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

Affiliations
  • 1Department of Otorhinolaryngology-Head and Neck Surgery, Yeungnam University College of Medicine, Gyeongsan, Korea. ydkim@med.yu.ac.kr
  • 2Department of Physiology, Yeungnam University College of Medicine, Gyeongsan, Korea.
  • 3Regional Center for Respiratory Diseases, Yeungnam University Medical Center, Daegu, Korea.

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.


Reference

1. Brownlee M. The pathobiology of diabetic complications: a unifying mechanism. Diabetes. 2005; Jun. 54(6):1615–25.
2. Baker EH, Wood DM, Brennan AL, Clark N, Baines DL, Philips BJ. Hyperglycaemia and pulmonary infection. Proc Nutr Soc. 2006; Aug. 65(3):227–35.
Article
3. Baker EH, Bell D. Blood glucose: of emerging importance in COPD exacerbations. Thorax. 2009; Oct. 64(10):830–2.
Article
4. Roy S, Trudeau K, Roy S, Tien T, Barrette KF. Mitochondrial dysfunction and endoplasmic reticulum stress in diabetic retinopathy: mechanistic insights into high glucose-induced retinal cell death. Curr Clin Pharmacol. 2013; Nov. 8(4):278–84.
Article
5. Wu D, Peng F, Zhang B, Ingram AJ, Kelly DJ, Gilbert RE, et al. EGFR-PLCgamma1 signaling mediates high glucose-induced PKCbeta1-Akt activation and collagen I upregulation in mesangial cells. Am J Physiol Renal Physiol. 2009; Sep. 297(3):F822–34.
6. Singh R, Kishore L, Kaur N. Diabetic peripheral neuropathy: current perspective and future directions. Pharmacol Res. 2014; Feb. 80:21–35.
Article
7. Lan CC, Wu CS, Huang SM, Wu IH, Chen GS. High-glucose environment enhanced oxidative stress and increased interleukin-8 secretion from keratinocytes: new insights into impaired diabetic wound healing. Diabetes. 2013; Jul. 62(7):2530–8.
8. Han L, Ma Q, Li J, Liu H, Li W, Ma G, et al. High glucose promotes pancreatic cancer cell proliferation via the induction of EGF expression and transactivation of EGFR. PLoS One. 2011. 6(11):e27074.
Article
9. Tiengo A, Fadini GP, Avogaro A. The metabolic syndrome, diabetes and lung dysfunction. Diabetes Metab. 2008; Nov. 34(5):447–54.
Article
10. Forgiarini LA Jr, Kretzmann NA, Porawski M, Dias AS, Marroni NA. Experimental diabetes mellitus: oxidative stress and changes in lung structure. J Bras Pneumol. 2009; Aug. 35(8):788–91.
11. Baker EH, Janaway CH, Philips BJ, Brennan AL, Baines DL, Wood DM, et al. Hyperglycaemia is associated with poor outcomes in patients admitted to hospital with acute exacerbations of chronic obstructive pulmonary disease. Thorax. 2006; Apr. 61(4):284–9.
Article
12. Yu H, Yang J, Xiao Q, Lu Y, Zhou X, Xia L, et al. Regulation of high glucose-mediated mucin expression by matrix metalloproteinase-9 in human airway epithelial cells. Exp Cell Res. 2015; Apr. 333(1):127–35.
Article
13. Bae CH, Choi YS, Song SY, Kim YD. Effect of thymic stromal lymphopoietin on MUC5B expression in human airway epithelial cells. Biochem Biophys Res Commun. 2014; May. 448(2):231–5.
Article
14. Kim YD, Kwon EJ, Park DW, Song SY, Yoon SK, Baek SH. Interleukin-1beta induces MUC2 and MUC5AC synthesis through cyclooxygenase-2 in NCI-H292 cells. Mol Pharmacol. 2002; Nov. 62(5):1112–8.
15. Jorissen RN, Walker F, Pouliot N, Garrett TP, Ward CW, Burgess AW. Epidermal growth factor receptor: mechanisms of activation and signalling. Exp Cell Res. 2003; Mar. 284(1):31–53.
Article
16. Burgel PR, Nadel JA. Epidermal growth factor receptor-mediated innate immune responses and their roles in airway diseases. Eur Respir J. 2008; Oct. 32(4):1068–81.
Article
17. Hao Y, Kuang Z, Jing J, Miao J, Mei LY, Lee RJ, et al. Mycoplasma pneumoniae modulates STAT3-STAT6/EGFR-FOXA2 signaling to induce overexpression of airway mucins. Infect Immun. 2014; Dec. 82(12):5246–55.
Article
18. Yuan-Chen Wu D, Wu R, Reddy SP, Lee YC, Chang MM. Distinctive epidermal growth factor receptor/extracellular regulated kinase-independent and -dependent signaling pathways in the induction of airway mucin 5B and mucin 5AC expression by phorbol 12-myristate 13-acetate. Am J Pathol. 2007; Jan. 170(1):20–32.
Article
19. Rada B, Gardina P, Myers TG, Leto TL. Reactive oxygen species mediate inflammatory cytokine release and EGFR-dependent mucin secretion in airway epithelial cells exposed to Pseudomonas pyocyanin. Mucosal Immunol. 2011; Mar. 4(2):158–71.
Article
20. Burgel PR, Montani D, Danel C, Dusser DJ, Nadel JA. A morphometric study of mucins and small airway plugging in cystic fibrosis. Thorax. 2007; Feb. 62(2):153–61.
Article
21. Puddicombe SM, Polosa R, Richter A, Krishna MT, Howarth PH, Holgate ST, et al. Involvement of the epidermal growth factor receptor in epithelial repair in asthma. FASEB J. 2000; Jul. 14(10):1362–74.
Article
22. Williams OW, Sharafkhaneh A, Kim V, Dickey BF, Evans CM. Airway mucus: from production to secretion. Am J Respir Cell Mol Biol. 2006; May. 34(5):527–36.
23. Rogers DF. Physiology of airway mucus secretion and pathophysiology of hypersecretion. Respir Care. 2007; Sep. 52(9):1134–46.
24. Rose MC, Voynow JA. Respiratory tract mucin genes and mucin glycoproteins in health and disease. Physiol Rev. 2006; Jan. 86(1):245–78.
Article
25. Turner J, Jones CE. Regulation of mucin expression in respiratory diseases. Biochem Soc Trans. 2009; Aug. 37(Pt 4):877–81.
Article
26. Woo HJ, Yoo WJ, Bae CH, Song SY, Kim YW, Park SY, et al. Leptin up-regulates MUC5B expression in human airway epithelial cells via mitogen-activated protein kinase pathway. Exp Lung Res. 2010; Jun. 36(5):262–9.
27. Ohtsu H, Dempsey PJ, Eguchi S. ADAMs as mediators of EGF receptor transactivation by G protein-coupled receptors. Am J Physiol Cell Physiol. 2006; Jul. 291(1):C1–10.
Article
28. Zwick E, Hackel PO, Prenzel N, Ullrich A. The EGF receptor as central transducer of heterologous signalling systems. Trends Pharmacol Sci. 1999; Oct. 20(10):408–12.
Article
Full Text Links
  • CEO
Actions
Cited
CITED
export Copy
Close
Share
  • Twitter
  • Facebook
Similar articles
Copyright © 2023 by Korean Association of Medical Journal Editors. All rights reserved.     E-mail: koreamed@kamje.or.kr