Nuclear factor erythroid 2-related factor 2 (Nrf2) regulates airway epithelial barrier integrity
Background: Inhaled corticosteroids enhance airway epithelial barrier integrity. However, the mechanism by which they accomplish this is unclear. Therefore, we investigated steroid-inducible genes and signaling pathways that were involved in enhancing airway epithelial barrier integrity. Methods: A...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2015-09-01
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| Series: | Allergology International |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1323893015001409 |
| _version_ | 1818542324215447552 |
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| author | Yoshitaka Shintani Shuichiro Maruoka Yasuhiro Gon Daisuke Koyama Akiko Yoshida Yutaka Kozu Kazumichi Kuroda Ikuko Takeshita Eriko Tsuboi Kaori Soda Shu Hashimoto |
| author_facet | Yoshitaka Shintani Shuichiro Maruoka Yasuhiro Gon Daisuke Koyama Akiko Yoshida Yutaka Kozu Kazumichi Kuroda Ikuko Takeshita Eriko Tsuboi Kaori Soda Shu Hashimoto |
| author_sort | Yoshitaka Shintani |
| collection | DOAJ |
| description | Background: Inhaled corticosteroids enhance airway epithelial barrier integrity. However, the mechanism by which they accomplish this is unclear. Therefore, we investigated steroid-inducible genes and signaling pathways that were involved in enhancing airway epithelial barrier integrity.
Methods: A human bronchial epithelial cell line (16HBE cells) was cultured with 10−6 M dexamethasone (DEX) for 3 days to enhance epithelial barrier integrity. After measuring transepithelial electrical resistance (TER) and paracellular permeability, we extracted total RNA from 16HBE cells and performed microarray and pathway analysis. After we identified candidate genes and a canonical pathway, we measured TER and immunostained for tight junction (TJ) and adherent junction (AJ) proteins in cells that had been transfected with specific small interfering RNAs (siRNAs) for these genes.
Results: We identified a nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated oxidative stress response pathway which was primarily involved in the steroid-induced enhancement of airway epithelial barrier integrity. Transfecting cells with Nrf2 specific siRNA reduced the steroid-induced enhancement of airway epithelial barrier integrity and the accumulation of TJ and AJ proteins at sites of cell–cell contact. Moreover, based on pathway analysis, aldehyde oxidase 1 (AOX1) was identified as a downstream enzyme of Nrf2. Transfecting cells with AOX1-specific siRNA also reduced the steroid-induced enhancement of airway epithelial barrier integrity.
Conclusions: Our results indicated that the Nrf2/AOX1 pathway was important for enhancing airway epithelial barrier integrity. Because the airway epithelium of asthmatics is susceptible to reduced barrier integrity, this pathway might be a new therapeutic target for asthma. |
| first_indexed | 2024-12-11T22:20:35Z |
| format | Article |
| id | doaj.art-4bf78a2e80f34bd9991ab2f83c541d32 |
| institution | Directory Open Access Journal |
| issn | 1323-8930 |
| language | English |
| last_indexed | 2024-12-11T22:20:35Z |
| publishDate | 2015-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Allergology International |
| spelling | doaj.art-4bf78a2e80f34bd9991ab2f83c541d322022-12-22T00:48:27ZengElsevierAllergology International1323-89302015-09-0164SS54S6310.1016/j.alit.2015.06.004Nuclear factor erythroid 2-related factor 2 (Nrf2) regulates airway epithelial barrier integrityYoshitaka Shintani0Shuichiro Maruoka1Yasuhiro Gon2Daisuke Koyama3Akiko Yoshida4Yutaka Kozu5Kazumichi Kuroda6Ikuko Takeshita7Eriko Tsuboi8Kaori Soda9Shu Hashimoto10Division of Respiratory Medicine, Department of Internal Medicine, Nihon University School of Medicine, Tokyo, JapanDivision of Respiratory Medicine, Department of Internal Medicine, Nihon University School of Medicine, Tokyo, JapanDivision of Respiratory Medicine, Department of Internal Medicine, Nihon University School of Medicine, Tokyo, JapanDivision of Respiratory Medicine, Department of Internal Medicine, Nihon University School of Medicine, Tokyo, JapanDivision of Respiratory Medicine, Department of Internal Medicine, Nihon University School of Medicine, Tokyo, JapanDivision of Respiratory Medicine, Department of Internal Medicine, Nihon University School of Medicine, Tokyo, JapanDivision of Microbiology, Department of Pathology and Microbiology, Nihon University School of Medicine, Tokyo, JapanDivision of Respiratory Medicine, Department of Internal Medicine, Nihon University School of Medicine, Tokyo, JapanDivision of Respiratory Medicine, Department of Internal Medicine, Nihon University School of Medicine, Tokyo, JapanDivision of Respiratory Medicine, Department of Internal Medicine, Nihon University School of Medicine, Tokyo, JapanDivision of Respiratory Medicine, Department of Internal Medicine, Nihon University School of Medicine, Tokyo, JapanBackground: Inhaled corticosteroids enhance airway epithelial barrier integrity. However, the mechanism by which they accomplish this is unclear. Therefore, we investigated steroid-inducible genes and signaling pathways that were involved in enhancing airway epithelial barrier integrity. Methods: A human bronchial epithelial cell line (16HBE cells) was cultured with 10−6 M dexamethasone (DEX) for 3 days to enhance epithelial barrier integrity. After measuring transepithelial electrical resistance (TER) and paracellular permeability, we extracted total RNA from 16HBE cells and performed microarray and pathway analysis. After we identified candidate genes and a canonical pathway, we measured TER and immunostained for tight junction (TJ) and adherent junction (AJ) proteins in cells that had been transfected with specific small interfering RNAs (siRNAs) for these genes. Results: We identified a nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated oxidative stress response pathway which was primarily involved in the steroid-induced enhancement of airway epithelial barrier integrity. Transfecting cells with Nrf2 specific siRNA reduced the steroid-induced enhancement of airway epithelial barrier integrity and the accumulation of TJ and AJ proteins at sites of cell–cell contact. Moreover, based on pathway analysis, aldehyde oxidase 1 (AOX1) was identified as a downstream enzyme of Nrf2. Transfecting cells with AOX1-specific siRNA also reduced the steroid-induced enhancement of airway epithelial barrier integrity. Conclusions: Our results indicated that the Nrf2/AOX1 pathway was important for enhancing airway epithelial barrier integrity. Because the airway epithelium of asthmatics is susceptible to reduced barrier integrity, this pathway might be a new therapeutic target for asthma.http://www.sciencedirect.com/science/article/pii/S1323893015001409AOX1AsthmaCorticosteroidNrf2Microarray analysis |
| spellingShingle | Yoshitaka Shintani Shuichiro Maruoka Yasuhiro Gon Daisuke Koyama Akiko Yoshida Yutaka Kozu Kazumichi Kuroda Ikuko Takeshita Eriko Tsuboi Kaori Soda Shu Hashimoto Nuclear factor erythroid 2-related factor 2 (Nrf2) regulates airway epithelial barrier integrity Allergology International AOX1 Asthma Corticosteroid Nrf2 Microarray analysis |
| title | Nuclear factor erythroid 2-related factor 2 (Nrf2) regulates airway epithelial barrier integrity |
| title_full | Nuclear factor erythroid 2-related factor 2 (Nrf2) regulates airway epithelial barrier integrity |
| title_fullStr | Nuclear factor erythroid 2-related factor 2 (Nrf2) regulates airway epithelial barrier integrity |
| title_full_unstemmed | Nuclear factor erythroid 2-related factor 2 (Nrf2) regulates airway epithelial barrier integrity |
| title_short | Nuclear factor erythroid 2-related factor 2 (Nrf2) regulates airway epithelial barrier integrity |
| title_sort | nuclear factor erythroid 2 related factor 2 nrf2 regulates airway epithelial barrier integrity |
| topic | AOX1 Asthma Corticosteroid Nrf2 Microarray analysis |
| url | http://www.sciencedirect.com/science/article/pii/S1323893015001409 |
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