Tuberc Respir Dis.  2010 Nov;69(5):348-353.

Regulation of Tumor Necrosis Factor-alpha-induced Airway Mucin Production and Gene Expression by Carbenoxolone, Prunetin, and Silibinin

• Lee HJ
• Lee SY
• Jeon BK
• Lee JW
• Lee MN
• Kim JO
• Lee CJ

Abstract

BACKGROUND
In this study, we tried to investigate whether carbenoxolone, prunetin, and silibinin affect tumor necrosis factor (TNF)-alpha-induced MUC5AC mucin production and gene expression from human airway epithelial cells.
METHODS
Confluent NCI-H292 cells were pretreated with each agent (carbenoxolone, prunetin, and silibinin) for 30 min and then stimulated with TNF-alpha for 24 hours. The MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assay, respectively.
RESULTS
Carbenoxolone, prunetin and silibinin inhibited the production of MUC5AC mucin protein induced by TNF-alpha; the 3 compounds also inhibited the expression of MUC5AC mucin gene induced by TNF-alpha.
CONCLUSION
This result suggests that carbenoxolone, prunetin and silibinin can inhibit mucin gene expression and production of mucin protein induced by TNF-alpha, by directly acting on airway epithelial cells.

Keyword

MUC5AC protein, human; Carbenoxolone; Prunetin; Silibinin

MeSH Terms

Carbenoxolone
Enzyme-Linked Immunosorbent Assay
Epithelial Cells
Gene Expression
Humans
Isoflavones
Mucin 5AC
Mucins
Necrosis
Silymarin
Tumor Necrosis Factor-alpha
Carbenoxolone
Isoflavones
Mucin 5AC
Mucins
Silymarin
Tumor Necrosis Factor-alpha

Figure

• 

  Figure 1 Effect of carbenoxolone on tumor necrosis factor (TNF)-α-induced MUC5AC mucin production from NCI-H292 cells. NCI-H292 cells were pretreated with various concentrations of carbenoxolone for 30 minutes and then stimulated with TNF-α (0.2 nM) for 24 hours. Cell lysates were collected for measurement of MUC5AC mucin production by enzyme-linked immunosorbent assay. Means of individual group were converted to percent control and expressed as mean±SEM. The difference between groups was assessed using one-way ANOVA and Student's t-test for unpaired samples. p<0.05 was considered as significantly different. Each bar represents a mean±SEM of 3~4 culture wells in comparison with that of control set at 100%. con: control; Car: carbenoxolone. *Significantly different from control (p<0.05), †Significantly different from TNF-α alone (p<0.05).

• 

  Figure 2 Effect of prunetin on tumor necrosis factor (TNF)-α-induced MUC5AC mucin production from NCI-H292 cells. NCI-H292 cells were pretreated with various concentrations of prunetin for 30 minutes and then stimulated with TNF-α (0.2 nM) for 24 hours. Cell lysates were collected for measurement of MUC5AC mucin production by enzyme-linked immunosorbent assay. Means of individual group were converted to percent control and expressed as mean±SEM. The difference between groups was assessed using one-way ANOVA and Student's t-test for unpaired samples. p<0.05 was considered as significantly different. Each bar represents a mean±SEM of 3~4 culture wells in comparison with that of control set at 100%. cont: control, Pru: prunetin. *Significantly different from control (p<0.05), †Significantly different from TNF-α alone (p<0.05).

• 

  Figure 3 Effect of silibinin on tumor necrosis factor (TNF)-α-induced MUC5AC mucin production from NCI-H292 cells. NCI-H292 cells were pretreated with various concentrations of silibinin for 30 minutes and then stimulated with TNF-α (0.2 nM) for 24 hours. Cell lysates were collected for measurement of MUC5AC mucin production by enzyme-linked immunosorbent assay. Means of individual group were converted to percent control and expressed as mean±SEM. The difference between groups was assessed using one-way ANOVA and Student's t-test for unpaired samples. p<0.05 was considered as significantly different. Each bar represents a mean±SEM of 3~4 culture wells in comparison with that of control set at 100%. cont: control; Sil: silibinin. *Significantly different from control (p<0.05), †Significantly different from TNF-α alone (p<0.05).

• 

  Figure 4 Effects of carbenoxolone, prunetin and silibinin on tumor necrosis factor (TNF)-α-induced MUC5AC mucin gene expression from NCI-H292 cells. NCI-H292 cells were pretreated with carbenoxolone, prunetin, and silibinin (100 µM), respectively, 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. cont: control; Car: carbenoxolone; Pru: prunetin; Sil: silibinin.

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