research

Takashi Kojima¹  Tetsuo Himi²  Norimasa Sawada³  Takayuki Kohno¹  Ken-ichi Takano²  Satoshi Hirakawa⁴  Ryo Miyata¹  Takashi Keira³  Kazufumi Obata²  Kazuaki Nomura³ 

[1] Department of Cell Science, Research Institute for Frontier Medicine, Sapporo Medical University School of Medicine, South-1, West-17, Chuo-ku, Sapporo 060-8556, Japan;Departments of Otolaryngology, Sapporo Medical University School of Medicine, Sapporo 060-8556, Japan;Departments of Pathology, Sapporo Medical University School of Medicine, Sapporo 060-8556, Japan;Departments of Pediatrics, Sapporo Medical University School of Medicine, Sapporo 060-8556, Japan

关键词: PAR-2;    Signal transduction;    Human nasal epithelial cells;    Barrier function;    Tight junctions;    Pseudomonas aeruginosa elastase;   

Others  :  790446
DOI  :  10.1186/1465-9921-15-21

 received in 2013-11-10, accepted in 2014-01-31,  发布年份 2014

【 摘 要 】

Background

Pseudomonas aeruginosa causes chronic respiratory disease, and the elastase enzyme that it produces increases the permeability of airway epithelial cells owing to the disruption of tight junctions. P. aeruginosa is also implicated in prolonged chronic rhinosinusitis. However, the effects of P. aeruginosa elastase (PE) against the barrier formed by human nasal epithelial cells (HNECs) remain unknown.

Methods

To investigate the mechanisms involved in the disruption of tight junctions by PE in HNECs, primary cultures of HNECs transfected with human telomerase reverse transcriptase (hTERT-HNECs) were used. The hTERT-HNECs were pretreated with inhibitors of various signal transduction pathways, PKC, MAPK, p38MAPK, PI3K, JNK, NF-κB, EGF receptor, proteasome, COX1 and COX2 before treatment with PE. Some cells were pretreated with siRNA and agonist of protease activated receptor-2 (PAR-2) before treatment with PE. Expression and structures of tight junctions were determined by Western blotting, real-time PCR, immunostaining and freeze-fracture. Transepithelial electrical resistance (TER) was examined as the epithelial barrier function.

Results

PE treatment transiently disrupted the epithelial barrier and downregulated the transmembrane proteins claudin-1 and -4, occludin, and tricellulin, but not the scaffold PDZ-expression proteins ZO-1 and -2 and adherens junction proteins E-cadherin and β-catenin. The transient downregulation of tight junction proteins was controlled via distinct signal transduction pathways such as the PKC, MAPK, PI3K, p38 MAPK, JNK, COX-1 and -2, and NF-κB pathways. Furthermore, treatment with PE transiently decreased PAR-2 expression, which also regulated the expression of the tight junction proteins. Treatment with a PAR-2 agonist prevented the downregulation of the tight junction proteins after PE treatment in HNECs.

Conclusions

PE transiently disrupts tight junctions in HNECs and downregulates PAR-2. The transient disruption of tight junctions by PE might occur repeatedly during chronic rhinosinusitis.

【 授权许可】

   
2014 Nomura et al.; licensee BioMed Central Ltd.

【 图 表 】

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