Clin Exp Otorhinolaryngol.  2017 Mar;10(1):77-84. https://doi.org/10.21053/ceo.2016.00045.

Effect of High Glucose on MUC5B Expression in Human Airway Epithelial Cells

• Ye SB
• Choi YS
• Choi YH
• Bae CH
• Kim YW
• Park SY
• Song SY
• Kim YD

• KMID: 2376412
• DOI: http://doi.org/https://doi.org/10.21053/ceo.2016.00045

Abstract

OBJECTIVES
Excessive production of mucus results in plugging of the airway tract, which can increase morbidity and mortality in affected patients. In patients with diabetes, inflammatory airway disease appears with more frequent relapse and longer duration of symptoms. However, the effects of high glucose (HG) on the secretion of mucin in inflammatory respiratory diseases are not clear. Therefore, this study was conducted in order to investigate the effect and the brief signaling pathway of HG on MUC5B expression in human airway epithelial cells.
METHODS
The effect and signaling pathway of HG on MUC5B expression were investigated using reverse transcriptase-polymerase chain reaction (RT-PCR), real-time PCR, enzyme immunoassay, and immunoblot analysis with specific inhibitors and small interfering RNA.
RESULTS
HG increased MUC5B expression and epidermal growth factor receptor (EGFR) expression, and activated the phosphorylation of EGFR and p38 mitogen-activated protein kinase (MAPK). Pretreatment with EGFR inhibitor significantly attenuated the HG-induced phosphorylation of p38 MAPK, and pretreatments with p38 inhibitor or EGFR inhibitor significantly attenuated HG-induced MUC5B expression. In addition, knockdown of p38 MAPK by p38 MAPK siRNA significantly blocked HG-induced MUC5B expression.
CONCLUSION
These findings suggest that HG induces MUC5B expression via the sequential activations of the EGFR/p38 MAPK signaling pathway in human airway epithelial cells.

Keyword

Glucose; Receptor, Epidermal Growth Factor; MUC5B Protein; p38 Mitogen-Activated Protein Kinases; Epithelial Cell

MeSH Terms

Epithelial Cells*
Glucose*
Humans*
Immunoenzyme Techniques
Mortality
Mucins
Mucus
p38 Mitogen-Activated Protein Kinases
Phosphorylation
Protein Kinases
Real-Time Polymerase Chain Reaction
Receptor, Epidermal Growth Factor
Recurrence
RNA, Small Interfering
Glucose
Mucins
Protein Kinases
RNA, Small Interfering
Receptor, Epidermal Growth Factor
p38 Mitogen-Activated Protein Kinases

Figure

• 

  Fig. 1. Effect of high glucose (HG) on the expression of secretary airway mucins in NCI-H292 cells. Reverse transcriptase-polymerase chain reaction showed that HG (>15 mM) significantly induced MUC5B mRNA expression. However, glucose did not induce MUC2 or MUC5AC mRNA expression at any concentration examined. Images are representative of three separate experiments performed in triplicate. Bars indicate the mean±standard deviation of three independent experiments performed in triplicate. *P<0.05.

• 

  Fig. 2. Effect of high glucose (HG) on MUC5B expression in NCI-H292 cells. Real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) showed that MUC5B expression was significantly increased at concentrations higher than 15 mM, compared with the control group (5 mM). However, glucose did not induced MUC2 and MUC5AC expression (A, B). Real-time PCR showed that MUC5B mRNA expression was significantly increased at all times after exposure of glucose (20 mM) (C). Images are representative of three separate experiments performed in triplicate. Bars indicate the mean±standard deviation of three independent experiments performed in triplicate. *P<0.05.

• 

  Fig. 3. Effect of high glucose (HG) on the activation of epidermal growth factor receptor (EGFR) in NCI-H292 cells. Real-time polymerase chain reaction showed that EGFR mRNA expression was significantly increased at concentrations higher than 15 mM and peaked at 20 mM of glucose, compared with the control group (5 mM) (A). Western blot showed that HG significantly activated the phosphorylation of EGFR with the lapse of time (B). Images are representative of three separate experiments performed in triplicate. Bars indicate the mean±standard deviation of three independent experiments performed in triplicate. *P<0.05.

• 

  Fig. 4. Roles of epidermal growth factor receptor (EGFR), ERK1/2 mitogen-activated protein kinase (MAPK), and p38 MAPK on high glucose (HG)-induced MUC5B expression. (A) Western blot showed that HG significantly activated the phosphorylation of p38 MAPK with the lapse of time, but that HG did not activate the phosphorylation of ERK1/2 MAPK. (B, C) Reverse transcriptase-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) showed that SB203580 (a p38 MAPK inhibitor) and AG1478 (an EGFR inhibitor) significantly attenuated HG-induced MUC5B expression, whereas U0126 (an ERK1/2 inhibitor) had no effect. (D) Western blot showed that AG1478 significantly attenuated the HG-induced phosphorylation of p38 MAPK. (E and F) RT-PCR and ELISA showed that knockdown of p38 MAPK by p38 MAPK siRNA significantly blocked HG-induced MUC5B expression. Images are representative of three separate experiments performed in triplicate. Bars indicate the mean±standard deviation of three independent experiments performed in triplicate. *P<0.05.

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