• The Journal of Korean Medicine
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• v.29 no.4
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• pp.123-132
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• 2008

Objective: In this study, the author tried to investigate whether revised Seonbangpaedok-tang (SPT) and Sigyeongcheongpye-tang (SCT) significantly effects both PMA-induced mucin production and MUC5AC gene expression from airway epithelial cells. Objective: In this study, the author tried to investigate whether revised Seonbangpaedok-tang (SPT) and Sigyeongcheongpye-tang (SCT) significantly affects both PMA-induced mucin production and MUC5AC gene expression from airway epithelial cells. Materials and Methods: Confluent NCI-H292 cells were pretreated for 30 min in the presence of SPT and SCT and treated with PMA (10 ng/$m{\ell}$), to assess the effects of the agents on PMA-induced mucin production by enzyme-linked immunosorbent assay (ELISA). Also, the effects of the agents on PMA-induced MUC5AC gene expression from the same cells were investigated. Possible cytotoxicities of the agent were assessed by measuring the rate of survival and proliferation of NCI-H292 cells after treatment of agents during 48 hrs. Results: (1) SPT and SCT did not show significant cytotoxicity to NCI-H292 cells; (2) SPT significantly inhibitedthe expression levels of PMA-induced MUC5AC gene in NCI-H292 cells. SCT slightly decreased the expression levels of PMA-induced MUC5AC gene; (3) SPT significantly decreased PMA-induced mucin production from NCI-H292 cells. However, SCT did not affect mucin production. Conclusion: Theseresults suggest that SPT can not only affect the production of mucin but also the expression of the mucin gene and this explains the traditional use of SPT in oriental medicine. The effects of SPT and SCT with their components should be further investigated using animal experimental models that reflect pathophysiology of airway diseases via ongoing studies.

• The Journal of Internal Korean Medicine
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• v.29 no.2
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• pp.318-333
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• 2008

Objectives : In this study, the author tried to examine whether Cheogjogupye-tang (淸燥救肺湯, CGPT) and Yieum-jeon (理陰煎, YEJ) significantly affect in vitro and in vivo mucin secretion, MUC5AC gene expression in airway epithelial cells and contractility of isolated tracheal smooth muscle of rabbit. Materials and Methods : For in vitro experiment, confluent hamster tracheal surface epithelial (HTSE) cells were chased for 30 minutes in the presence of CGPT and YEJ to assess the effects of the agents on mucin secretion by enzyme-linked immunosorbent assay (ELISA), with removal of oriental herbal medicine extract from each agent-treated sample by centrifuge microfilter. Also, the effects of the agents on TNF-alpha or EGF-induced MUC5AC gene expression in human airway epithelial cells (NCI-H292) were investigated. Possible cytotoxicities of the agent were assessed by examining both LDH release from HTSE cells and the rate of survival and proliferation of NCI-H292 cells. For in vivo experiment, hypersecretion of airway mucin and goblet cell hyperplasia was induced by exposure of rats to $SO_2$ over 3 weeks. Effects of CGPT and YEJ orally administered for 1 week on in vivo mucin secretion from tracheal goblet cells of rats and hyperplasia of goblet cells were assessed using ELISA and histological analysis after staining the epithelial tissue with alcian blue, respectively. Also, the effects of CGPT and YEJ on contractility of isolated tracheal smooth muscle were investigated. Results : (1) CGPT significantly inhibited in vitro mucin secretion from cultured HTSE cells. However, YEJ did not affect in vitro mucin secretion; (2) CGPT and YEJ did not affect hypersecretion of in vivo mucin and hyperplasia of tracheal goblet cells; (3) CGPT and YEJ slightly increased the expression levels of TNF-alpha or EGF-induced MUC5AC gene in NCI-H292 cells; (4) CGPT and YEJ inhibited acetylcholine-induced contraction of isolated tracheal smooth muscle of rabbit; (5) CGPT and YEJ did not affect LDH release from HTSE cells and the survival and proliferation of NCI-H292 cells. Conclusion : The results from the present study suggest that CGPT and YEJ mainly affect the expression of mucin gene rather than secretion of mucin and do not show remarkable cytotoxicity to respiratory epithelial cells.

• Biomolecules & Therapeutics
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• v.29 no.3
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• pp.303-310
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• 2021

In the present study, kaempferol, a flavonoidal natural compound found in Polygonati Rhizoma, was investigated for its potential effect on the gene expression and production of airway MUC5AC mucin. A human respiratory epithelial NCI-H292 cells was pretreated with kaempferol for 30 min and stimulated with epidermal growth factor (EGF) or phorbol 12-myristate 13-acetate (PMA), for the following 24 h. The effect on PMA-induced nuclear factor kappa B (NF-κB) signaling pathway or EGF-induced mitogen-activated protein kinase (MAPK) signaling pathway was investigated. Kaempferol suppressed the production and gene expression of MUC5AC mucins, induced by PMA through the inhibition of degradation of inhibitory kappa Bα (IκBα), and NF-κB p65 nuclear translocation. Also, kaempferol inhibited EGF-induced gene expression and production of MUC5AC mucin through regulating the phosphorylation of EGFR, phosphorylation of p38 MAPK and extracellular signal-regulated kinase (ERK) 1/2 (p44/42), and the nuclear expression of specificity protein-1 (Sp1). These results suggest kaempferol regulates the gene expression and production of mucin through regulation of NF-κB and MAPK signaling pathways, in human airway epithelial cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.1
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• pp.98-103
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• 2007

In the present study, the author intended to investigate whether Gamiyukgunja-tang (Jiaweiliujunzi-tang, GYGT) significantly affect both mucin release from and MUC5AC gene expression in cultured hamster tracheal surface epithelial (HTSE) cells. Confluent HTSE cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of GYGT to assess the effect on 3H-mucin release. Possible cytotoxicity of the agent was assessed by measuring lactate dehydrogenase (LDH) release. Total elution profiles of control spent media and treatment sample through Sepharose CL-4B column were analysed and effect of GYGT on MUC5AC gene expression in cultured HTSE cells were investigated. GYGT did not affect mucin release from cultured HTSE cells. GYGT did not show significant cytotoxicity. GYGT also did not affect the secretion of the other releasable glycoproteins with less molecular weight than mucin. GYGT increased the expression level of MUC5AC gene. We suggest that the effect of GYGT with their components should be further investigated through ongoing research.

• Biomolecules & Therapeutics
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• v.13 no.4
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• pp.246-250
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• 2005

In this study, we tried to investigate whether diclofenac, acetaminophen, nimesulide, acetylsalicylic acid and tumor necrosis factor-alpha (TNF-alpha) significantly affect mucin release from cultured airway goblet cells. Confluent primary hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with $^3H$-glucosamine for 24 hr and chased for 30 min or 24 hr in the presence of each agent to assess the effects on $^3H$-mucin release. The results were as follows: (1) TNF-alpha significantly increased mucin release from cultured HTSE cells during 24 hr of treatment period; (2) However, diclofenac, acetaminophen, nimesulide and acetylsalicylic acid did not affect mucin release, during 30 min of treatment period. Basically, this finding suggests that non-steroidal antiinflammatory drugs (NSAIDs) might not function as a mucoregulator in various inflammatory respiratory diseases showing mucus hypersecretion, although further studies are needed.

• The Journal of Internal Korean Medicine
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• v.25 no.2
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• pp.238-244
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• 2004

Objective : This study is intended to investigate whether the two oriental medical prescriptions, CheongGeumGangHwa-tang(CGGH) and GwaRuJiSil-tang(GRJS), significantly affect mucin release from cultured hamster tracheal surface epithelial(HTSE) cells. Materials and Methods : Confluent HTSE cells were metabolically radio labeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of CGGH or GRJS to assess the effect of each agent on 3H-mucin release. Possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase(LDH) release. Also, the effects of CGGH and GRJS on contractility of isolated tracheal smooth muscle were investigated. Results : (1) CGGH and GRJS significantly increased mucin release from cultured HTSE cells, without cytotoxicity : (2) CGGH and GRJS did not affect contractility of isolated tracheal smooth muscle. Conclusions : These results suggest that the effects of CGGH and GRJS should be further investigated, and that it would be gainful to invesigate, from among oriental medical prescriptions, what novel agents have these mild expectorant effects on mucin secretion from airway goblet cells.

• Biomolecules & Therapeutics
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• v.9 no.4
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• pp.263-269
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• 2001

In this study, we tried to investigate whether poly-L-arginine (PLA) (MW 10,800) significantly affect mucin release from cultured hamster airway goblet cells and the mucosubstances of hypersecretory air-way goblet cells of rats. Confluent primary hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with $^3$H-glucosamine for 24 hr and chased for 30 min in the presence of varying concentrations of PLA to assess the effects on $^3$H-mucin release. Possible cytotoxicities of PLA were assessed by measuring both Lactate Dehydrogenate (LDH) release and by checking the possible changes on the morphology of HTSE cells during treatment. For in vivo experiment, hyperplasia of rat airway goblet cells and increase in intraepithelial mucosubstances were induced by exposing rats to SO$_2$ for 3 weeks and varying concentrations of PLA were administered inhalationally to assess the effects on the mucosubstances of airway goblet cells of rats. The results were as follows : (1) PLA significantly inhibited mucin release from cultured HTSE cells in a dose-dependent manner; (2) there was no significant release of LDH and no significant change on the morphology of cultured HTSE cells during treatment; (3) PLA also affected the intraepithelial mucosubstances of hypersecretory rats and restored them to the levels of control animals. We conclude that PLA inhibit mucin release from airway goblet cells without significant cytotoxicity and possibly normalize the hypersecretion of airway mucosubstances in vivo. This finding suggests that PLA might function as an airway mucoregulative agent.

• The Journal of Internal Korean Medicine
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• v.26 no.1
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• pp.221-228
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• 2005

The intent of this study is to investigate whether two oriental medical prescriptions named haengsotang(HST) and gami-palmihwan(GPMH) significantly effect mucin release from cultured hamster tracheal surface epithelial(HTSE) cells, Confluent HTSE cells were metabolically radiolabeled with $^3H-glucosamine$ for 24 hrs and chased for 30 min in the presence of HST or GPMH to assess the effect of each agent on $^3H-mucin$ release. Possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase(LDH) release. Also, the effects of HST and GPMH on contractility of isolated tracheal smooth muscle were investigated. The results are consistant with the following assertions: (1) HST significantly inhibited mucin release from cultured HTSE cells, without cytotoxicity; (2) GPMH did not effect mucin release without cytotoxicity; (3) HST and GPMH did not effect contractility of isolated tracheal smooth muscle. These results suggest a need for further investigation of HST and its components, for its potential in oriental medicine prescriptions and novel agents that effectively regulate (inhibit) mucin secretion from airway goblet cells.

• Biomolecules & Therapeutics
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• v.22 no.6
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• pp.525-531
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• 2014

In the present study, we investigated whether apigenin significantly affects tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-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-${\alpha}$ 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-${\alpha}$ in NCI-H292 cells. To elucidate the action mechanism of apigenin, effect of apigenin on TNF-${\alpha}$-induced nuclear factor kappa B (NF-${\kappa}B$) signaling pathway was also investigated by western blot analysis. Apigenin inhibited NF-${\kappa}B$ activation induced by TNF-${\alpha}$. Inhibition of inhibitory kappa B kinase (IKK) by apigenin led to the suppression of inhibitory kappa B alpha ($I{\kappa}B{\alpha}$) 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-${\kappa}B$ signaling pathway in airway epithelial cells.

• The Journal of Internal Korean Medicine
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• v.30 no.4
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• pp.685-695
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• 2009

Objectives : In this study, the author tried to investigate whether Geonpye-tang(GPT) significantly affects PMA-, EGF- or TNF-alpha-induced MUC5AC mucin production and gene expression from human airway epithelial cells. Materials and Methods : Effects of the agent on PMA-, EGF- or TNF-alpha-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of GPT and treated with PMA (10ng/ml) or EGF (25ng/ml) or TNF-alpha (0.2nM), to assess both effect of the agent on PMA- or EGF- or TNF-alpha-induced MUC5AC mucin production by enzyme-linked immunosorbent assay (ELISA) and gene expression by reverse transcription-polymerase chain reaction (RT-PCR). Possible cytotoxicity of the agent was assessed by examining the rate of survival and proliferation of NCI-H292 cells after treatment with the agent over 72 hrs (SRB assay). Results : (1) GPT significantly inhibited PMA-induced and EGF-induced MUC5AC mucin production from NCI-H292 cells. However, GPT did not affect TNF-alpha-induced MUC5AC mucin production. (2) GPT significantly inhibited the expression levels of PMA-, EGF- or TNF-alpha-induced MUC5AC genes in NCI-H292 cells (3) GPT did not show significant cytotoxicity to NCI-H292 cells. Conclusion : This result suggests that GPT can affect the production and gene expression of respiratory mucin observed in diverse respiratory diseases accompanied by mucus hypersecretion. This can explain the traditional use of GPT in oriental medicine. Effects of GPT with their components should be further investigated using animal experimental models that reflect pathophysiology of airway diseases through future studies.