HO-1 inhibits IL-13-induced goblet cell hyperplasia associated with CLCA1 suppression in normal human bronchial epithelial cells.
Mucus hypersecretion and goblet cell hyperplasia are common features that characterize asthma. IL-13 increases mucin (MUC) 5AC, the major component of airway mucus, in airway epithelial cells. According to the literature, IL-13 receptor activation leads to STAT6 activation and consequent induction of chloride channel accessory 1 (CLCA1) gene expression, associated with the induction of MUC5AC. Heme oxygenase-1 (HO-1) is an enzyme that catalyzes oxidation of heme to biliverdin, and has anti-inflammatory and anti-oxidant properties. We examined the effects of HO-1 on mucin production and goblet cell hyperplasia induced by IL-13. Moreover, we assessed the cell signaling intermediates that appear to be responsible for mucin production. Normal human bronchial epithelial (NHBE) cells were grown at air liquid interface (ALI) in the presence or absence of IL-13 and hemin, a HO-1 inducer, for 14 days. Protein concentration was analyzed using ELISA, and mRNA expression was examined by real-time PCR. Histochemical analysis was performed using HE staining, andWestern blotting was performed to evaluate signaling transduction pathway. Hemin (4 μM) significantly increased HO-1 protein expression (p b 0.01) and HO-1 mRNA expression (p b 0.001). IL-13 significantly increased goblet cells, MUC5AC protein secretion (p b 0.01) and MUC5AC mRNA (p b 0.001), and these were decreased by hemin by way of HO-1. Tin protoporphyrin (SnPP)-IX, a HO-1 inhibitor, blocked the effect of hemin restoring MUC5AC protein secretion (p b 0.05) and goblet cell hyperplasia. Hemin decreased the expression of CLCA1 mRNA (p b 0.05) and it was reversed by SnPP-IX, but could not suppress IL-13-induced phosphorylation of STAT6 or SAM pointed domain-containing ETS transcription factor (SPDEF) and Forkhead box A2 (FOXA2) mRNA expression. In summary, HO-1 overexpression suppressed IL-13-induced goblet cell hyperplasia and MUC5AC production, and involvement of CLCA1 in the mechanism was suggested.
Respiratory Disease Center, Yokohama City University Medical Center, Yokohama, Japan.
Kei
K
Mishina
Respiratory Disease Center, Yokohama City University Medical Center, Yokohama, Japan. Electronic address: shinkai@yokohama-cu.ac.jp.
Masaharu
M
Shinkai
Respiratory Disease Center, Yokohama City University Medical Center, Yokohama, Japan.
Tadasuke
T
Shimokawaji
Respiratory Disease Center, Yokohama City University Medical Center, Yokohama, Japan.
Akimichi
A
Nagashima
Respiratory Disease Center, Yokohama City University Medical Center, Yokohama, Japan.
Yusuke
Y
Hashimoto
Respiratory Disease Center, Yokohama City University Medical Center, Yokohama, Japan.
Yoriko
Y
Inoue
Department of Pathology, Yokohama City University Medical Center, Yokohama, Japan.
Yoshiaki
Y
Inayama
Department of Pediatrics, Virginia Commonwealth University, Richmond, USA.
Bruce K
BK
Rubin
Department of Internal Medicine and Clinical Immunology, Yokohama City University Graduate School of Medicine, Yokohama, Japan.
Yoshiaki
Y
Ishigatsubo
Department of Pulmonology, Yokohama City University Graduate School of Medicine, Yokohama, Japan.
Takeshi
T
Kaneko
D001980:Bronchi / Q000166:cytology / Q000187:drug effects; D002478:Cells, Cultured; D018118:Chloride Channels / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000187:drug effects*; D020397:Goblet Cells / Q000187:drug effects*; D051547:Heme Oxygenase-1 / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D006427:Hemin / Q000378:metabolism; D006801:Humans; D006965:Hyperplasia / Q000139:chemically induced / Q000517:prevention & control*; D018793:Interleukin-13 / Q000037:antagonists & inhibitors* / Q000494:pharmacology*; D008665:Metalloporphyrins / Q000494:pharmacology; D055271:Mucin 5AC / Q000096:biosynthesis; D011524:Protoporphyrins / Q000494:pharmacology; D020545:Respiratory Mucosa / Q000166:cytology / Q000187:drug effects; D050800:STAT6 Transcription Factor / Q000037:antagonists & inhibitors / Q000096:biosynthesis
C492109:CLCA1 protein, human; D018118:Chloride Channels; D004791:Enzyme Inhibitors; D018793:Interleukin-13; C524678:MUC5AC protein, human; D008665:Metalloporphyrins; D055271:Mucin 5AC; D011524:Protoporphyrins; D050800:STAT6 Transcription Factor; C494101:STAT6 protein, human; D006427:Hemin; C032628:tin protoporphyrin IX; D051547:Heme Oxygenase-1
CLCA1 [D12.776.157.530.400.175]; Goblet cell hyperplasia [A04.760.259]; HO-1 [D08.811.682.690.708.410.500]; IL-13 [D12.644.276.374.465.513]; MUC5AC [D12.776.395.560.631.300.500]; Normal human bronchial epithelial cells (respiratory mucosa) [A04.760]