Choong Jae Lee

Genistein and curcumin suppress epidermal growth factor-induced MUC5AC mucin production and gene expression from human airway epithelial cells.

This study investigated whether genistein and curcumin affect epidermal growth factor (EGF)‐induced MUC5AC mucin production and gene expression from human airway epithelial cells. Confluent NCI‐H292 cells were pretreated with each agent for 30 min and then stimulated with EGF for 24 h. 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. The results were as follows: (1) genistein and curcumin inhibited the production of MUC5AC mucin protein induced by EGF, dose‐dependently; (2) genistein and curcumin also inhibited the expression of MUC5AC mucin gene induced by EGF. This result suggests that genistein and curcumin can regulate mucin gene expression and production of mucin protein induced by EGF, by directly acting on airway epithelial cells. Copyright

Inhibition of secretion, production and gene expression of mucin from cultured airway epithelial cells by prunetin.

This study investigated whether prunetin significantly affects the secretion, production and gene expression of mucin from cultured airway epithelial cells. Confluent primary rat tracheal surface epithelial (RTSE) cells were pretreated with adenosine triphosphate (ATP) for 5 min and then chased for 30 min in the presence of prunetin to assess the effect on mucin secretion using enzyme‐linked immunosorbent assay (ELISA). At the same time, confluent NCI‐H292 cells were pretreated with prunetin for 30 min and then stimulated with epidermal growth factor (EGF) or phorbol 12‐myristate 13‐acetate (PMA) for 24 h, respectively. The MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription‐polymerase chain reaction (RT‐PCR) and ELISA. The results were as follows: (1) prunetin significantly suppressed ATP‐induced mucin secretion from cultured RTSE cells; (2) prunetin inhibited the production of MUC5AC mucin protein induced by EGF or PMA from NCI‐H292 cells; (3) prunetin also inhibited the expression of MUC5AC mucin gene induced by EGF or PMA from NCI‐H292 cells. This result suggests that prunetin can regulate the secretion, production and gene expression of mucin, by directly acting on airway epithelial cells. Copyright

Silibinin Regulates Gene Expression, Production and Secretion of Mucin from Cultured Airway Epithelial Cells

We investigated whether silibinin significantly affects gene expression, production and secretion of mucin from cultured airway epithelial cells. Confluent NCI‐H292 cells were pretreated with silibinin for 30 min and then stimulated with epidermal growth factor (EGF), phorbol 12‐myristate 13‐acetate (PMA) or TNF‐α for 24 h. 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). The effect of silibinin on TNF‐α‐induced activation of NF‐κB p65 was also examined. Confluent primary rat tracheal surface epithelial (RTSE) cells were pretreated with adenosine triphosphate (ATP) for 5 min and then treated for 30 min in the presence of silibinin to assess the effect on mucin secretion using ELISA. The results were as follows: (i) silibinin inhibited the expression of the MUC5AC mucin gene induced by EGF, PMA or TNF‐α from NCI‐H292 cells; (ii) silibinin also inhibited the production of MUC5AC mucin protein induced by the same inducers from NCI‐H292 cells; (iii) silibinin inhibited the activation of NF‐κB p65 by TNF‐α in NCI‐H292 cells; (iv) silibinin significantly decreased ATP‐induced mucin secretion from cultured RTSE cells. This result suggests that silibinin can regulate gene expression, production and secretion of mucin by directly acting on airway epithelial cells. Copyright

Phorbol ester or epidermal growth-factor-induced MUC5AC mucin gene expression and production from airway epithelial cells are inhibited by apigenin and wogonin.

In this study, we investigated whether apigenin and wogonin affect MUC5AC mucin production and gene expression induced by phorbol ester (phorbol 12‐myristate 13‐acetate, PMA) or epidermal growth factor (EGF) from human airway epithelial cells. Confluent NCI‐H292 cells were pretreated with each agent for 30 min and then stimulated with PMA or EGF for 24 h, respectively. MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription polymerase chain reaction (RT‐PCR) and enzyme linked immunosorbent assay (ELISA). The results were as follows: (i) apigenin and wogonin were found to inhibit the production of MUC5AC mucin protein induced by PMA or EGF; (ii) both compounds also inhibited the expression of MUC5AC mucin gene induced by PMA or EGF. These results suggest that apigenin and wogonin can inhibit mucin gene expression and production of mucin protein, by directly acting on airway epithelial cells. Copyright

Chondroprotective Effects of Wogonin in Experimental Models of Osteoarthritis in vitro and in vivo.

We evaluated the chondroprotective effects of wogonin by investigating its effects on the gene expression and production of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as on production of MMP-3 in the rat knee. Rabbit articular chondrocytes were cultured in a monolayer, and RT-PCR was used to measure interleukin-1β (IL-1β)-induced expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), and type II collagen. In rabbit articular chondrocytes, the effects of wogonin on IL-1β-induced production and proteolytic activity of MMP-3 were investigated using western blot analysis and casein zymography, respectively. The effect of wogonin on MMP-3 protein production was also examined in vivo. In rabbit articular chondrocytes, wogonin inhibited the expression of MMP-3, MMP-1, MMP-13, and ADAMTS-4, but increased expression of type II collagen. Furthermore, wogonin inhibited the production and proteolytic activity of MMP-3 in vitro, and inhibited production of MMP-3 protein in vivo. These results suggest that wogonin can regulate the gene expression and production of MMP-3, by directly acting on articular chondrocytes.

Effects of Morus alba L. and Natural Products Including Morusin on In Vivo Secretion and In Vitro Production of Airway MUC5AC Mucin

Background It is valuable to find the potential activity of regulating the excessive mucin secretion by the compounds derived from various medicinal plants. We investigated whether aqueous extract of the root bark of Morus alba L. (AMA), kuwanon E, kuwanon G, mulberrofuran G, and morusin significantly affect the secretion and production of airway mucin using in vivo and in vitro experimental models. Methods Effect of AMA was examined on hypersecretion of airway mucin in sulfur dioxide-induced acute bronchitis in rats. Confluent NCI-H292 cells were pretreated with ethanolic extract, kuwanon E, kuwanon G, mulberrofuran G, or morusin for 30 minutes and then stimulated with phorbol 12-myristate 13-acetate (PMA) for 24 hours. The MUC5AC mucin secretion and production were measured by enzyme-linked immunosorbent assay. Results AMA stimulated the secretion of airway mucin in sulfur dioxide-induced bronchitis rat model; aqueous extract, ethanolic extract, kuwanon E, kuwanon G, mulberrofuran G and morusin inhibited the production of MUC5AC mucin induced by PMA from NCI-H292 cells, respectively. Conclusion These results suggest that extract of the root bark and the natural products derived from Morus alba L. can regulate the secretion and production of airway mucin and, at least in part, explains the folk use of extract of Morus alba L. as mucoregulators in diverse inflammatory pulmonary diseases.

Inhibition of TNF-α-Induced MUC5AC Mucin Gene Expression and Production by Wogonin Through the Inactivation of NF-κB Signaling in Airway Epithelial Cells

In this study, we investigated whether wogonin significantly affects MUC5AC mucin gene expression and production in human airway epithelial cells. Confluent NCI‐H292 cells were pretreated with wogonin for 30 min and then stimulated with tumor necrosis factor‐α (TNF‐α) for 24 h or the indicated periods. The MUC5AC mucin gene expression and mucin protein production were measured by RT‐PCR and ELISA, respectively. We found that incubation of NCI‐H292 cells with wogonin significantly inhibited mucin production and down‐regulated MUC5AC gene expression induced by TNF‐α in a dose‐dependent fashion. To elucidate the action mechanism of wogonin, effect of wogonin on TNF‐α‐induced NF‐κB signaling pathway was investigated by western blot analysis. Wogonin inhibited NF‐κB activation induced by TNF‐α. Inhibition of IKK by wogonin led to the suppression of IκB phosphorylation and degradation, p65 nuclear translocation and NF‐κB‐regulated gene expression. This, in turn, led to the down‐regulation of MUC5AC protein production in NCI‐H292 cells. Wogonin also inhibited the gene products involved in cell survival (Bcl‐2) and proliferation (cyclooxygenase‐2). These results suggest that wogonin inhibits the NF‐κB signaling pathway, which may explain its role in the inhibition of MUC5AC mucin gene expression and production. Copyright

Effects of ophiopogonin D and spicatoside A derived from Liriope Tuber on secretion and production of mucin from airway epithelial cells

In the present study, we investigated whether aqueous extract of Liriope Tuber, ophiopogonin D and spicatoside A derived from Liriope Tuber affect basal or phorbol ester (phorbol 12-myristate 13-acetate, PMA)-induced airway mucin production and secretion from airway epithelial cells. Confluent NCI-H292 cells were treated with each agent for 24 h (basal production) or pretreated with each agent for 30 min and then stimulated with PMA for 24 h (PMA-induced production and secretion), respectively. MUC5AC airway mucin production and secretion were measured by ELISA. The results were as follows: (1) aqueous extract of Liriope Tuber stimulated basal mucin production and did not inhibit but increased PMA-induced mucin production; (2) ophiopogonin D and spicatoside A stimulated basal mucin production and did not inhibit but increased PMA-induced mucin production; (3) two compounds increased PMA-induced mucin secretion. These results suggest that ophiopogonin D and spicatoside A can increase mucin production and secretion, by directly acting on airway epithelial cells and, at least in part, explain the traditional use of aqueous extract of Liriope Tuber as expectorants in diverse inflammatory pulmonary diseases.

Resveratrol Inhibits Mucin Gene Expression, Production and Secretion From Airway Epithelial Cells

The study investigated whether resveratrol significantly affects mucin gene expression, production and secretion from airway epithelial cells. Confluent NCI‐H292 cells were pretreated with resveratrol for 30 min and then stimulated with EGF (epidermal growth factor), PMA (phorbol 12‐myristate 13‐acetate) and TNF‐α (tumor necrosis factor‐α) for 24 h, respectively. The MUC5AC gene expression and mucin protein production were measured by RT‐PCR and ELISA. The effect of resveratrol on TNF‐α‐ or PMA‐induced activation of NF‐κB p65 was also examined. Confluent primary rat tracheal surface epithelial (RTSE) cells were pretreated with adenosine triphosphate (ATP) for 5 min and then treated for 30 min in the presence of resveratrol to assess the effect on mucin secretion using ELISA. The results were as follows: (1) resveratrol inhibited the expression of MUC5AC gene induced by EGF or PMA or TNF‐α from NCI‐H292 cells; (2) resveratrol also inhibited the production of MUC5AC mucin protein induced by the same inducers from NCI‐H292 cells; (3) resveratrol inhibited the activation of NF‐κB p65 by TNF‐α or PMA in NCI‐H292 cells; (4) resveratrol significantly decreased ATP‐induced mucin secretion from cultured RTSE cells. This result suggests that resveratrol can regulate mucin gene expression, production and secretion, by directly acting on airway epithelial cells. Copyright

Apigenin Regulates Interleukin-1β-Induced Production of Matrix Metalloproteinase Both in the Knee Joint of Rat and in Primary Cultured Articular Chondrocytes

We examined whether apigenin affects the gene expression, secretion and activity of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as in vivo production of MMP-3 in the knee joint of rat to evaluate the potential chondroprotective effects of apigenin. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcription - polymerase chain reaction (RT-PCR) was used to measure interleukin-1β (IL-1β)-induced expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), and ADAMTS-5. In rabbit articular chondrocytes, the effects of apigenin on IL-1β-induced secretion and proteolytic activity of MMP-3 were investigated using western blot analysis and casein zymography, respectively. The effect of apigenin on MMP-3 protein production was also examined in vivo. In rabbit articular chondrocytes, apigenin inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5. Furthermore, apigenin inhibited the secretion and proteolytic activity of MMP-3 in vitro, and inhibited production of MMP-3 protein in vivo. These results suggest that apigenin can regulate the gene expression, secretion, and activity of MMP-3, by directly acting on articular chondrocytes.