• The Korean Journal of Physiology and Pharmacology
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• v.2 no.6
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• pp.763-770
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• 1998

A number of substances involved in the proliferation and differentiation of the tracheobronchial epithelium have been identified. The defects in the control of the proliferation and differentiation of tracheobronchial epithelial cells appear to constitute crucial steps in the transition of normal cells to neoplastic ones. Endothelin-1 is produced by tracheal epithelial cells, and its receptors are present in tracheal epithelial cells. However, the effect of endothelin-1 on the proliferation and differentiation of tracheal epithelial cells has not been clearly elucidated. This study was undertaken to investigate these actions of endothelin-1 in primary cultured cells of rat tracheal epithelia. Endothelin-1 stimulated proliferation of tracheal epithelial cells 1.5-fold when compared with that of control cells. Endothelin-1 increased mitogen-activated protein kinase (MAPK) activity. Herbimycin A, a tyrosine kinase inhibitor, inhibited endothelin-1-induced proliferation of epithelial cells. The treatment of endothelin-1 during the primary culture of tracheal epithelial cells increased AB-PAS-stained cell population and ciliated cell population 6.5 fold and 1.5 fold, respectively, when compared with those in control cells. The responsiveness to carbachol and forskolin in the $Cl^-$ secretion was increased 1.7 and 1.9 fold, respectively, in the endothelin-treated epithelial cells. These results indicated that endothelin-1 increases proliferation via MAPK pathway and stimulates differentiation to secretory and ciliated cells in rat tracheal epithelial cells.

• Biomolecules & Therapeutics
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• v.10 no.4
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• pp.281-286
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• 2002

In this study, we intended to get a preliminary data for establishing rat tracheal surface epithelial(RTSE) cell culture system as an experimental model for physiology and pharmacology of tracheal epithelial cells. Primary culture on the membrane support and application of the air-liquid interface system at the level of cell layer were performed. The cell growth rate and mucin production rate were measured according to the days in culture. The results were as follows: this culture system was found to manifest mucocilliary differentiation of rat tracheal epithelial cells, the cells were confluent and the quantity of produced and released mucin was highest on culture day 9, the mucin was mainly released to the apical side and tbe free $^3{H}$-glucosamine which was not incorporated to process of synthesis of mucin was left on the basolateral side. Taken together, we suggest that air-liquid interface culture system can be used as a substitute for immersion culture system and as an experimental model for in vivo mucus-hypersecretory diseases.

• Proceedings of the Korean Biophysical Society Conference
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• 1997.07a
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• pp.22-22
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• 1997

The physiological activity of tracheal epithelial cells is closely related with the ionic conditions of cytosol, specially the concentration of cytosolic Ca$\^$2＋/. We have prepared microsomes in these cells and the molecular mechanisms of ionic regulations were investigated.(omitted)

• 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.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.

• The Journal of Korean Medicine
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• v.28 no.2 s.70
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• pp.54-65
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• 2007

Objectives : In the present study, the author investigated whether Naenghyo-hwan(NHH) significantly affect 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 NHH to assess the effect of the agent on 3H-mucin secretion. Total elutionprofiles of control spent media and treatment sample through Sepharose CL-4B column were analysed. Effect of NHH on contractility of isolated tracheal smooth muscle was investigated. Also, effect of the agent on MUC5AC gene expression in cultured NCI-H292cells was investigated. Possible cytotoxicities of the agent were assessed by measuring both lactate dehydrogenase (LDH) release from HTSE cells and examining the rate of survival and proliferation of NCI-H292 cells. Results : NHH significantly increased mucin secretion from cultured HTSE cells, with significant cytotoxicity. NHH chiefly affected the 'mucin' secretion. NHH inhibited ACh-induced contraction of isolated tracheal smooth muscle. NHH disturbed both the extraction of total RNA from NCI-H292 cells and polymerase chain reaction, nonspecifically. Therefore, in this experiment, theeffect of NHH on the expression levels of MUC 5AC gene in cultured NCI-H292 cells could not be elucidated. Conclusions : The author suggests that the effect of NHH 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 epithelial cells.

• Korean Journal of Bronchoesophagology
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• v.8 no.1
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• pp.29-34
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• 2002

Background and Objectives : Sulfur dioxide gas is one of the major airborne Pollutants noxious to human in industrialized countries. The most vulnerable areas in the human respiratory system were the trachea and main bronchi and a gradient of decreasing damage was observed in the peripheral tracheobronchial tree. Induced functional alteration was increased mucosal permeability, and morphological changes were epithelial sloughing, intracellular edema, mitochondrial swelling, widened intercellular spaces, and ciliary cytoplamic extrusions. The laminins are a family of extracellular matrix glycoproteins localized in the basement membrane. Their primary role is cell-matrix attachment, but many additional biologic activities, including Promoting cell growth and migration, tumor growth and metastasis, wound repair, and graft survival, have been demonstrated. Materials and Methods : Histologic changes and expression of laminin in tracheal mucosa sacrificed at 1 day, 2 day, 3 day, 1 week, 2 weeks, and 3 weeks after continued SO2 exposure of 250 ppm for 30 minutes a day(to 7week) were studied in rats. In this study, mild immune reaction for laminin was noted at the apical cytoplasm of epithelial cells and basement membrane one day after a 7 week $SO_2$ exposure. The cilia and nucleoi of epithelial cells were normal and no immune reaction was noted in Goblet cells. The lamina propria of the tracheal tissue was infiltrated by monocytes and lymphocytes. Results : At 24 hours after exposure, all tracheal cells except Goblet cells revealed a mild immune reaction for laminin. No immune reactions were noted in the basement membrane. At 72 hours after exposure, mild or moderate immune reactions for laminin was seen in the tracheal cell cytoplasm. Irregular faint immune reaction for laminin was noted in the basement membrane. At 1 week after exposure, strong immune reaction for laminin was detected over all tracheal cells, and the basement membrane was seen clearly. At 2~3 weeks after exposure, strong immune reaction for laminin was seen in all tracheal epithelial cells except Goblet cells and a mild immune reaction was partly revealed in the basement membrane. Conclusion : Our study suggests that 502 produces histologic damage on the tracheal mucosa. Longer duration after exposure of $SO_2$ makes more progressive healing on the tracheal mucosa and increased immunoreactivity for laminin.

• Korean Journal of Veterinary Research
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• v.37 no.1
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• pp.87-101
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• 1997

The development of trachea in fetuses on 60, 90 and 120 days of gestation and neonates of Korean native goats was investigated by microscopy and scanning and transmission electron microscopy. The results were summarized as follows; Light microscopic studies: 1. In the 60-day-old fetuses, the tracheal walls were differentiated and divided into four layers of the mucosa, submucosa, muscle and cartilage, and adventitia. The tracheal epithelium is composed of stratified ciliated columnar epithelium at 60- and 90-day-old fetuses while the epithelium observed at 120-day-old fetuses was pseudostraified ciliated colummar epithelium. 2. In the 90-day-old fetuses, tracheal glands extended into the submucosa and peripheral area of the tracheal cartilage. The blood vessels were observed in the submucosa and adventitia. The elastic and collagenous fibers were observed in the tracheal walls. 3. In the neonates, the tracheal walls consisted of mucosa with well-developed folds, submucosa, tracheal glands, muscle and cartilage, collagenous and elastic fibers, and adventitia, which were more developed than those of 120-day-old fetuses. The tracheal epithelium was developed as that in adults. Scanning electron microscopic studies: 4. In the 60-day-old fetuses, most of tracheal epithelial cells were nonciliated but short microvilli were sporadically observed on the luminal surface. On rare occasions, a few cells have solitary cilium. 5. In the 90-day-old fetuses, the ciliated cells appeared increasingly and cilia elongated longer than those of 60-day-old fetuses. 6. In the 120-day-old fetuses, the nonciliated cells covered with microvilli in dome-shape were barriered by thick carpet of cilia. The nonciliated cells also have many papillary projectons on the apical surface. 7. In the neonates, the nonciliated cells in tracheal epithelium were covered compactly with numerous cilia, and many secretory droplets were found on the cilia. Transmission electron microscopic studies: 8. In the 60-day-old fetuses, nonciliated cells of the tracheal epithelium contain large amounts of glycogen granules in the supernuclear and subnuclear areas meanwhile a few cell organelles were formed. Cilia were well formed along the apical cell membranes of the ciliated cells. Also found in the ciliated cells were basal corpuscles, mitochondria and short chains in granular endoplasmic reticulum(GER). Between the epithelial cells presented were well-defined junctional complex with zonula occludens and desmosomes. The nuclei were variable in size and shape. The more developed nucleoli were observed conspicuosly. 9. In the 90-day-old fetuses, nonciliated cells contained large glycogen granules. Accumulated glycogen granules were observed in the subnuclear and supranuclear portion of the cytoplasm. A few short microvilli were covered with glycocalyx. Ciliated cells contained numerous mitochondria and short chains of GER. 10. In the 120-day-old fetuses, the ciliated cells contained numerous mitochondria, abundant short chains of GER and nucleoli. Nonciliated cells contained some Golgi complex and mitochondria. The cell borders were well-defined and distinct junctional complex with zonula occludens, desmosomes, and interdigitorum. 11. In the neonates, well-developed goblet cells were observed in the tracheal epithelium. Ultrastructures of ciliated and nonciliated cells on the tracheal epithelia were similar in pattern as those in adults.

• Journal of Chest Surgery
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• v.24 no.5
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• pp.438-444
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• 1991

This study was designed to evaluate the viability of canine trachea after cryopreservation for two months. Eight cervical tracheal rings were resected in three dogs and both ends were anastomosed. The resected tracheal segments were cryopreserved and stored in liquid nitrogen at -196oC for two months. Two months later, the cryopreserved segments were thawed. Half of each segment was implanted into the abdomen of its donor animal and the other half was cultured in tissue media. Two weeks later, the animal was sacrificed. The native cervical trachea was removed to serve as a control and the abdominally implanted trachea was removed for study. At that time, both specimens were also cultured in tissue media. Tracheal epithelial viability was assessed histologically by using an inverted microscope. The epithelial cells were confirmed immunohistochemically using monoclonal antibodies against cytokeratin and epithelial membrane antigen. Control and cryopreserved segments showed good, viable epithelial cells, but the implanted segments showed slightly depressed viability. We conclude that canine tracheal epithelium can survive after cryopreservation for two months, but the implanted trachea will be slightly damaged by ischemia before revascularization, even if omental wrapping is used.

• Proceedings of the Korean Biophysical Society Conference
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• 1998.06a
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• pp.29-29
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• 1998

Previously, we reported two types of vanadate-sensitive ATPases in the micro somes of tracheal epithelial cells, a high-affinity one and a low-affinity one. The low affinity vanadate-sensitive (LAVS) ATPase was sensitive to thapsigargin and cyclopiazonic acid, specific antagonists of ER-type Ca$\^$2＋/-ATPase, and mediated microsomal $\^$45/Ca$\^$2+/ uptake, implying that the LAVS-ATPase is an ER/SR-type Ca$\^$2+/-ATPase.(omitted)