• Biomolecules & Therapeutics
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• v.13 no.4
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• pp.251-255
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• 2005

In the present study, we tried to investigate whether protein kinase C (PKC) activator, phorbol 12-Myristate 13-Acetate (PMA), and PKC inhibitors, $G\"{o}-6976$, Ro-31-8220 and rottlerin significantly affect basal mucin relesed 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 in the presence of each agent to assess the effects on $^3H$-mucin release. The results were as follows: (1) PMA increased mucin release from cultured HTSE cells, during 30 min of treatment period; (2) However, $G\"{o}-6976$, Ro-31-8220 and rottlerin did not significantly affect mucin release, during 30 min of treatment period. This finding suggests, at least in part, that PKC might playa minor role in the signaling pathways involved in basal - physiological or constitutive - mucin release from airway goblet cells, although further studies are needed.

• 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.

• Biomolecules & Therapeutics
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• v.9 no.3
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• pp.218-223
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• 2001

In the present study, we intended to investigate whether polycationic peptides including poly-L-lysine (PLL) and poly-L-arginine (PLA) specifically inhibit the mucin release and do not affect significantly the release of the other releasable glycoproteins with less molecular weight than mucin's 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 in the presence of varying concentrations of either poly-L-arginine (PLA) or poly-L-lysine (PLL) to assess the effects on 3H-mucin release and on the total elution profile of the treated culture medium. The results were as follows : (1) PLL 78,000, PLL 9,600 and PLA 8,900 inhibited mucin release in a dose-dependent manner; (2) These polycationic peptides did not inhibit the release of the other releasable glycoproteins with less molecular weights than mucin's. We conclude that these polycationic peptides 'specifically'inhibit mucin release from airway goblet cells. This finding suggests that these polycationic peptides might be used as a specific airway mucin-regulating agent.

• Journal of dental hygiene science
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• v.2 no.1
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• pp.25-29
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• 2002

In the present study, we tried to investigate whether poly-L-lysine(PLL)(MW 78,000 and 9,600) significantly affect mucin release from cultured hamster 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 in the presence of varying concentrations of PLL to assess the effects on $^3H$-mucin release. Possible cytotoxicities of PLL were assessed by measuring Lactate Dehydrogenase(LDH) release during treatment. The results were as follows : (1) PLL significantly inhibited mucin release from cultured HTSE cells in a dose-dependent manner; (2) there was no significant release of LDH by treatment of PLL 9,600; (3) however, in the case of treatment of PLL 78,000, there was significant release of LDH during treatment. We conclude that PLL which has molecular weight under 10,000 might inhibit mucin release from airway goblet cells without significant cytotoxicity. This finding suggests that PLL might be used as a tool of research for the hypersecretion of airway mucus.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1995.04a
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• pp.89-89
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• 1995

Surface epithelial cells isolated from hamster tracheas and grown on a thick collagen gel become a highly enriched population of mucus-secreting cells. Epithelial cells from tracheas of hamsters were collected using enzymatic procedures and cultured under various conditions. The medium used consisted of a 1:1 mixture of medium 199 and Dulbecco's modified Eagle's (DME) medium which was conditioned before use. Insulin, transferrin, hydrocortisone, epidermal growth factor, and extract from bovine hypothalamus were used as supplement. Due to relatively low basal rates of min secretion from in vitro cultures, cultures are generally radiolabeled using $^3$H-glucosamine as a metabolic precursor. The radiolabeled mucinsreleased are quantitated by precipitation with TCA/PTA. Using this cell culture system, we investigated mucin release of goblet cells by altering the media bathing the apical surface of hamster tracheal surface epithelial(HTSE) cells. Acidic media added sulfuric acid caused sigcificant increases in mucin relesse (155${\pm}$20% at pH 4 and 146${\pm}$16% at, pH 5). Ammonium hydroxide also increased mucin release at pH 9.0(156${\pm}$17%) and pH 10(295${\pm}$9%) respectively. This additional mucin release seems to be associated with cell membrane damage as indicated by release of cellular LDH. SP stimulates secretion of mucin in cultured HTSE cells(154${\pm}$16% at 1${\times}$10$\^$-6/M and 165${\pm}$25% at 1${\times}$10$\^$-5/M. PAF at 5${\times}$10$\^$-6/M and 5${\times}$10$\^$-5/M enhanced by HTSE cells in vitro 168${\pm}$34% and 259${\pm}$30% of mucin secretion, respectively. The increase in mucin release by PAF and SP was not secondary to cell damage or necrosis. SP and PAF may be in mediating mucous secretion induced by inflammation irritantion and infection.

• The Journal of Pediatrics of Korean Medicine
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• v.21 no.1
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• pp.117-137
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• 2007

Objectives : In the present study, the author intended to investigate Gagam-jeonggitang(GJT), Gami-hwajeongjeon(GHJ) and Gami-tonggyutang(GTT) significantly affect in vivo and in vitro mucin secretion from airway epithelial cells. Methods : In vivo experiment, the author induced hypersecretion of airway mucin, hyperplasia of tracheal goblet cells and the increase in intraepithelial mucosubstances by exposing rats to SO2 during 3 weeks. Effects of orally-administered GJT, GHJ and GTT during 1 week on in vivo mucin secretion and hyperplasia of tracheal goblet cells were assesed using ELISA and staining goblet cells with alcian blue. For in vitro experiment, confluent HTSE cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of each agent to assess the effects of each agent on 3H-mucin secretion. Possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase release. Also, 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. Results : GJT, GHJ and GTI inhibited hypersecretion of in vivo mucin: GJT and GHJ inhibited the increase of number of goblet cells. However, GTT did not affect the increase of number of goblet cells; GJT and GTT significantly increased mucin secretion from cultured HTSE cells, without significant cytotoxicity. GHJ increased mucin secretion and showed mild cytotoxicity at the highest concentration: GJT, GHJ and GTT chiefly affected the 'mucin' secretion; GJT, GHJ and GTT did not affect Ach-induced contraction of isolated tracheal smooth muscle; GTT did not significantly affect the expression levels of MUC5AC gene. However, GJT significantly. inhibit the expression levels of MUC5AC gene and GHJ significantly increased the expression levels of MUC5AC gene. These results suggest that GJT, GHJ and GTI can increase mucin secretion during short-term treatment(in vitro), whereas it can inihibit hypersecretion of mucin during long-term treatment(in vivo) and GJT and GHJ can not only affect the secretion of mucin but also affect the expression of mucin gene. Conclusions : The author suggests that the effects GJT, GHJ and GTT with their components should be further investigated and it is valuable to find, from oriental medical prescriptions, novel agents which might regulate hypersecretion of mucin from airway epithelial cells.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.04a
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• pp.219-219
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• 1996

Mucin release from hamster tracheal surface epithelial(HTSE) cells can be stimulated by extracellular ATP via activation of P$_2$ purinoceptors located on the cell surface which appears to be coupled to phospholipase C via G proteins. However, our preliminary data indicate that the ATP-induced mucin release involves, in part, activation of PKC, but not an increase in the intracellular Ca++ level, suggesting the presence of another pathway which is separate from the PLC-PKC pathway, In this study, we intended to confirm the previous observation and subsequently identify an additional mechanism. Confluent HTSE cells were metabolically labeled with either $^3$H-glucosamine or $^3$H-arachidonic acid(AA), and release of either $^3$H-mucin or $^3$H-AA was quantified following various treatments. $^3$H-mucin was assayed using the sepharose CL-4B gel-filtration method, whereas $^3$H-AA liberation was measured by counting $^3$H-radioactivity in the chase medium. We found that: (1)Desensitization of PKC by pretreatment with PMA completely abolished the mucin releasing effect of PMA but partially inhibited the ATP-induced mucin release; (2) ATP increases release of $^3$H-AA in a dose-dependent fashion; (3) mepacrine, an inhibitor of PLA$_2$, attenuates ATP-induced mucin release in a dose-dependent fashion. These results confirm our previous notion that the PLC-PKC pathway is responsible, in part, for ATP-induced mucin release. Furthermore, activation of PLA$_2$ appears to be an additional pathway which is involved in ATP-induced mucin release.

• Transactions of the Korean hydrogen and new energy society
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• v.20 no.5
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• pp.432-438
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• 2009

In this paper, we synthesized LSG powder by Modified-GNP method. Lanthanum, strontium and gallium (LSG) were selected in the preparation of an oxygen-electrode (anode) for High Temperature Steam Electrolysis system (HTSE). The used amount and concentration of nitric acid were varied to find out an appropriate composition for oxygen-electrode (anode). In order to optimize the molar ratio of La and Sr, ratio of La to Sr was varied that 2:8, 5:5 and 8:2. The combined LSGs were calcined for 2 hours at $700^{\circ}C$ and were sintered in a furnace for 4 hours at $1200^{\circ}C$. The phase and crystallinity of LSG powder were determined by XRD. The surface morphology was observed through SEM photograph, and the specific surface area was investigated with BET. The thermochemical property was determined by TG/DTA. The synthesized preparation was obtained of $La_{0.8}Sr_{0.2}GaO_{3}$ formula for 3M nitric acid, which was the best perovskite phase.

• The Korean Journal of Physiology and Pharmacology
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• v.7 no.3
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• pp.143-147
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• 2003

Poly-L-lysine (PLL) was reported to suppress mucin release from airway goblet cells during 30 min treatment period. In this study, we investigated whether PLL consistently suppresses mucin release from cultured airway goblet cells during 24 h after 30 min treatment and also specifically suppresses the release of mucin without any effects on the other releasable glycoproteins. Confluent primary hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with $^3H$-glucosamine for 24 h and chased for 30 min in the presence of varying concentrations of PLL to assess the effects on $^3H$-mucin release and on the total elution profile of the treated culture medium. The total mucin content during 24 h after 30 min treatment of PLL was assesed to investigate the consistency of effects. PLL did not affect the release of the other releasable glycoproteins whose molecular weights were less than mucin, and decreased the total mucin content during 24 h after 30 min treatment. We conclude that PLL can specifically suppress mucin release from cultured airway goblet cells and the suppression on mucin release is consistent. This finding suggests that PLL might be used as a specific airway mucin-regulating agent by directly acting on airway mucin-secreting cells.

• The Journal of Pediatrics of Korean Medicine
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• v.21 no.1
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• pp.139-154
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• 2007

Objectives : The author intended to investigate Seonbangpaedoktang (SBPT) significantly affect in vivo and in vitro mucin secretion from airway epithelial cells. Methods : In vivo experiment, the author induced hypersecretion of airway mucin, hyperplasia of tracheal goblet cells and the increase in intraepithelial mucosubstances. Effects of orally-administered SBPT during 1 week on in vivo mucin secretion and hyperplasia of tracheal goblet cells were assessed. For in vitro experiment, confluent hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled and chased in the presence of SBPT 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 analysed. Possible cytotoxicity of the agent was assessed by measuring LDH release. Also, the effect of SBPT on contractility of isolated tracheal smooth muscle was investigated. Results : SBPT inhibited hypersecretion of in vivo mucin and inhibited the increase of number of goblet cells ; SBPT did not affect in vitro mucin secretion and the secretion of the other releasable glycoproteins with less molecular weight than mucin from cultured HTSE cells, without significant effect on LDH release; SBPT did not affect Ach-induced contraction of isolated tracheal smooth muscle. Conclusions : SBPT can inihibit hypersecretion of in vivo mucin and the author suggest that the effect SBPT with their components should investigate further.