Clarithromycin Suppresses Chloride Channel Accessory 1 and Inhibits Interleukin-13-Induced Goblet Cell Hyperplasia in Human Bronchial Epithelial Cells

Akimichi Nagashima; Masaharu Shinkai; Masahiro Shinoda; Tadasuke Shimokawaji; Yasuhiro Kimura; Kei Mishina; Takashi Sato; Mariko Toda; Yoshiaki Inayama; Bruce K Rubin; Takeshi Kaneko

doi:10.1128/AAC.01327-16

Clarithromycin Suppresses Chloride Channel Accessory 1 and Inhibits Interleukin-13-Induced Goblet Cell Hyperplasia in Human Bronchial Epithelial Cells

Antimicrob Agents Chemother. 2016 Oct 21;60(11):6585-6590. doi: 10.1128/AAC.01327-16. Print 2016 Nov.

Affiliations

¹ Respiratory Disease Center, Yokohama City University Medical Center, Yokohama, Japan.
² Respiratory Disease Center, Yokohama City University Medical Center, Yokohama, Japan [email protected].
³ Department of Pathology, Yokohama City University Medical Center, Yokohama, Japan.
⁴ Department of Pediatrics, Virginia Commonwealth University, Richmond, Virginia, USA.
⁵ Department of Pulmonology, Yokohama City University Graduate School of Medicine, Yokohama, Japan.

Abstract

Activation of the interleukin-13 (IL-13) receptor leads to signal transducer and activator of transcription 6 (STAT6) activation and subsequent induction of SAM pointed domain containing ETS transcription factor (SPDEF) and chloride channel accessory 1 (CLCA1), increasing secretion of the gel-forming mucin MUC5AC. Activation of the epidermal growth factor receptor (EGFR) also leads to MUC5AC production via extracellular signal-regulated kinase (ERK1/2). We examined the effect of clarithromycin IL-13 signaling leading to production. Normal human bronchial epithelial (NHBE) cells were grown for 14 days at an air-liquid interface (ALI) with IL-13 and/or clarithromycin. Histochemical analysis was performed using hematoxylin and eosin (HE) staining and MUC5AC immunostaining. MUC5AC, SPDEF, and CLCA1 mRNA expression were evaluated by real-time PCR. Western analysis was used to assess phosphorylation of STAT6 and ERK1/2. Clarithromycin decreased IL-13-induced goblet cell hyperplasia and MUC5AC mRNA expression in a dose-dependent manner. Clarithromycin decreased IL-13-stimulated SPDEF and CLCA1 mRNA expression in a dose-dependent manner, and at 32 μg/ml CLCA1 was profoundly decreased (P < 0.001). Although clarithromycin had no effect on STAT6 phosphorylation induced by IL-13, it decreased constitutive phosphorylation of ERK1/2 (P < 0.05).

MeSH terms

Bronchi / cytology
Bronchi / drug effects
Bronchi / metabolism
Cell Proliferation / drug effects
Cells, Cultured
Chloride Channels / antagonists & inhibitors
Chloride Channels / genetics*
Chloride Channels / metabolism
Clarithromycin / pharmacology*
Dose-Response Relationship, Drug
ErbB Receptors / genetics
ErbB Receptors / metabolism
Gene Expression Regulation
Goblet Cells / cytology
Goblet Cells / drug effects*
Goblet Cells / metabolism
Humans
Immunohistochemistry
Interleukin-13 / antagonists & inhibitors*
Interleukin-13 / pharmacology
Mitogen-Activated Protein Kinase 1 / genetics
Mitogen-Activated Protein Kinase 1 / metabolism
Mitogen-Activated Protein Kinase 3 / genetics
Mitogen-Activated Protein Kinase 3 / metabolism
Mucin 5AC / genetics
Mucin 5AC / metabolism
Phosphorylation / drug effects
Protein Synthesis Inhibitors / pharmacology*
Proto-Oncogene Proteins c-ets / genetics
Proto-Oncogene Proteins c-ets / metabolism
RNA, Messenger / genetics
RNA, Messenger / metabolism
Receptors, Interleukin-13 / genetics
Receptors, Interleukin-13 / metabolism
STAT6 Transcription Factor / genetics
STAT6 Transcription Factor / metabolism
Signal Transduction

Substances

CLCA1 protein, human
Chloride Channels
Interleukin-13
MUC5AC protein, human
Mucin 5AC
Protein Synthesis Inhibitors
Proto-Oncogene Proteins c-ets
RNA, Messenger
Receptors, Interleukin-13
SPDEF protein, human
STAT6 Transcription Factor
STAT6 protein, human
EGFR protein, human
ErbB Receptors
MAPK1 protein, human
Mitogen-Activated Protein Kinase 1
Mitogen-Activated Protein Kinase 3
Clarithromycin