Tuberc Respir Dis.  2015 Jul;78(3):210-217. 10.4046/trd.2015.78.3.210.

Effects of Lupenone, Lupeol, and Taraxerol Derived from Adenophora triphylla on the Gene Expression and Production of Airway MUC5AC Mucin

• Yoon YP
• Lee HJ
• Lee DU
• Lee SK
• Hong JH
• Lee CJ

• KMID: 2320645
• DOI: http://doi.org/10.4046/trd.2015.78.3.210

Abstract

BACKGROUND
Adenophora triphylla var. japonica is empirically used for controlling airway inflammatory diseases in folk medicine. We evaluated the gene expression and production of mucin from airway epithelial cells in response to lupenone, lupeol and taraxerol derived from Adenophora triphylla var. japonica.
METHODS
Confluent NCI-H292 cells were pretreated with lupenone, lupeol or taraxerol for 30 minutes and then stimulated with tumor necrosis factor alpha (TNF-alpha) for 24 hours. The MUC5AC mucin gene expression and production were measured by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Additionally, we examined whether lupenone, lupeol or taraxerol affects MUC5AC mucin production induced by epidermal growth factor (EGF) and phorbol 12-myristate 13-acetate (PMA), the other 2 stimulators of airway mucin production.
RESULTS
Lupenone, lupeol, and taraxerol inhibited the gene expression and production of MUC5AC mucin induced by TNF-alpha from NCI-H292 cells, respectively. The 3 compounds inhibited the EGF or PMA-induced production of MUC5AC mucin in NCI-H292 cells.
CONCLUSION
These results indicated that lupenone, lupeol and taraxerol derived from Adenophora triphylla var. japonica regulates the production and gene expression of mucin, by directly acting on airway epithelial cells. In addition, the results partly explain the mechanism of of Adenophora triphylla var. japonica as a traditional remedy for diverse inflammatory pulmonary diseases.

Keyword

Mucin; Lupenone; Lupeol; Taraxerol

MeSH Terms

Campanulaceae*
Enzyme-Linked Immunosorbent Assay
Epidermal Growth Factor
Epithelial Cells
Gene Expression*
Lung Diseases
Medicine, Traditional
Methods
Mucins*
Tumor Necrosis Factor-alpha
Epidermal Growth Factor
Mucins
Tumor Necrosis Factor-alpha

Figure

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  Figure 1 Chemical structure of lupenone (A), lupeol (B), and taraxerol (C).

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  Figure 2 Effect of lupenone (A), lupeol (B), or taraxerol (C) on tumor necrosis factor α (TNF-α)-induced MUC5AC gene expression in NCI-H292 cells. NCI-H292 cells were pretreated with varying concentrations of lupenone, lupeol, or taraxerol for 30 minutes and then stimulated with TNF-α (0.2 nM) for 24 hours. MUC5AC gene expression was measured by reverse transcription-polymerase chain reaction. Three independent experiments were performed and the representative images were shown. Polymerase chain reaction images were quantified using densitometry and the graphic data were also shown. Each bar represents a mean±SEM of 3 densitograms in comparison with that of control set at 100% (A-C). *Significantly different from control (p<0.05). †Significantly different from TNF-α alone (p<0.05).

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  Figure 3 Effect of lupenone (A), lupeol (B), or taraxerol (C) on tumor necrosis factor α (TNF-α)-induced MUC5AC mucin production in NCI-H292 cells. NCI-H292 cells were pretreated with varying concentrations of lupenone, lupeol or taraxerol for 30 minutes and then stimulated with TNF-α (0.2 nM, 10 ng/mL) for 24 hours. Cell lysates were collected for measurement of MUC5AC mucin production by enzyme-linked immunosorbent assay. Each bar represents a mean±SEM of three culture wells in comparison with that of the control set at 100% (A-C). Three independent experiments were performed and the representative data were shown. *Significantly different from control (p<0.05). †Significantly different from TNF-α alone (p<0.05).

• 

  Figure 4 Effect of lupenone (A), lupeol (B), or taraxerol (C) on epidermal growth factor (EGF)-induced MUC5AC mucin production in NCI-H292 cells. NCI-H292 cells were pretreated with varying concentrations of lupenone, lupeol, or taraxerol for 30 minutes and then stimulated with EGF (25 ng/mL) for 24 hours. Cell lysates were collected for measurement of MUC5AC mucin production by enzyme-linked immunosorbent assay. Each bar represents a mean±SEM of three culture wells in comparison with that of control set at 100% (A-C). Three independent experiments were performed and the representative data were shown. *Significantly different from control (p<0.05). †Significantly different from EGF alone (p<0.05).

• 

  Figure 5 Effect of lupenone (A), lupeol (B), or taraxerol (C) on phorbol 12-myristate 13-acetate (PMA)-induced MUC5AC mucin production in NCI-H292 cells. NCI-H292 cells were pretreated with varying concentrations of lupenone, lupeol, or taraxerol for 30 minutes and then stimulated with PMA (10 ng/mL) for 24 hours. Cell lysates were collected for measurement of MUC5AC mucin production by enzyme-linked immunosorbent assay. Each bar represents a mean±SEM of three culture wells in comparison with that of control set at 100% (A-C). Three independent experiments were performed and the representative data were shown. *Significantly different from control (p<0.05). †Significantly different from PMA alone (p<0.05).

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