• Biomolecules & Therapeutics
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• v.6 no.1
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• pp.14-19
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• 1998

In the present study, we intended to investigate whether cationic polyamines including poly-L-Iysine (PLL) and poly-L-arginine (PLA) induce cytotoxicities to cultured hamster tracheal surface epithelial (HTSE) cells. Confluent HTSE cells were chased for 30 min in the presence of PLL or PLA of different molecular weights. Possible cytotoxicities of PLL or PLA were assessed by measuring both Lactate Dehy- drogenase (LDH) release during treatment and the number of floating cells after treatment and by checking the possible changes on the morphology of HTSE cells during treatment. The results were as follows: in the case of treatment of PLL or rLA of which molecular weight is about 78,000 and 92,000, respectively, (1) there was significant release of LDH during treatment, (2) the number of floating cells were significantly increased after treatment and (3) there were significant changes on the morphology of cultured HTSE cells. However, in the case of PLL or PLA of which molecular weight is under 10,000 (about 9,600 and 8,900, respectively), no significant signs of cytotoxicities mentioned above were detected. We found that cationic polyamines might be non-toxic under specific range of molecular weights and suggest that the cytotoxicity of cationic polyamine might depend on the molecular sizes of each cationic polyamine.

• Biomolecules & Therapeutics
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• v.10 no.3
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• pp.156-164
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• 2002

In the present study, we tried to investigate whether polymerized basic amino acid e.g. poly-L-lysine (PLL) which has the degree of polymerization under 50mer significantly affects the physiological and stimulated mucin release from cultured hamster tracheal surface epithelial cells. 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 either PLLs or adenosine triphosphate (ATP) and PLL to assess the effects on basic or ATP-stimulated $^3{H}$-mucin release. Possible cytotoxicities of PLLs were assessed by measuring lactate dehydrogenase (LDH) release from HTSE cel1s during treatment. The results were as follows: PLLs significantly inhibited basic mucin release from cultured HTSE cells in a dose-dependent manner from the range of 46mer to 14mer; PLL 46mer significantly inhibited the stimulated mucin release by ATP from cultured HTSE cells; there was no significant release of LDH from cultured HTSE cells during treatment. We conclude that PLLs inhibit both physiological and stimulated mucin release from airway epithelial cells without significant cytotoxicity and PLL lost its activity under the range of 14mer. This finding suggests that polymer of basic amino acid like PLL might function as a regulator for hypersecretion of mucus manifested in various respiratory diseases.

• 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.27 no.1
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• pp.126-137
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• 2006

In the present study, the author intended to investigate whether three oriental medical prescriptions named Jeongcheonhwadam-tang(JHT), Haengso-tang(HST), Socheungryong-tang(SCRT) significantly affect mucin release from cultured hamster tracheal surface epithelial (HTSE) cells. The results were as follows: (1) JHT significantly inhibited mucin release from cultured HTSE cells, without significant cytotoxicity : (2) HST significantly inhibited mucin release from cultured HTSE cells, without significant cytotoxicity : (3) SCRT significantly inhibited mucin release from cultured HTSE cells, without significant cytotoxicity : (4) JHT, HST chiefly inhibited the 'mucin' release and did not significantly affect the release of the other releasable glycoproteins with less molecular weight than mucin. These results suggest that the three herbal prescriptions specifically inhibit the release of mucin: (5) JHT significantly inhibited the expression levels of MUC SAC mRNA. This result suggests that JHT affects the synthesis of mucin at gene level in cultured HTSE cells. All agents showed no significant cytotoxicity. In view of these results, further investigation of the effects of JHT and HST are likely to be instrumental in yielding novel agents from oriental medical prescriptions which have inhibitory effects or expectorative effects on airway mucus secretion.

• Biomolecules & Therapeutics
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• v.12 no.1
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• pp.1-8
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• 2004

In chronic airway inflammatory diseases such as asthma and chronic bronchitis, it has been suggested that matrix metalloproteinases secreted from infiltrating neutrophil contribute the pathogenesis of the disease and have been a focus of intense investigation. We report here that hamster tracheal surface epithelial goblet cells (HTSE cells) produce matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2). Matrix metalloproteinase activities were investigated using [$^3H$]collagen-digestion assay and gelatin zymography. The subtype of matrix metalloproteinases expressed from HTSE cells was MMP-2 (gelatinase A), which was determined by Western blot with various subtype selective anti-matrix metalloproteinase antibodies. The MMP-2 and TIMP-2 cDNAs from HTSE cells were partially cloned by RT-PCR and they reveal more than 90% of sequence homology with those from human, rat and mouse. The collagenolytic activity was increased with the secretory differentiation of the HTSE cell and it was found that zymogen activation was responsible for the increased MMP-2 activity in HTSE cells. The results from the present study suggest that the metaplastic secretory differentiation of airway goblet cells may affect chronic airway inflammatory process by augmenting the zymogen activation of MMP-2.

• The Journal of Internal Korean Medicine
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• v.27 no.1
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• pp.92-101
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• 2006

Objectives : This study was done with intend to investigate whether two oriental medical prescriptions, daecheongryong-tang (DCRT) and prescription A (P-A) significantly affect mucin 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 DCRT or P-A 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 DCRT and P-A on contractility of isolated tracheal smooth muscle were investigated. Results were as follows : 1. DCRT significantly inhibited mucin release from cultured HTSE cells, with significant cytotoxicity. 2. P-A significantly increased mucin release from cultured HTSE cells, with significant cytotoxicity. 3. DCRT inhibited Ach-induced contraction of isolated tracheal smooth muscle. 4. P-A also inhibited Ach-induced contraction of isolated tracheal smooth muscle. Conclusion: Results suggest that DCRT and P-A have regulating effects on mucin secretion from goblet cells. Further investigation is needed, because of the value in finding novel agents to this purpose, and these oriental medical prescriptions have potential for this role.

• 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.

• The Journal of Pediatrics of Korean Medicine
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• v.19 no.1
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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.

• The Journal of Internal Korean Medicine
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• v.27 no.4
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• pp.895-904
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• 2006

Objectives : In the present study, the author intended to investigate whether two oriental medical prescriptions named Socheongryongtang-ga-seoggo (SCTS) and Prescription D (P-D) significantly affect mucin release from cultured hamster tracheal surface epithelial (HTSE) cells. Materials and Methods : Confluent HTSE cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of SCTS or P-D 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 SCTS and P-D on contractility of isolated tracheal smooth muscle were investigated. Results : SCTS did not affect mucin release from cultured HTSE cells, without cytotoxicity. However, P-D significantly increased mucin release from cultured HTSE cells. with significant cytotoxicity. SCTS inhibited Ach-induced contraction of isolated tracheal smooth muscle. P-D also inhibited Ach-induced contraction of isolated tracheal smooth muscle. Conclusions : The author suggests that the effects of SCTS and P-D with their components should be further investigated and it is valuable to find, from oriental medical prescriptions, novel agents which might regulate mucin secretion from airway goblet cells.

• The Journal of Pediatrics of Korean Medicine
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• v.21 no.3
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• pp.33-55
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• 2007

Objectives In the present study, the author intended to investigate whether bojung-ikgitang-gamibang(BJGB) and seonbang-paedoktang(SBPT) significantly affect in vivo and in vitro mucin secretion from airway epithelial cells. Methods In vivo experiment, mice's mucin which is on a hypersecretion of airway mucin, mice's tracheal goblet cells in hyperplasia and mice's intraepithelial mucosubstances were exposed with SO2for3weeks. Effects of orally-administered BJGB and SBPT during 1 week on vivo mucin secretion and hyperplasia of tracheal goblet cells were assessed by using both enzyme-linked immunosorbent assay(ELISA) and staining goblet cells with alcian blue. In vitro experiment, confluent hamster tracheal surface epithelial(HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24hrs and chased for 30 min in the presence of each agent to figure out the effectiveness of 3H-mucin secretion. Total elution profiles of control spent media and treatment sample through Sepharose CL-4B column were analyzed. The effects of each agent on contractility of isolated tracheal smooth muscle and effects of each agent on MUC5AC gene expression in cultured HTSE cells were investigated. Also, possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase(LDH) release. Additionally, effects of BJGB and SBPT on both MUC5AC gene expression in cultured HTSE cells and TNF- or EGF-induced MUC5AC gene expression in human airway epithelial cells (NCI-H292) were investigated. Results (1) BJGB and SBPT inhibited hypersecretion of in vivo mucin. SBPT also inhibited the increase the number of goblet cells. However, BJGB did not affect the increase of number of goblet cells; (2) BJGB significantly increased mucin secretion from cultured HTSE cells, without significant cytotoxicity, and chiefly affected the 'mucin' secretion; (3) SBPT did not affect mucin secretion from cultured HTSE cells without significant cytotoxicity, and also did not affect the secretion of the other releseable glycoproteins; (4) BJGB and SBPT did not affect Ach-induced contraction of isolated tracheal smooth muscle; (5) SBPT significantly inhibit the expression levels of MUC5AC gene and BJGB significantly increased the expression levels of MUC5AC gene in both HTSE cells and NCI-H292 cells. Conclusions BJGB and SBPT can not only affect the secretion of mucin but also affect the expression of mucin gene. The author suggests that the effects BJGB and SBPT with their components should be further investigated and it is highly desirable to find from oriental medical prescriptions, novel agents which might regulate hypersecretion of mucin from airway epithelial cells.