• Asian-Australasian Journal of Animal Sciences
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• 제18권5호
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• pp.716-722
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• 2005

Carcass weight and backfat thickness are primary yield grading factors. Insulin-like growth factor (IGF)-I/-II, transforming growth factor $\beta$1 (TGF-$\beta$1), and epidermal growth factor (EGF) regulate the proliferation and differentiation of cells including adipocytes. Also, interleukin (IL)-2/-6, cortisol, and dehydroepiandrosterone-sulfate (DHEA-S) are known to be related to muscle growth and fat depth. However, the relationships between endocrine factors and carcass grade have not been studied. Therefore, this study aimed to measure the concentrations of endocrine factors in serum and muscle, and to investigate the relationship of endocrine factors with carcass grade. A total of 60 crossbred gilts (Duroc${\times}$Yorkshire${\times}$Landrace) were used. Blood from the jugular vein was collected at antemortem (7 days before slaughter) and postmortem periods, and M. Longissimus was collected at 45 min and 24 h after slaughter. The concentrations of IGF-I/-II, EGF, TGF-$\beta$1, IL-2/-6, cortisol and DHEA-S were analyzed by radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA). In general, IGF and EGF concentrations in serum and muscle of grade A carcasses were found to be higher than those of grade C carcasses at antemortem and postmortem periods, whereas the pattern of TGF-$\beta$1 concentration was reversed. In particular, the concentrations of muscle IGF-I (24 h postmortem) and serum TGF-$\beta$1 (antemortem) were significantly different between grades A and C (p<0.05). The present results indicate that serum and muscle growth factors affect carcass weight and backfat thickness, and indirectly suggest the possibility that carcass grade could be predicted by expression of serum and/or muscle growth factors.

• Natural Product Sciences
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• 제22권4호
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• pp.275-281
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• 2016

Perilla frutescens was empirically used for controlling airway inflammatory diseases in folk medicine. We investigated whether caffeic acid, myristicin and rosemarinic acid derived from Perilla frutescens significantly affect the gene expression and production of mucin from airway epithelial cells. Confluent NCI-H292 cells were pretreated with caffeic acid, myristicin or rosemarinic acid for 30 min and then stimulated with phorbol 12-myristate 13-acetate (PMA) for 24 h. The MUC5AC mucin gene expression and production were measured by reverse transcription - polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. Additionally, we examined whether caffeic acid, myristicin or rosemarinic acid affects MUC5AC mucin production indued by epidermal growth factor (EGF) and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), the other two stimulators of production of airway mucin. The results were as follows: (1) Caffeic acid, myristicin and rosemarinic acid inhibited the gene expression and production of MUC5AC mucin induced by PMA from NCI-H292 cells, respectively; (2) Among the three compounds derived from Perilla frutescens, only rosemarinic acid inhibited the production of MUC5AC mucin induced by EGF or $TNF-{\alpha}$, the other two stimulators of production of airway mucin. These results suggest that rosemarinic acid derived from Perilla frutescens can regulate the production and gene expression of mucin, by directly acting on airway epithelial cells and, at least in part, explains the traditional use of Perilla frutescens as remedies for diverse inflammatory pulmonary diseases.

• The Korean Journal of Physiology and Pharmacology
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• 제22권6호
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• pp.671-677
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• 2018

In the present study, we investigated whether tussilagone, a natural product derived from Tussilago farfara, significantly affects the production and gene expression of airway MUC5AC mucin. Confluent NCI-H292 cells were pretreated with tussilagone for 30 min and then stimulated with EGF (epidermal growth factor) or PMA (phorbol 12-myristate 13-acetate) for 24 h or the indicated periods. The MUC5AC mucin gene expression was measured by RT-PCR. Production of MUC5AC mucin protein was measured by ELISA. To elucidate the action mechanism of tussilagone, effect of tussilagone on PMA-induced $NF-{\kappa}B$ signaling pathway was investigated by western blot analysis. Tussilagone significantly inhibited the production of MUC5AC mucin protein and down-regulated the expression of MUC5AC mucin gene, induced by EGF or PMA. Tussilagone inhibited PMA-induced activation (phosphorylation) of inhibitory kappa B kinase (IKK), and thus phosphorylation and degradation of inhibitory kappa Ba ($I{\kappa}B{\alpha}$). Tussilagone inhibited PMA-induced phosphorylation and nuclear translocation of nuclear factor kappa B ($NF-{\kappa}B$) p65. This, in turn, led to the down-regulation of MUC5AC protein production in NCI-H292 cells. These results suggest that tussilagone can regulate the production and gene expression of mucin by acting on airway epithelial cells through regulation of $NF-{\kappa}B$ signaling pathway.

• Tuberculosis and Respiratory Diseases
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• 제78권3호
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• pp.210-217
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• 2015

Background: Adenophora triphylla var. japonica is empirically used for controlling airway inflammatory diseases in folk medicine. We evaluated the gene expression and production of mucin from airway epithelial cells in response to lupenone, lupeol and taraxerol derived from Adenophora triphylla var. japonica. Methods: Confluent NCI-H292 cells were pretreated with lupenone, lupeol or taraxerol for 30 minutes and then stimulated with tumor necrosis factor ${\alpha}$ (TNF-${\alpha}$) for 24 hours. The MUC5AC mucin gene expression and production were measured by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Additionally, we examined whether lupenone, lupeol or taraxerol affects MUC5AC mucin production induced by epidermal growth factor (EGF) and phorbol 12-myristate 13-acetate (PMA), the other 2 stimulators of airway mucin production. Results: Lupenone, lupeol, and taraxerol inhibited the gene expression and production of MUC5AC mucin induced by TNF-${\alpha}$ from NCI-H292 cells, respectively. The 3 compounds inhibited the EGF or PMA-induced production of MUC5AC mucin in NCI-H292 cells. Conclusion: These results indicated that lupenone, lupeol and taraxerol derived from Adenophora triphylla var. japonica regulates the production and gene expression of mucin, by directly acting on airway epithelial cells. In addition, the results partly explain the mechanism of of Adenophora triphylla var. japonica as a traditional remedy for diverse inflammatory pulmonary diseases.

• IMMUNE NETWORK
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• 제12권4호
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• pp.155-164
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• 2012

It is well established that blocking the interaction of EGFR with growth factors leads to the arrest of tumor growth, resulting in tumor cell death. ER414 is a human monoclonal antibody (mAb) derived by guided selection of the mouse mAb A13. The ER414 exhibited a ~17-fold lower affinity and, as a result, lower efficacy of inhibition of the EGF-mediated tyrosine phosphorylation of EGFR when compared with mAb A13 and cetuximab. We performed a stepwise in vitro affinity maturation to improve the affinity of ER414. We obtained a 3D model of ER414 to identify the amino acids in the CDRs that needed to be mutated. Clones were selected from the phage library with randomized amino acids in the CDRs and substitution of amino acids in the HCDR3 and LCDR1 of ER414 led to improved affinity. A clone, H3-14, with a ~20-fold increased affinity, was selected from the HCDR3 randomized library. Then three clones, ER2, ER78 and ER79, were selected from the LCDR1 randomized library based on the H3-14 but did not show further increased affinities compared to that of H3-14. Of the three, ER2 was chosen for further characterization due to its better expression than others. We successfully performed affinity maturation of ER414 and obtained antibodies with a similar affinity as cetuximab. And antibody from an affinity maturation inhibits the EGF-mediated tyrosine phosphorylation of EGFR in a manner similar to cetuximab.

• 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.

• 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.

• 대한약리학회지
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• 제29권1호
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• pp.139-148
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• 1993

SCC 12와 SCC 13 세포주는 피부의 상피암 조직에서 유래되었다. 본 연구에서는, 세포배양으로 in vivo 피부의 거의 모든 특징들을 재현할 수 있는 raft 배양법을 이용하여, 두 종류의 상피 암세포주의 진피 침투성 정도를 비교해 보았다. SCC 12 세포주의 raft 배양에서는, collagen matrix로 침투하는 많은 세포군들이 관찰되었고, 또한 기저세포와 유사한 세포들이 분화층 중간에 관찰되었다. 그러나 SCC 13 세포주의 raft 배양에서는 침투성 세포들이 전혀 관찰되지 않았다. 어떤 인자가 SCC 12 세포주의 침투성에 관여하였는지를 조사하기 위하여, 세포이동 또는 matrix 파괴에 관여하는 각종 단백질 즉, type 1 collagenase, Fn, Fn 수용체와 EGF 수용체 등의 기본 생합성속도를 침투성과 비침투성 세포주에서 측정하였다. 그 중 type 1 collagenase의 생합성만이 침투성 세포주에서 현저히 높게 나타났다. 침투성 SCC 12 세포주에서 관찰된 높은 type 1 collagenase 생합성과 이들 세포주의 침투성과의 연관관계 및 그 기작을 조사하기 위하여 두 세포주에서 TGF alpha에 대한 반응의 차이를 조사하였다. 비침투성 세포주에서는, TGF alpha에 의해 type 1 collagenase의 생합성이 처음 12시간 동안에 현저한 증가를 보였으나 24시간부터 그 유도가 down-regulation되는 현상을 보였다. 그러나 침투성 세포는 TGF alpha에 의해 계속적인 type 1 collagenase의 유도를 보였다. 이에 저자들은 EGF 수용체의 down-regulation으로 인해 type 1 collagenase의 down-regulation이 유도되고, 이와 같은 regulation의 결함으로 인해 SCC 12 세포주가 collagen matrix로 침투하는 현상을 나타낼 것으로 제시하였다.

• 대한한방내과학회지
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• 제29권2호
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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.

• 대한한의학회지
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• 제31권1호
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• pp.138-152
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• 2010

Objective: Yikgeebohyul-tang (Yiqib$\check{u}$xu$\grave{e}$-t$\bar{a}$ng) is one of the representative prescriptions for invigoration of vitality and nourishing of the blood. This study was carried out to investigate the effects of Yikgeebohyul-tang (Yiqib$\check{u}$xu$\grave{e}$-t$\bar{a}$ng) on hair regrowth and cytokine changes in a shaving model of C57BL6 mice. Method: Five-week-old mice were acclimated for 1 week at a temperature between $21-23^{\circ}C{\acute{E}}$, 40-60% relative humidity, and 12h of a light/dark cycle before beginning of the experiment. There were two groups including normal saline (control) and a positive control of oral intake of Yikgeebohyul-tang (Yiqib$\check{u}$xu$\grave{e}$-t$\bar{a}$ng) extract (sample) in 18 female mice. The test compounds were topically treated once a day over 14 days. The hair regrowth was photographically and histologically determined during the experimental period of 14 days. Revelation of TGF-${\beta}1$ and EGF were also determined using immunohistochemistry. In addition to that, IFN-$\gamma$, IL-4 and IL-10 were determined in serum. Results: Hair regrowth in the sample group was promoted earlier and faster than the control group, as shown by concentrations of hairs and thick-hair ratio in the sample group. TGF-${\beta}1$ was not revealed in either control or sample group. EGF was strongly positive in out root sheath of some thick hair of the sample group. Serum IFN-$\gamma$ was significantly decreased in the sample group compared with the control group at 7 experimental days. However, it was not significantly decreased at 14 experimental days. Serum IL-4 was significantly increased in the sample group compared with the control group at 7 experimental days. However, it was not significantly decreased at 14 experimental days. Serum IL-10 was decreased in the sample group compared with the control group, but with no real statistical significance. Conclusion: These results suggest that Yikgeebohyul-tang (Yiqib$\check{u}$xu$\grave{e}$-t$\bar{a}$ng) has hair growth promoting activity and it can be used for treatment of alopecia. Also, these effects relate to EGF revelation of hair roots, a decrease in serum IFN-$\gamma$, and an increase of serum IL-4.