• Biomolecules & Therapeutics
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• 제14권3호
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• pp.178-182
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• 2006

In this study, we investigated whether daidzein, puerarin, genistein and (+)-ar-tumerone affect mucin secretion from cultured airway epithelial cells and compared with the inhibitory action of poly-L-lysine (PLL) and the stimulatory action of adenosine triphosphate (ATP) on mucin secretion. Confluent primary hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled using $^3H$-glucosamine for 24 h and chased for 30 min in the presence of varying concentrations of each agent to assess the effects on $^3H$-mucin secretion. The results were as follows: daidzein, puerarin, genistein and (+)-ar-tumerone did not affect mucin secretion at the highest concentrations $(10^{-3}M)$, during 30 min of treatment period. Basically, this finding suggests that daidzein, puerarin and genistein - 3 components derived from Puerariae Radix - and (+)-ar-tumerone derived from Curcumae Rhizoma might not function as a mucoregulator in various inflammatory respiratory diseases showing mucus hypersecretion, although further studies are needed.

• 동의생리병리학회지
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• 제20권1호
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• pp.69-75
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• 2006

In the present study, the author tried to investigate whether four oriental medical prescriptions named, jawan-chihyosan (CHS), gwaru-jisiltang (GJT), and several single compounds, kaempferol, coumarin, betaine and ursolic acid significantly affect 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 CHS, GJT, kaempferol, coumarin, betaine and ursolic acid, respectively, to assess the effect of each agent on 3H-mucin release. Possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase(LDH) release. Additionally, total elution profiles of control spent media and treatment sample (CHS and GJT) through Sepharose CL-4B column were analysed and effect of CHS and GJT on MUC5AC mRNA expression in cultured HTSE cells were investigated. The results were as follows : (1) CHS and GJT significantly stimulated mucin release from cultured HTSE cells, with significant cytotoxicity , (2) CHS and GJT chiefly stimulated the 'mucin' release and did not affect significantly the release of the other releasable glycoproteins with less molecular weight than mucin. This result suggests that the three herbal prescriptions specifically stimulate the release of mucin ; (3) CHS and GJT significantly increased the expression levels of MUC 5AC mRNA. This result suggests that the three herbal prescriptions can affect the synthesis of mucin at gene level in cultured HTSE cells ; (4) Kaempferol and coumarin did not affect mucin release, however, betaine and ursolic acid stimulated mucin release. All the agents did not show significant cytotoxicity. We suggest that the effects of CHS and GJT, betaine and ursolic acid should be further investigated and it is of great value to find, from oriental medical prescriptions, novel agents which have the effective expectorant or mucoregulative effect on mucin secretion from airway goblet cells.

• Journal of Microbiology and Biotechnology
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• 제27권1호
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• pp.49-56
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• 2017

In the human airway, mucus exists to protect the respiratory system as a primary barrier of the innate immune system. However, hyperexpressed mucus limits airflow, resulting in a decrease of lung function. Among more than 20 mucin family members, MUC5AC and MUC5B are major glycoproteins in human airway mucus. The epidermal growth factor receptor (EGFR) signaling pathway is one of the mechanisms of these mucins expression and specificity protein-1 (Sp1) transcription factor is the downstream signal of this pathway, playing pivotal roles in mucin expression. Even though there are some drugs for treating mucus hypersecretion, no drug has proven effects on humans. We found that the flavonoid tilianin regulated MUC5AC expression and also inhibited Sp1 phosphorylation. In this study, we investigated how tilianin would modulate EGFR signaling and regulate mucin production. In conclusion, tilianin inhibited MUC5AC expression mediated via modulating the EGFR-MEK-ERK-Sp1 signaling pathway in NCI-H292 human airway epithelial cells. This study may provide the basis for the novel treatment of mucus hypersecretion.

• Journal of Ginseng Research
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• 제46권6호
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• pp.801-808
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• 2022

Background: Diesel exhaust particle (DEP) is a harmful kind of particulate matter known to exacerbate pre-existing respiratory diseases. Although their adverse effects on airway pathologies have been widely studied, the mechanistic analysis of signaling pathways and potential targets in reducing DEP-induced mucin secretion and pro-inflammatory cytokine production remain elusive. We, for the first time, investigated the effects of Korean Red Ginseng (KRG) extracts on mucin overproduction and airway inflammation induced by DEP. Methods: The effects of KRG and saponin on DEP-induced expression of MUC5AC and interleukin (IL)-6/8 were examined by real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) in human airway epithelial NCI-H292 cells. We conducted Western blotting analysis to analyze the associated signaling pathways. To evaluate the effects of saponin treatment on DEP-induced MUC5AC expression and inflammatory cell infiltrations in ovalbumin (OVA)-sensitized mice, immunohistochemical (IHC) staining and real-time PCR were implemented. Results: The KRG extracts markedly attenuated DEP-induced MUC5AC expression in vitro by inhibiting the TLR4/TRIF/NF-𝛋B pathway. Furthermore, KRG and saponin inhibited DEP-induced pro-inflammatory cytokine IL-6/8 production. The in vivo study revealed that saponin blocked DEP-induced inflammation, mucin production and MUC5AC expression. Conclusion: Our study revealed that KRG extracts have inhibitory effects on DEP-induced expression of MUC5AC and the production of pro-inflammatory cytokines. This finding provides novel insights into the mechanism by which saponin alleviates diesel-susceptible airway inflammation, elucidating its potential as a phytotherapeutic agent for inflammatory pathologies of airway.

• 대한한방소아과학회지
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• 제19권1호
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• pp.11-23
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• 2005

Objectives : In the present study, the authors intended to investigate whether two oriental medical prescriptions named chwiyeon-tang and chihyosan-gamibang significantly affect much release from cultured hamster tracheal surface epithelial HTSE cells. Methods : Confluent HTSE cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of chwiyeon-tang or chihyosan-gamibang 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 effect of chwiyeon-tang and chihyosan-gamibang on contractility of isolated tracheal smooth muscle were investigated. Results : (1) Chwiyeon-tang significantly inhibited mucin release from cultured HTSE cells, with significant cytotoxicity ; (2) Chihyosan-gamibang significantly stimulated mucin release from cultured HTSE cells, with minute cytotoxicity ; (3) Chwiyeon-tang and Chihyosan-gamibang did not affect contractility of isolated tracheal smooth muscle. Conclusions : We suggest that the effects of Chwiyeon-tang and Chihyosan-gamibang with their components should be further investigated and it is of great value to find, from oriental medical prescriptions, noel agents which might regulate mucin secretion from airway goblet cells.

• Biomolecules & Therapeutics
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• 제28권5호
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• pp.431-436
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• 2020

In this study, diclofenac, a non-steroidal anti-inflammatory drug, was investigated for its potential effect on the gene expression and production of airway MUC5AC mucin. The human respiratory epithelial NCI-H292 cells were pretreated with diclofenac for 30 min and stimulated with phorbol 12-myristate 13-acetate (PMA), for the following 24 h. The effect of diclofenac on PMA-induced nuclear factor kappa B (NF-kB) signaling pathway was also investigated. Diclofenac suppressed the production and gene expression of MUC5AC mucins, induced by PMA through the inhibition of degradation of inhibitory kappa Bα (IkBα) and NF-kB p65 nuclear translocation. These results suggest diclofenac regulates the gene expression and production of mucin through regulation of NF-kB signaling pathway, in human airway epithelial cells.

• 대한한방소아과학회지
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• 제27권1호
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• pp.69-81
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• 2013

Objectives In this study, effects of Macmundongtang (MMT) on ATP or TNF-${\alpha}$ or PMA or EGF induced MUC5AC mucin production and gene expression from human airway epithelial cells and the increase in airway epithelial mucosubstances of rats were investigated. Materials and Methods Confluent NCI-H292 cells were pretreated for 30min in the presence of MMT and treated with ATP ($200{\mu}M$) or PMA (10 ng/ml) or EGF (25 ng/ml) or TNF-${\alpha}$ (0.2 nM) for 24hrs, to assess the effect of MMT both on ATP- or PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production using enzyme-linked immunosorbent assay (ELISA) and on gene expression by the same inducers using reverse transcription-polymerase chain reaction (RT-PCR). At the same time, hypersecretion of airway mucus was induced by exposure of rats to SO2 during 3 weeks. Effect of orally-administered MMT during 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats was assesed using histopathological analysis after staining the epithelial tissue with PAS-alcian blue. Possible cytotoxicity of MMT was assessed by investigating the potential damage of kidney and liver functions by measuring serum GOT/GPT activities and serum BUN concentration of rats and the body weight gain during experiment, after administering MMT orally. Results (1) MMT did not only inhibit but also increased MUC5AC mucin productions and expression levels of MUC5AC gene from NCI-H292 cells. (2) MMT did not decrease the amount of intraepithelial mucosubstances of trachea of rats. (3) MMT did not show renal and hepatic toxicities and did not affect body weight gain of rats during experiment. Conclusions The result from the present study suggests that MMT might normalize the production and gene expression of airway mucin observed in various respiratory diseases accompanied by yin-deficiency, without in vivo toxicity to liver and kidney functions after oral administration.

• 치위생과학회지
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• 제2권1호
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• pp.25-29
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• 2002

본 연구에서는 분자량 78,000과 9,600의 poly-L-lysine(PLL)이 일차배양된 햄스터의 기관표면 상피세포로부터의 뮤신유리에 어떠한 영향을 미치는지를 검증하고자 하였다. 완전히 다 자란 배양세포에 $^3H$-glucosamine을 함유하는 완전 배양액을 첨가하고 24시간 배양함으로써 배양세포 중의 뮤신에 대사적 방사선 표지를 완결한 후 다양한 농도의 PLL을 30분간 처리하고, 유리되는 방사성 뮤신의 함량을 측정하였다. PLL에 의한 세포독성 발현여부를 검증하기 위하여 PLL 처리 후 배양세포로부터 유리되어 배양액 중에 존재하는 Lactate Dehydrogenase(LDH)의 활성을 측정하였다. 실험 결과, PLL은 분자량 78,000 및 9,600의 두 물질 공히 용량 의존적으로 뮤신유리를 억제하였다. 그러나 세포독성의 지표인 LDH 유리에 대한 영향은 PLL 9,600의 경우에는 유의성이 없었으나, PLL 78,000의 경우에는 현저한 유리 증가를 보였다. 이러한 결과는 PLL의 경우, 분자량이 10,000 이하의 범위에서 세포독성을 발현하지 않으면서도 뮤신유리를 특이적으로 억제할 가능성을 제시하고 있으며, 동시에 PLL이 기도점액 과다분비 현상을 연구함에 있어서 유용한 실험수단으로 이용될 가능성도 제시하고 있는 것이다.

• Tuberculosis and Respiratory Diseases
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• 제76권3호
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• pp.120-126
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• 2014

Background: We investigated whether wogonin and apigenin significantly affect the epidermal growth factor receptor (EGFR) signaling pathway involved in MUC5AC mucin gene expression, and production from cultured airway epithelial cells; this was based on our previous report that apigenin and wogonin suppressed MUC5AC mucin gene expression and production from human airway epithelial cells. Methods: Confluent NCI-H292 cells were pretreated with wogonin or apigenin for 15 minutes or 24 hours and then stimulated with epidermal growth factor (EGF) for 24 hours or the indicated periods. Results: We found that incubation of NCI-H292 cells with wogonin or apigenin inhibited the phosphorylation of EGFR. The downstream signals of EGFR such as phosphorylation of MEK1/2 and ERK1/2 were also inhibited by wogonin or apigenin. Conclusion: The results suggest that wogonin and apigenin inhibits EGFR signaling pathway, which may explain how they inhibit MUC5AC mucin gene expression and production induced by EGF.

• Journal of Ginseng Research
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• 제47권1호
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• pp.97-105
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• 2023

Background: Hyperactivated airway mucosa cells overproduce mucin and cause severe breathing complications. Here, we aimed to identify the effects of saponins derived from Panax ginseng on inflammation and mucin overproduction. Methods: NCI-H292 cells were pre-incubated with 16 saponins derived from P. ginseng, and mucin overproduction was induced by treatment with phorbol 12-myristate 13-acetate (PMA). Mucin protein MUC5AC was quantified by enzyme-linked immunosorbent assay, and mRNA levels were analyzed using quantitative polymerase chain reaction (qPCR). Moreover, we performed a transcriptome analysis of PMA-treated NCI-H292 cells in the absence or presence of Rg5, and differential gene expression was confirmed using qPCR. Phosphorylation levels of signaling molecules, and the abundance of lipid droplets, were measured by western blotting, flow cytometry, and confocal microscopy. Results: Ginsenoside Rg5 effectively reduced MUC5AC secretion and decreased MUC5AC mRNA levels. A systematic functional network analysis revealed that Rg5 upregulated cholesterol and glycerolipid metabolism, resulting in the production of lipid droplets to clear reactive oxygen species (ROS), and modulated the mitogen-activated protein kinase and nuclear factor (NF)-kB signaling pathways to regulate inflammatory responses. Rg5 induced the accumulation of lipid droplets and decreased cellular ROS levels, and N-acetyl-ⳑ-cysteine, a ROS inhibitor, reduced MUC5AC secretion via Rg5. Furthermore, Rg5 hampered the phosphorylation of extracellular signal-regulated kinase and p38 proteins, affecting the NF-kB signaling pathway and pro-inflammatory responses. Conclusion: Rg5 alleviated inflammatory responses by reducing mucin secretion and promoting lipid droplet-mediated ROS clearance. Therefore, Rg5 may have potential as a therapeutic agent to alleviate respiratory disorders caused by hyperactivation of mucosa cells.