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Apigenin Inhibits Tumor Necrosis Factor-α-Induced Production and Gene Expression of Mucin through Regulating Nuclear Factor-Kappa B Signaling Pathway in Airway Epithelial Cells.

Seo HS, Sikder MA, Lee HJ, Ryu J, Lee CJ - Biomol Ther (Seoul) (2014)

Bottom Line: Apigenin significantly inhibited MUC5AC mucin production and down-regulated MUC5AC gene expression induced by TNF-α in NCI-H292 cells.To elucidate the action mechanism of apigenin, effect of apigenin on TNF-α-induced nuclear factor kappa B (NF-κB) signaling pathway was also investigated by western blot analysis.Apigenin also has an influence on upstream signaling of IKK because it inhibited the expression of adaptor protein, receptor interacting protein 1 (RIP1).

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, School of Medicine, Chungnam National University, Daejeon 301-131, Republic of Korea.

ABSTRACT
In the present study, we investigated whether apigenin significantly affects tumor necrosis factor-α (TNF-α)-induced production and gene expression of MUC5AC mucin in airway epithelial cells. Confluent NCI-H292 cells were pretreated with apigenin for 30 min and then stimulated with TNF-α for 24 h or the indicated periods. The MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription - polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. Apigenin significantly inhibited MUC5AC mucin production and down-regulated MUC5AC gene expression induced by TNF-α in NCI-H292 cells. To elucidate the action mechanism of apigenin, effect of apigenin on TNF-α-induced nuclear factor kappa B (NF-κB) signaling pathway was also investigated by western blot analysis. Apigenin inhibited NF-κB activation induced by TNF-α. Inhibition of inhibitory kappa B kinase (IKK) by apigenin led to the suppression of inhibitory kappa B alpha (IκBα) phosphorylation and degradation, p65 nuclear translocation. This, in turn, led to the down-regulation of MUC5AC protein production in NCI-H292 cells. Apigenin also has an influence on upstream signaling of IKK because it inhibited the expression of adaptor protein, receptor interacting protein 1 (RIP1). These results suggest that apigenin can regulate the production and gene expression of mucin through regulating NF-κB signaling pathway in airway epithelial cells.

No MeSH data available.


Related in: MedlinePlus

Effect of apigenin on TNF-α-induced MUC5AC mucin production and gene expression. NCI-H292 cells were pretreated with various concentrations of apigenin (5, 10 and 20 μM) for 30 min and then stimulated with TNF-α (10 ng/mL) for 24 h. Cell lysates were collected for measurement of MUC5AC mucin production by ELISA. Three independent experiments were performed and the representative data were shown. Each bar represents a mean ± S.E.M. of 3 culture wells in comparison with that of control set at 100% (A). MUC5AC mucin gene expression was measured by RT-PCR. As quantitative control, Rig/S15 rRNA, which encodes a small ribosomal subunit protein, a housekeeping gene that was constitutively expressed, was used. Three independent experiments were performed and the representative data were shown (B). *significantly different from control (p<0.05). †significantly different from TNF-α alone (p<0.05). (cont: control, Api: apigenin, TNF: TNF-α, concentration unit is μM).
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f1-bt-22-525: Effect of apigenin on TNF-α-induced MUC5AC mucin production and gene expression. NCI-H292 cells were pretreated with various concentrations of apigenin (5, 10 and 20 μM) for 30 min and then stimulated with TNF-α (10 ng/mL) for 24 h. Cell lysates were collected for measurement of MUC5AC mucin production by ELISA. Three independent experiments were performed and the representative data were shown. Each bar represents a mean ± S.E.M. of 3 culture wells in comparison with that of control set at 100% (A). MUC5AC mucin gene expression was measured by RT-PCR. As quantitative control, Rig/S15 rRNA, which encodes a small ribosomal subunit protein, a housekeeping gene that was constitutively expressed, was used. Three independent experiments were performed and the representative data were shown (B). *significantly different from control (p<0.05). †significantly different from TNF-α alone (p<0.05). (cont: control, Api: apigenin, TNF: TNF-α, concentration unit is μM).

Mentions: Apigenin inhibited TNF-α-induced MUC5AC mucin production. The amounts of MUC5AC mucin in the cells of apigenintreated cultures were 100 ± 8%, 368 ± 28%, 225 ± 13%, 153 ± 13% and 98 ± 8% for control, TNF-α 10 ng/mL only, TNF-α plus apigenin 5 μM, TNF-α plus apigenin 10 μM and TNF-α plus apigenin 20 μM, respectively (Fig. 1A). MUC5AC gene expression induced by TNF-α was also inhibited by pretreatment with 10 μM and 20 μM of apigenin (Fig. 1B). Cell viability was checked by sulforhodamine B (SRB) assay and there was no cytotoxic effect of apigenin, at 5, 10 and 20 μM concentration (data were not shown).


Apigenin Inhibits Tumor Necrosis Factor-α-Induced Production and Gene Expression of Mucin through Regulating Nuclear Factor-Kappa B Signaling Pathway in Airway Epithelial Cells.

Seo HS, Sikder MA, Lee HJ, Ryu J, Lee CJ - Biomol Ther (Seoul) (2014)

Effect of apigenin on TNF-α-induced MUC5AC mucin production and gene expression. NCI-H292 cells were pretreated with various concentrations of apigenin (5, 10 and 20 μM) for 30 min and then stimulated with TNF-α (10 ng/mL) for 24 h. Cell lysates were collected for measurement of MUC5AC mucin production by ELISA. Three independent experiments were performed and the representative data were shown. Each bar represents a mean ± S.E.M. of 3 culture wells in comparison with that of control set at 100% (A). MUC5AC mucin gene expression was measured by RT-PCR. As quantitative control, Rig/S15 rRNA, which encodes a small ribosomal subunit protein, a housekeeping gene that was constitutively expressed, was used. Three independent experiments were performed and the representative data were shown (B). *significantly different from control (p<0.05). †significantly different from TNF-α alone (p<0.05). (cont: control, Api: apigenin, TNF: TNF-α, concentration unit is μM).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4256032&req=5

f1-bt-22-525: Effect of apigenin on TNF-α-induced MUC5AC mucin production and gene expression. NCI-H292 cells were pretreated with various concentrations of apigenin (5, 10 and 20 μM) for 30 min and then stimulated with TNF-α (10 ng/mL) for 24 h. Cell lysates were collected for measurement of MUC5AC mucin production by ELISA. Three independent experiments were performed and the representative data were shown. Each bar represents a mean ± S.E.M. of 3 culture wells in comparison with that of control set at 100% (A). MUC5AC mucin gene expression was measured by RT-PCR. As quantitative control, Rig/S15 rRNA, which encodes a small ribosomal subunit protein, a housekeeping gene that was constitutively expressed, was used. Three independent experiments were performed and the representative data were shown (B). *significantly different from control (p<0.05). †significantly different from TNF-α alone (p<0.05). (cont: control, Api: apigenin, TNF: TNF-α, concentration unit is μM).
Mentions: Apigenin inhibited TNF-α-induced MUC5AC mucin production. The amounts of MUC5AC mucin in the cells of apigenintreated cultures were 100 ± 8%, 368 ± 28%, 225 ± 13%, 153 ± 13% and 98 ± 8% for control, TNF-α 10 ng/mL only, TNF-α plus apigenin 5 μM, TNF-α plus apigenin 10 μM and TNF-α plus apigenin 20 μM, respectively (Fig. 1A). MUC5AC gene expression induced by TNF-α was also inhibited by pretreatment with 10 μM and 20 μM of apigenin (Fig. 1B). Cell viability was checked by sulforhodamine B (SRB) assay and there was no cytotoxic effect of apigenin, at 5, 10 and 20 μM concentration (data were not shown).

Bottom Line: Apigenin significantly inhibited MUC5AC mucin production and down-regulated MUC5AC gene expression induced by TNF-α in NCI-H292 cells.To elucidate the action mechanism of apigenin, effect of apigenin on TNF-α-induced nuclear factor kappa B (NF-κB) signaling pathway was also investigated by western blot analysis.Apigenin also has an influence on upstream signaling of IKK because it inhibited the expression of adaptor protein, receptor interacting protein 1 (RIP1).

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, School of Medicine, Chungnam National University, Daejeon 301-131, Republic of Korea.

ABSTRACT
In the present study, we investigated whether apigenin significantly affects tumor necrosis factor-α (TNF-α)-induced production and gene expression of MUC5AC mucin in airway epithelial cells. Confluent NCI-H292 cells were pretreated with apigenin for 30 min and then stimulated with TNF-α for 24 h or the indicated periods. The MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription - polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. Apigenin significantly inhibited MUC5AC mucin production and down-regulated MUC5AC gene expression induced by TNF-α in NCI-H292 cells. To elucidate the action mechanism of apigenin, effect of apigenin on TNF-α-induced nuclear factor kappa B (NF-κB) signaling pathway was also investigated by western blot analysis. Apigenin inhibited NF-κB activation induced by TNF-α. Inhibition of inhibitory kappa B kinase (IKK) by apigenin led to the suppression of inhibitory kappa B alpha (IκBα) phosphorylation and degradation, p65 nuclear translocation. This, in turn, led to the down-regulation of MUC5AC protein production in NCI-H292 cells. Apigenin also has an influence on upstream signaling of IKK because it inhibited the expression of adaptor protein, receptor interacting protein 1 (RIP1). These results suggest that apigenin can regulate the production and gene expression of mucin through regulating NF-κB signaling pathway in airway epithelial cells.

No MeSH data available.


Related in: MedlinePlus