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

Affiliations
  • 1Department of Pharmacology, Chungnam National University School of Medicine, Daejeon, Korea. LCJ123@cnu.ac.kr
  • 2Department of Radiologic Technology, Daegu Health College, Daegu, Korea.
  • 3LG Life Science, Seoul, Korea.
  • 4Pulmonology Section, Department of Internal Medicine, Chungnam National University Hospital, Chungnam National University School of Medicine, Daejeon, Korea.

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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