PMID- 31706310 OWN - NLM STAT- MEDLINE DCOM- 20200427 LR - 20221207 IS - 1465-993X (Electronic) IS - 1465-9921 (Print) IS - 1465-9921 (Linking) VI - 20 IP - 1 DP - 2019 Nov 9 TI - Effects of cigarette smoke on barrier function and tight junction proteins in the bronchial epithelium: protective role of cathelicidin LL-37. PG - 251 LID - 10.1186/s12931-019-1226-4 [doi] LID - 251 AB - BACKGROUND: Airway epithelial barrier function is maintained by the formation of tight junctions (TJs) and adherens junctions (AJs). Inhalation of cigarette smoke causes airway epithelial barrier dysfunction and may contribute to the pathogenesis of chronic lung diseases such as asthma and chronic obstructive pulmonary disease (COPD). We assessed the effects of cigarette smoke on barrier function and expression of multiple TJ and AJ proteins in the bronchial epithelium. We also examined whether treatment with glucocorticosteroids (GCSs), long-acting beta(2)-agonists (LABAs), and human cathelicidin LL-37 can protect against cigarette smoke extract (CSE)-induced barrier dysfunction. METHODS: Calu-3 cells cultured at the air-liquid interface were pretreated with or without GCSs, LABAs, GCSs plus LABAs, or LL-37, and subsequently exposed to CSE. Barrier function was assessed by transepithelial electronic resistance (TEER) measurements. Gene and protein expression levels of TJ and AJ proteins were analyzed by quantitative PCR and western blotting, respectively. Immunofluorescence staining of TJ and AJ proteins was performed. RESULTS: CSE decreased TEER and increased permeability in a concentration-dependent manner. CSE suppressed gene expression of claudin-1, claudin-3, claudin-4, claudin-7, claudin-15, occludin, E-cadherin, junctional adhesion molecule-A (JAM-A) and zonula occludens-1 (ZO-1) within 12 h post-CSE exposure, while suppressed protein expression levels of occludin at 12 h. CSE-treated cells exhibited discontinuous or attenuated immunostaining for claudin-1, claudin-3, claudin-4, occludin, ZO-1, and E-cadherin compared with untreated cells. GCS treatment partially restored CSE-induced TEER reduction, while LABA treatment had no effect. GCS and LABA combination treatment had no additive effect on CSE-induced TEER reduction and gene suppression of TJ and AJ proteins. Human cathelicidin LL-37 counteracted CSE-induced TEER reduction and prevented disruption of occludin and ZO-1. LL-37 also attenuated CSE-induced decreases in gene and protein expression levels of occludin. CONCLUSIONS: CSE caused airway epithelial barrier dysfunction and simultaneously downregulated multiple TJ and AJ proteins. GCS and LABA combination treatment had no additive effect on CSE-induced TEER reduction. LL-37 counteracted CSE-induced TEER reduction and prevented disruption of occludin and ZO-1. Use of LL-37 to counteract airway epithelial barrier dysfunction may have significant benefits for respiratory diseases such as asthma and COPD. FAU - Tatsuta, Miyoko AU - Tatsuta M AD - Research Institute for Diseases of the Chest, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. AD - Department of Respiratory Medicine, National Hospital Organization Omuta National Hospital, Fukuoka, 837-0911, Japan. FAU - Kan-O, Keiko AU - Kan-O K AUID- ORCID: 0000-0002-7736-588X AD - Research Institute for Diseases of the Chest, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. hanamura@kokyu.med.kyushu-u.ac.jp. AD - Department of Endoscopic Diagnostics and Therapeutics, Kyushu University Hospital, Fukuoka, 812-8582, Japan. hanamura@kokyu.med.kyushu-u.ac.jp. FAU - Ishii, Yumiko AU - Ishii Y AD - Research Institute for Diseases of the Chest, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. FAU - Yamamoto, Norio AU - Yamamoto N AD - Research Institute for Diseases of the Chest, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. FAU - Ogawa, Tomohiro AU - Ogawa T AD - Research Institute for Diseases of the Chest, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. FAU - Fukuyama, Satoru AU - Fukuyama S AD - Research Institute for Diseases of the Chest, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. FAU - Ogawa, Aimi AU - Ogawa A AD - Research Institute for Diseases of the Chest, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. FAU - Fujita, Akitaka AU - Fujita A AD - Research Institute for Diseases of the Chest, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. FAU - Nakanishi, Yoichi AU - Nakanishi Y AD - Research Institute for Diseases of the Chest, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. FAU - Matsumoto, Koichiro AU - Matsumoto K AD - Research Institute for Diseases of the Chest, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan. LA - eng PT - Journal Article DEP - 20191109 PL - England TA - Respir Res JT - Respiratory research JID - 101090633 RN - 0 (Antimicrobial Cationic Peptides) RN - 0 (Smoke) RN - 0 (Tight Junction Proteins) RN - 0 (Cathelicidins) SB - IM MH - Antimicrobial Cationic Peptides/*pharmacology MH - Bronchi/*drug effects/metabolism MH - Cell Line MH - Electric Impedance MH - Epithelial Cells/*drug effects/metabolism MH - Gene Expression Regulation MH - Humans MH - Permeability MH - Signal Transduction MH - Smoke/*adverse effects MH - Tight Junction Proteins/genetics/*metabolism MH - Tight Junctions/*drug effects/genetics/metabolism MH - Tobacco Products/*adverse effects MH - Cathelicidins PMC - PMC6842552 OTO - NOTNLM OT - Airway epithelial barrier function OT - Cathelicidin OT - Cigarette smoke OT - Glucocorticosteroid OT - LL-37 OT - Long-acting beta2-agonist COIS- KK received a research grant from MSD Life Science Foundation, Public Interest Incorporated Foundation that was unrelated to the submitted work. KM received honorarium for educational lectures from AstraZeneca. The other authors declare that they have no competing interests. EDAT- 2019/11/11 06:00 MHDA- 2020/04/28 06:00 PMCR- 2019/11/09 CRDT- 2019/11/11 06:00 PHST- 2019/06/03 00:00 [received] PHST- 2019/10/31 00:00 [accepted] PHST- 2019/11/11 06:00 [entrez] PHST- 2019/11/11 06:00 [pubmed] PHST- 2020/04/28 06:00 [medline] PHST- 2019/11/09 00:00 [pmc-release] AID - 10.1186/s12931-019-1226-4 [pii] AID - 1226 [pii] AID - 10.1186/s12931-019-1226-4 [doi] PST - epublish SO - Respir Res. 2019 Nov 9;20(1):251. doi: 10.1186/s12931-019-1226-4.