Tuberc Respir Dis.  2017 Jan;80(1):60-68. 10.4046/trd.2017.80.1.60.

Mometasone Furoate Suppresses PMA-Induced MUC-5AC and MUC-2 Production in Human Airway Epithelial Cells

• Poachanukoon O
• Koontongkaew S
• Monthanapisut P
• Pattanacharoenchai N

• KMID: 2396342
• DOI: http://doi.org/10.4046/trd.2017.80.1.60

Abstract

BACKGROUND
Mucus hypersecretion from airway epithelium is a characteristic feature of airway inflammatory diseases. Tumor necrosis factor α (TNF-α) regulates mucin synthesis. Glucocorticoids including mometasone fuorate (MF) have been used to attenuate airway inflammation. However, effects of MF on mucin production have not been reported.
METHODS
Effects of MF and budesonide (BUD) on the phorbol-12-myristate-13-acetate (PMA)-induction of mucin and TNF-α in human airway epithelial cells (NCI-H292) were investigated in the present study. Confluent NCI-H292 cells were pretreated with PMA (200 nM) for 2 hours. Subsequently, the cells were stimulated with MF (1-500 ng/mL) or BUD (21.5 ng/mL) for 8 hours. Dexamethasone (1 µg/mL) was used as the positive control. Real-time polymerase chain reaction was used to determine MUC2 and MUC5AC mRNA levels. The level of total mucin, MUC2, MUC5AC, and TNF-α in culture supernatants were measured using enzyme-linked immunosorbent assay.
RESULTS
MF and BUD significantly suppressed MUC2 and MUC5AC gene expression in PMA-stimulated NCI-H292 cells. The inhibitory effects of the two steroid drugs were also observed in the production of total mucin, MUC2 and MUC5AC proteins, and TNF-α.
CONCLUSION
Our findings demonstrated that MF and BUD attenuated mucin and TNF-α production in PMA-induced human airway epithelial cells.

Keyword

Mometasone Fuorate; Budesonide; Mucins; MUC2 Protein, Human; MUC5AC Protein, Human; Tumor Necrosis Factor-Alpha; Epithelial Cells

MeSH Terms

Budesonide
Dexamethasone
Enzyme-Linked Immunosorbent Assay
Epithelial Cells*
Epithelium
Gene Expression
Glucocorticoids
Humans*
Inflammation
Mometasone Furoate*
Mucins
Mucus
Real-Time Polymerase Chain Reaction
RNA, Messenger
Tumor Necrosis Factor-alpha
Budesonide
Dexamethasone
Glucocorticoids
Mometasone Furoate
Mucins
RNA, Messenger
Tumor Necrosis Factor-alpha

Figure

• 

  Figure 1 Effects of MF and BUD on cell viability. NCI-H292 cells were treated with MF (A) and BUD (B) at indicated concentrations for 72 hours. Cell viability was determined by the MTT assay and results are presented as percent of the control (untreated group). Values are expressed as mean±SEM of three independent experiments. MF: mometasone fuorate; BUD: budesonide; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; SEM: standard error of the mean.

• 

  Figure 2 Glucocorticoids inhibit PMA-induced MUC2 gene expression. NCI-H292 cells were pretreated with or without MF, BUD, or DEX at indicated concentrations for 2 hours. Subsequently, the cells were stimulated with PMA for 8 hours. Total RNA was isolated and MUC2 mRNA levels were analyzed by real-time polymerase chain reaction. Values are expressed as mean±SEM of three independent experiments. *p<0.05, compared to PMA-stimulated cells in the absence of glucocorticoids. PMA: phorbol-12-myristate-13-acetate; MF: mometasone fuorate; BUD: budesonide; DEX: dexamethasone; SEM: standard error of the mean; GAPDH: glyceraldehyde 3-phosphate dehydrogenase.

• 

  Figure 3 MF and BUD inhibit PMA-induced MUC5AC gene expression. NCI-H292 cell were pretreated with or without MF, BUD, or DEX at indicated concentrations for 2 hours before stimulation with PMA for 8 hours. Total RNA was isolated and MUC5AC mRNA levels were analyzed by real-time polymerase chain reaction. Values are expressed as mean±SEM of three independent experiments. *p<0.05, compared to PMA-stimulated cells in the absence of glucocorticoids. MF: mometasone fuorate; BUD: budesonide; PMA: phorbol-12-myristate-13-acetate; DEX: dexamethasone; SEM: standard error of the mean; GAPDH: glyceraldehyde 3-phosphate dehydrogenase.

• 

  Figure 4 MF and BUD inhibit PMA-induced MUC2 synthesis. NCI-H292 cells were pretreated with indicated glucocorticoids for 2 hours. Subsequently, the cells were stimulated with PMA for 8 hours. The level of MUC2 protein in culture supernatants was determined using ELISA. Data shown are mean±SEM of three independent experiments. *p<0.05, compared to PMA-stimulated cells in the absence of glucocorticoids. MF: mometasone fuorate; BUD: budesonide; PMA: phorbol-12-myristate-13-acetate; ELISA: enzyme-linked immunosorbent assay; SEM: standard error of the mean; DEX: dexamethasone.

• 

  Figure 5 MF and BUD inhibit PMA-induced MUC5AC production. NCI-H292 cells were pretreated with indicated glucocorticoids for 2 hours. Subsequently, the cells were stimulated with PMA for 8 hours. The level of MUC5AC protein in culture supernatants was determined using ELISA. Data shown are mean±SEM of three independent experiments. *p<0.05, compared to PMA-stimulated cells in the absence of glucocorticoids. MF: mometasone fuorate; BUD: budesonide; PMA: phorbol-12-myristate-13-acetate; ELISA: enzyme-linked immunosorbent assay; SEM: standard error of the mean; DEX: dexamethasone.

• 

  Figure 6 Inhibition of total mucin production by MF, BUD, and DEX. NCI-H292 cells were pretreated with the indicated glucocorticoids for 2 hours. The cells were stimulated with PMA and total mucin levels were quantitated by an enzyme-linked lectin assay. Data shown are mean±SEM of three independent experiments. *p<0.05, compared to PMA-stimulated cells in the absence of glucocorticoids. MF: mometasone fuorate; BUD: budesonide; DEX: dexamethasone; PMA: phorbol-12-myristate-13-acetate; SEM: standard error of the mean.

• 

  Figure 7 Glucocorticoids suppress the PMA-induced release of TNF-α from NCI-H292 cells. NCI-H292 cells were pretreated with MF, BUD, or DEX for 2 hours before the cells were stimulated with PMA for 8 hours. TNF-α in the culture supernatant was quantitated by ELISA. Data shown are mean±SEM of three independent experiments. *p<0.05, compared to PMA-stimulated cells in the absence of glucocorticoids. PMA: phorbol-12-myristate-13-acetate; TNF-α: tumor necrosis factor α; MF: mometasone fuorate; BUD: budesonide; DEX: dexamethasone; ELISA: enzyme-linked immunosorbent assay; SEM: standard error of the mean.

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