• 동의생리병리학회지
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• 제21권1호
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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.

• 대한한방소아과학회지
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• 제20권1호
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• pp.109-135
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• 2006

Objective : In the present study, the author tried to investigate whether six oriental medical prescriptions named gamisingitang (SGT), gamijungtang (IJT), gamicheongpyetang (CPT), galhwengchihyosan (CHS), chwiyeontong (CYT), sigyoungcheongpyetang (SCPT) significantly affect mucin release from cultured hamster tracheal surface epithelial (HTSE) cells. Methode : Confluent HTSE cells were inetabolically radiolabeled with $^{3}H-glucosamine$ for 24 hrs and chased for 30 min in the presence of drugs aforementioned, respectively, to assess the effect of each drug on $^{3}H-mucin$ release. Possible cytotoxicities of effective drugs were assessed by measuring lactate dehydrogenase(LDH) release. Additionally, total elution profiles of control spent media and treatment sample (CPT, CHS, SCPT and CYT) through Sepharose CL-4B column were analysed and effect of CPT, CHS and CYT on MUC5AC mRNA expression in cultured HTSE cells were invsetigated. Results : (1) SGT and IJT did not affect mucin release without cytotoxicity; (2) CPT, SCPT and CHS significantly stimulated mucin release from cultured HTSE cells, with significant cytotoxicity; (4) CPT, CHS, SCPT and CYT chiefly affected the 'mucin' release and did not affect significantly the release of the releasable glycoproteins with less molecular weight than mucin. This result suggests that the four herbal prescriptions specifically affect the release of mucin ; (5) CTP and CHS did not significantly affect the expression levels of MUC 5AC mRNA, however, CYT significantly inhibit the expression levels of MUC 5AC mRNA. Conclusion : CYT can decrease the synthesis of mucin at gene level in cultured HTSE cells.

• 동의생리병리학회지
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• 제18권3호
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• pp.734-739
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• 2004

In the present study, the author intended to investigate whether two oriental medical prescriptions named socheongryong-tang(SCRT) and Kamichihyo-san(KCHS) 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 SCRT or KCHS 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 SCRT and KCHS on contractility of isolated tracheal smooth muscle were investigated. The results were as follows: (1) SCRT significantly inhibited mucin release from cultured HTSE cells, without cytotoxicity; (2) KCHS significantly increased mucin release without cytotoxicity; (3) SCRT and KCHS did not affect contractility of isolated tracheal smooth muscle. We suggest that the effects of SCRT and its components should be further investigated and it is of great value to find, from oriental medical prescriptions, novel agents which have the possible inhibitory effects on mucin release from the viewpoint of management of hypersecretion of airway mucus.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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• pp.198-198
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• 1998

Muc1 mucin is found in a variety of epithelial tissue and is overexpressed in several epithelial cancer. Recently it is alsol reported that primary Hamster tracheal surface epithelial(HTSE) cells express Muc1 protein and cDNA encoding HTSE muc1 protein has been cloned. Although numerous monoclonal antibodies (mAbs) to human muncins, particularly Muc1 have been produced, no such antibodies to murine Muc1 have been described. We now describe monoclonal antibody, called mAb M1CT, produced to C-terminal region of HTSE Muc1 protein by immunising mice with a glutathion-s-transferase linked fusion protein. In this study, using this antibody(mAb M1CT) we investigated the effect of RA on the expression of Muc1 in HTSE cells. Retinoic acid(RA) plays an essential role in maintaining normal differentiation of tracheal epithelial cells. With RA-deficiency tracheocytes undergo squamous metaplasia, an abnormal differentiation that can be reversed by RA. We had primary culture of HTSE cells under different concentrations of RA. Culture was maintained until the direction of differentiation was determined. Then Western blot analysis with mAb M1CT was performed with the cell lysates from the culture. The expression of Muc1 protein was decreased in dose-dependent manner as the concentration of retinoic acid was decreased. Our result indicates that the expression of Muc1 protein is coordinately regulated with airway mucous cell differentiation by RA pathway. And the antibody, mAb M1CT, produced in this study should provide useful tool to study the expression of Muc1 mucin in differentiation process or disease.

• 대한한방내과학회지
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• 제28권4호
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• pp.797-807
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• 2007

Objectives : The purpose of this study was to investigate whether Mahwangyunpye-tang(MYT) significantly affects mucin secretion from airway epithelial cells. Methods : Confluent hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of MYT to assess the effect of the agent on 3H-mucin secretion. Total elution profiles of control spent media and treatment sample through Sepharose CL-4B column were analyzed. Effect of MYT on contractility of isolated tracheal smooth muscle was investigated; also investigated was effect of the agent on MUC5AC gene expression in cultured NCI-H292 cells. Possible cytotoxicities of the agent were assessed both by measuring lactate dehydrogenase (LDH) release from HTSE cells and examining the rate of survival and proliferation of NCI-H292 cells. Results : MYT significantly increased mucin secretion from cultured HTSE cells, without significant cytotoxicity. MYT chiefly affected the 'mucin' secretion. MYT inhibited Acetylcholine-induced contraction of isolated tracheal smooth muscle. MYT did not significantly affect the expression levels of MUC 5AC gene in cultured NCI-H292 cells. Conclusions : Based on the above results, it is suggested that MYT increased mucin secretion from cultured HTSE cells without significant cytotoxicity and inhibited contraction of isolated tracheal smooth muscle.

• 대한한의학회지
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• 제29권3호
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• pp.76-87
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• 2008

Objectives: In this study, the author investigated whether Haengso-tang (HST) and Chwiyeon-tang (CHT) affect both in vitro mucin secretion and MUC5AC gene expression in airway epithelial cells and in vivo mucin secretion from animal model for airway mucus hypersecretion. Materials and Methods: Confluent HTSE cells (non-labeled) were chased for 30 min in the presence of HST and CHT 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- or EGF-induced MUC5AC gene expression in human airway epithelial cells (NCI-H292) were investigated. The author also induced hypersecretion of airway mucus by exposure of rats to SO2 for 3 weeks. Effects of orally-administered HST and CHT during 1 week on in vivo mucin secretion from tracheal goblet cells of rats were assessed using ELISA. Results: (1) HST significantly decreased in vitro mucin secretion from cultured HTSE cells. However, CHT did not affect in vitro mucin secretion from HTSE cells; (2) CHT significantly inhibited the expression levels of EGF- or TNF-alpha-induced MUC5AC gene in NCI-H292 cells. However, HST did not affect the expression levels of EGF- or TNF-alpha-induced MUC5AC gene in NCI-H292 cells; (3) CHT significantly inhibited hypersecretion of in vivo mucin. However, HST did not affect hypersecretion of in vivo mucin. Conclusion: These results suggest that CHT can not only affect the secretion of mucin but also the expression of the mucin gene and could be helpful for treating pulmonary disease caused by secretion of mucin.

• 대한한방내과학회지
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• 제27권2호
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• pp.295-304
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• 2006

Objectives : This study was done to investigate whether two oriental prescriptions, saganmahwang-tang (SMT) and prescription C (P-C) significantly affect mucin release from cultured hamster tracheal surface epithelial (HTSE) cells. Methods : Cofluent HTSE cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of SMT or P-C 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 SMT and P-C on contractility of isolated tracheal smooth muscle were investigated. Results : SMT significantly inhibited mucin release from cultured HTSE cells, without cytotoxicity. P-C significantly increased mucin release from cultured HTSE cells, with significant cytotoxicity. SMT inhibited Ach-induced contraction of isolated tracheal smooth muscle. P-C did not affect Ach-induced contraction of isolated tracheal smooth muscle. Conclusion : Results su99est that SMT and P-C have regulating effects on mucin secretion from airway goblet cells. Further investigation is needed, because of the value in finding novel agents to this purpose, and these oriental medical prescription have potential for such a role.

• 대한한방소아과학회지
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• 제20권1호
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• pp.93-107
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• 2006

Objective : To investigate the effectiveness of two oriental medical prescriptions named Sigoungcheongpyetang(SCPT) and Tonggyutanggamibang(TGT) for mucin release from cultured hamster tracheal surface epithelial(HTSE) cells. Method : Confluent HTSE cells were metabolically radiolabeled with $^{3}H-glucosamine$ for 24hrs and chased for 30 min in the presence of SCPT or TGT to assess the effect of each agent $^{3}H-mucin$ release. Possible cytotoxicities of each agent were assessed by measuring lactate dehy drogenase(LDH) release. Also, the effects of SCPT and TGT on contrectility of isolated tracheal smooth muscle were investigated. Results : (1) SCPT and TGT significantly increased mucin release from cultured HTSE cells, with significantly cytotoxicity ; SCPT did not affect contractility of isolated tracheal smooth muscle and TGT inhibited Ach-induced contraction of isolated tracheal smooth muscle. Conclusion : We suggest that the effects of SCPT and TGT with their components should be further investigated and it is of great value to find, from oriental medical prescriptions, novel agents which might regulate mucin secretion from airway goblet cells.

• 동의생리병리학회지
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• 제20권1호
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• pp.156-162
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• 2006

In the present study, the author intended to investigate whether two oriental medical prescriptions named GSGT and GCPT 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 presen+ce of GSGT or GCPT to assess the effect of each agent on 3H-mucin release. Possible cytotoxicities of each agent were assessed dy measuring lactate dehydrogenase(LDH) release. Also, the effects of GSGT and GCPT on contractility of isolated tracheal smooth muscle were investigated. (1) GSGT did not affect mucin release without cytotoxicity ; (2) GCPT significantly stimulated mucin release from cultured HTSE cells, with significant cytotoxicity ; (3) GSGT and GCPT did not affect contractility of isolated tracheal smooth muscle. We suggest that the effects of GCPT and its components should be further investigated and it is of great value to find, from oriental medical prescriptions, novel agents which have potent expectorant effects on mucin secretion from airway goblet cells.

• 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.