• 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.

• 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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• 제21권1호
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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.

• 대한한방소아과학회지
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• 제21권3호
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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.

• Genes and Genomics
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• 제40권12호
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• pp.1383-1388
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• 2018

The development of therapeutic bacteriophages will provide several benefits based on an understanding the basic physiological dynamics of phage and bacteria interactions for therapeutic use in light of the results of antibiotic abuse. However, studies on bacteriophage therapeutics against microbes are very limited, because of lack of phage stability and an incomplete understanding of the physiological intracellular mechanisms of phage. The major objective of this investigation was to provide opportunity for development of a novel therapeutic treatment to control respiratory diseases in swine. The cytokine array system was used to identify the secreted cytokines/chemokines after Bordetella bronchiseptica infection into swine nasal turbinate cells (PT-K75). We also performed the real-time quantitative PCR method to investigate the gene expression regulated by B. bronchiseptica infection or bacteriophage treatment. We found that B. bronchiseptica infection of PT-K75 induces secretion of many cytokines/chemokines to regulate airway inflammation. Of them, secretion and expression of IL-$1{\beta}$ and IL-6 are increased in a dose-dependent manner. Interestingly, membrane-bound mucin production via expression of the Muc1 gene is increased in B. bronchiseptica-infected PT-K75 cells. However, cytokine production and Muc1 gene expression are dramatically inhibited by treatment with a specific B. bronchiseptica bacteriophage (Bor-BRP-1). The regulation of cytokine profiles in B. bronchiseptica-induced inflammation by B. bronchiseptica bacteriophage is essential for avoiding inappropriate inflammatory responses. The ability of bacteriophages to downregulate the immune response by inhibiting bacterial infection emphasizes the possibility of bacteriophage-based therapies as a novel anti-inflammatory therapeutic strategy in swine respiratory tracts.

• Tuberculosis and Respiratory Diseases
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• 제84권3호
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• pp.200-208
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• 2021

Background: Hypersensitivity pneumonitis (HP) is an increasingly recognized form of diffuse parenchymal lung disease. Krebs von den Lungen-6 (KL-6) is now classified as a human MUC1 mucin protein, and regenerating type II pneumocytes are the primary cellular source of KL-6/MUC1 in the affected lungs of patients with interstitial lung diseases (ILD). Serum KL-6/MUC1 levels have been demonstrated to be useful for the evaluation of various ILD. To determine the role of circulating KL-6 in evaluating the disease activity and management of HP. Methods: An observational cross-sectional study was conducted on 51 patients with HP and 20 healthy controls. Serum KL-6 levels were measured in both groups. Patients were further assessed based on chest high-resolution computed tomography (HRCT), pulmonary function test, 6-minute walk test, echocardiography, bronchioalveolar lavage, and/or transbronchial biopsy. Patients were divided into the fibrotic and non-fibrotic groups according to the HRCT findings. Results: The median serum KL-6 levels were significantly higher in HP patients as compared to the control group. The median serum KL-6 levels were found to be higher in the non-fibrotic HP group (1,900 IU/mL) as compared to the fibrotic group (1,200 IU/mL). There was a significant inverse correlation between serum KL-6 serum level and the dose of steroids as well as the duration of steroid therapy. Conclusion: The presence of higher KL-6 levels in the non-fibrotic HP group implies its enhanced production by regenerating pneumocytes in response to alveolar injury. The significant association between serum KL-6 levels and the dose and the duration of steroid therapy emphasizes the significant role of steroids in the stabilization of the disease.

• 대한본초학회지
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• 제39권1호
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• pp.39-47
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• 2024

Objectives : This study evaluates how various traditional Korean herbal medicines assess MUC5AC expression for esophageal mucosal defense and analyzes the associated mechanisms involved in inflammation. Methods : Forty types of traditional Korean herbal medicines were assessed for in vitro antioxidant activities, and the real-time PCR method was employed to analyze MUC5AC expression under pH 4.5 conditions in human esophageal epithelial cells (HET-1A). Eight types of Korean herbal medicines were evaluated for in vitro antioxidant activities, and Reactive oxygen specise (ROS) expression was analyzed under bile salt (480 𝜇M) and pH 5.5 conditions in human esophageal epithelial cells (HET-1A). Simulation experiments involving bile salts and acidity were conducted for one hour to assess the efficacy of four drugs, and the activities of Mitogen-activated Protein Kinase (MEK), Nuclear Factor Kappa B (NF-𝜅B), and Cyclooxygenase-2 (COX-2) were detected through Western blot analysis. Results : Compared to the Normal group, the Control group exhibited higher ROS generation. Such increased ROS levels were significantly reduced by four extracts: Citrus Unshius Pericarpium (CUP), Cnidium officinale Rhizoma (CR), Ginseng Radix (GR), and Linderae Radix (LR). The protein expression of COX-2 decreased with the treatment of LR, CUP, and CR. Particularly, CUP and CR exhibited superior effects compared to other groups in inhibiting the phosphorylation of NF-𝜅B. Conclusion : Based on the results obtained, we have identified drugs that inhibit oxidative stress and inflammation caused by bile acid in esophageal epithelial cells. Our future plans involve comparing and analyzing the efficacy of these herbal medicines through animal experiments.

• Clinical and Experimental Reproductive Medicine
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• 제30권3호
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• pp.233-240
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• 2003

연구목적: 본 연구에서는 정상 환자와 습관성 유산 질환 환자에서 유래된 융모막 조직 내에서의 면역억제유전자들의 발현 양상에 대해 알아보고자 하였다. 연구재료 및 방법: 임신 6주와 8주의 습관성 유산 질환 환자와 정상 환자로부터 융모막 조직을 채취하였다 (Normal N=6; RSA N=6). 면역조직화학적 분석을 통해서 조직을 관찰하고 세포가 살아있음을 확인한 후에, 역전사 중합효소연쇄반응을 통해서 면역억제유전자인 placenta protein 14 (PP14), indoleamine 2, 3-dioxygenase (IDO) 그리고 mucin1 (MUC1) 유전자의 발현 정도를 비교하였다. GAPDH 발현에 기준한 면역억제유전자 발현을 정량 분석하여 Student's t-test를 시행하였고, p<0.05를 유의성이 있는 것으로 판정하였다. 결 과: 습관성 유산 질환 환자의 경우, 임신 6주와 8주의 융모막 조직에서의 면역억제유전자 (PP14, IDO, MUC1) 발현이 현저하게 낮은 양상을 보이고 있었다. 습관성 유산과 면역억제유전자의 발현이 통계학적으로 유의성 있는 연관성을 가지고 있다는 것이 확인되었다. 결 론: 면역억제유전자 (PP14, IDO, MUC1)의 발현이 습관성 유산 질환 환자에서 특이적으로 낮게 나타나는 것으로 보아 습관성 유산 질환의 진단과 치료 연구 방안에 이 유전자들의 발현이 이용될 수 있을 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권4호
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• pp.489-493
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• 2008

In 2004, Jorgensen and coworkers proposed the MUC4 gene as a candidate gene of enterotoxigenic Escherichia coli (ETEC) F4ab/ac receptor in piglets and a mutation of $G{\rightarrow}C$ in intron 7 of MUC4 was identified to be associated with the ETEC F4ab/ac adhesion phenotypes. In this study, we used 310 piglets of three breeds (Landrace, Large White and Chinese Songliao Black) to analyze the relationship between this mutation and the F4ab/ac adhesion phenotype. The results show that the genotypes at this site and the ETEC F4ab/ac adhesion phenotypes were not completely consistent, although they are very strongly associated. Among the individuals with genotype CC, which was identified as a resistant genotype to F4ab/ac adhesion, only 72.1% (124/172) were non-adhesive to ETEC F4ab and 77.9% (134/172) were non-adhesive to ETEC F4ac infections. This suggests that this mutation may not be the causative mutation for ETEC F4ab/ac adhesion, rather, the actual causative mutation may be in another gene closely linked to MUC4, or at aother site within the MUC4 gene. Our results also suggest that the receptors of F4ab and F4ac may be determined by two different but closely linked loci. In order to screen other genes related to F4ab/ac adhesion in piglets, the mRNA profiles from six full sib piglets, of which three were adhesive to ETEC F4ab/ac and three non-adhesive, were analyzed by suppression subtractive hybridization (SSH). One up-regulated gene, Ep-CAM, was selected for further analysis based on its role in the intestinal epithelial cells adhesion. Using real-time RT-PCR, we found that the Ep-CAM gene was significantly up-regulated in the piglets adhesive to F4ab/ac. It was mapped to SSC3q11-q14 by radiation hybrid mapping.

• 대한한방내과학회지
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• 제31권3호
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• pp.650-666
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• 2010

Objectives : In this study, the author tried to investigate whether wood vinegar produced from Morus alba (MA) significantly affects the increase in airway epithelial mucosubstances and hyperplasia of tracheal goblet cells of rats, and in vitro airway mucin secretion and PMA- or EGF- or TNF-alpha-induced MUC5AC mucin production / gene expression from human airway epithelial cells. Materials and Methods : For the in vivo experiment, the author induced hypersecretion of airway mucus and goblet cell hyperplasia by exposure of rats to SO2 over 3 weeks. Effect of orally-administered MA over 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats and hyperplasia of goblet cells were assessed using histopathological analysis after staining the epithelial tissue with alcian blue. For the in vitro experiment, confluent RTSE cells were chased for 30 min in the presence of MA to assess the effect of MA on mucin secretion by enzyme-linked immunosorbent assay (ELISA). Also, effects of MA on PMA- or EGF- or TNF-alpha-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of MA and treated with PMA (10 ng/ml), EGF (25 ng/ml) or TNF-alpha (0.2 nm) for 24 hrs, to assess both effects of MA on PMA- or EGF- or TNF-alpha-induced MUC5AC mucin production by enzyme-linked immunosorbent assay (ELISA) and gene expression by reverse transcription-polymerase chain reaction (RT-PCR). Possible cytotoxicities of MA in vitro were assessed by examining LDH release from RTSE cells and the rate of survival and proliferation of NCI-H292 cells. In vivo liver and kidney toxicities of MA were evaluated by measuring serum GOT/GPT activities and serum BUN/creatinine concentrations of rats after administering MA orally. Results : 1. MA decreased the amount of intraepithelial mucosubstances of rats exposed to sulfur dioxide inhalationally. 2. MA decreased in vitro mucin secretion from cultured RTSE cells. 3. MA significantly inhibited PMA-, EGF-, and TNF-alpha-induced MUC5AC mucin productions and the expression levels of MUC5AC mRNA from NCI-H292 cells. 4. MA did not show either in vitro or in vivo hepatic or renal toxicities. Conclusion : The results from this study suggests that MA can regulate the secretion, production and gene expression of airway mucin observed in diverse respiratory diseases accompanied by mucus hypersecretion and does not show in vivo toxicity to liver and kidney functions after oral administration. Effects of MA should be further studied using animal experimental models that simulate the diverse pathophysiology of respiratory diseases via future research.