• BMB Reports
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• 제29권3호
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• pp.248-252
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• 1996

The epidermal organ of the leech contains a complex glycoprotein molecule of mucus. The mucus excreted from annelids plays Significant role in protection against desiccation and parasites. Mucus from the Korean native leech, H. nipponia, was investigated for biochemical characteristics for possible development of biomaterials of cosmetic and pharmaceutical agents. The leech skin mucus was heavily glycosylated mucin-like protein with a high molecular weight comprised 80% carbohydrate and 20% protein. Threonine, serine, and glycine were the major components of the isolated protein and these accounted for 50% of total amino acids. The carbohydrate portion contained glucosamine, galactosamine. galactose, glucose. mannose and sialic acid in oligosaccharide form linked with threonine and serine residues of the glycoprotein.

• Parasites, Hosts and Diseases
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• 제54권6호
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• pp.751-758
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• 2016

This study aimed at constructing a draft genome of the adult female worm Toxocara canis using next-generation sequencing (NGS) and de novo assembly, as well as to find new genes after annotation using functional genomics tools. Using an NGS machine, we produced DNA read data of T. canis. The de novo assembly of the read data was performed using SOAPdenovo. RNA read data were assembled using Trinity. Structural annotation, homology search, functional annotation, classification of protein domains, and KEGG pathway analysis were carried out. Besides them, recently developed tools such as MAKER, PASA, Evidence Modeler, and Blast2GO were used. The scaffold DNA was obtained, the N50 was 108,950 bp, and the overall length was 341,776,187 bp. The N50 of the transcriptome was 940 bp, and its length was 53,046,952 bp. The GC content of the entire genome was 39.3%. The total number of genes was 20,178, and the total number of protein sequences was 22,358. Of the 22,358 protein sequences, 4,992 were newly observed in T. canis. Following proteins previously unknown were found: E3 ubiquitin-protein ligase cbl-b and antigen T-cell receptor, zeta chain for T-cell and B-cell regulation; endoprotease bli-4 for cuticle metabolism; mucin 12Ea and polymorphic mucin variant C6/1/40r2.1 for mucin production; tropomodulin-family protein and ryanodine receptor calcium release channels for muscle movement. We were able to find new hypothetical polypeptides sequences unique to T. canis, and the findings of this study are capable of serving as a basis for extending our biological understanding of T. canis.

• Tuberculosis and Respiratory Diseases
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• 제56권3호
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• pp.289-296
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• 2004

연구배경 : 기도 질환에서 점액이 과량 분비되는 경우 환자에게 불편함을 줄뿐만 아니라 기도 질환 예후에도 나쁜 영향을 미친다. 그러나, 기도 질환에서 점액이 과량 분비되는 것을 효과적으로 막는 방법이 없다. 점액의 성분 중 mucin은 당화 단백질로서 점액이 점성을 띄게 만드는 주요 성분이다. 본 연구를 통해서 mucin 분비 기전에 proteinase가 관여하는지 확인하고 만일 proteinase가 mucin 분비기전에 관여 한다면 어느 proteinase가 그런 역할을 하는지 확인하고자 하였다. 방 법 : (1) mucin 분비 억제 실험 군 특이적 proteinase 억제제를 사용하여 어느 군에 속하는 proteinase가 mucin 분비를 억제하는지 mucin을 생산하는 폐 세포주인 Calu-3를 이용하여 알아보았다. 군 특이적 proteinase 억제제로 PMSF(phenylmethylsulfonyl fluoride, serine proteinase inhibitor), E-64(cysteine proteinase inhibitor), Pepstatin(aspartic proteinase inhibitor), 1,10-Phenanthroline(metalloproteinase inhibitor)를사용하였다. 군 특이적 억제제를 Calu-3에 24시간동안 처리하여 분비된 mucin양을 enzyme linked immunoabsorbant assay(MUC5AC)로 정량하였고 그 결과를 대조군과 비교하였다. (2) Mucin 분비 자극 실험 Metalloproteinase 중에서 기도 질환 발병과 관련 있다고 알려진 matrix metalloproteinase-9 (MMP-9), MMP-12 그리고 TNF-alpha converting enzyme(TACE)를 Calu-3에 24시간 처리하여 분비된 mucin양을 enzyme linked immunoabsorbant assay (MUC5AC)로 정량하였고 그 결과를 대조군과 비교하였다. 결 과 : (1) 군 특이적 proteinase 억제제인 PMSF($10^{-4}M$), E-64($10^{-4}M$), Pepstatin($10^{-6}M$), 1,10-Phenanthroline($10^{-4}M$)는 MUC5AC 분비를 각각 $1{\pm}4.9%$(평균${\pm}$표준오차; 대조군과 비교 시 P=1.0), $-6{\pm}3.9%$ (P=0.34), $-13{\pm}9.7%$(P=0.34), $41{\pm}8.2%$(P=0.03) 감소시켰다(실험 회수 4번). (2) MMP-9(250ng/ml), MMP-12(100ng/ml), TACE(200ng/ml)에 의한 MUC5AC 분비량은 대조군에 비하여 각각 $103{\pm}6%$(P=0.39), $102{\pm}8%$(P=1.0), $107{\pm}13%$(P=0.39)이었다(실험 회수 6번). 결 론 : mucin 분비 기전에 metalloproteinase가 관여함을 시사하지만 MMP-9, MMP-12, TACE는 in vitro 모델에서 mucin 분비에 영향을 미치지 않았다.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1995년도 한국생물과학협회 학술발표대회
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• pp.298.2-298
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• 1995

No Abstract, See Full Text

• Natural Product Sciences
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• 제25권2호
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• pp.103-110
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• 2019

We investigated the anti-inflammatory effect of Pyunkang-tang extract (PGT), a complex herbal extract based on traditional Chinese medicine that is used in Korea for controlling diverse pulmonary diseases, on cigarette smoke-induced pulmonary pathology in a rat model of chronic obstructive pulmonary disease (COPD). The constituents of PGT were Lonicerae japonica, Liriope platyphylla, Adenophora triphilla, Xantium strumarinum, Selaginella tamariscina and Rehmannia glutinosa. Rats were exposed by inhalation to a mixture of cigarette smoke extract (CSE) and sulfur dioxide for three weeks to induce COPD-like pulmonary inflammation. PGT was administered orally to rats and pathological changes to the pulmonary system were examined in each group of animals through measurement of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and interleukin-6 (IL-6) levels in bronchoalveolar lavage fluid (BALF) at 21 days post-CSE treatment. The effect of PGT on the hypersecretion of pulmonary mucin in rats was assessed by quantification of the amount of mucus secreted and by examining histopathologic changes in tracheal epithelium. Confluent NCI-H292 cells were pretreated with PGT for 30 min and then stimulated with CSE plus PMA (phorbol 12-myristate 13-acetate), for 24 h. The MUC5AC mucin gene expression was measured by RT-PCR. Production of MUC5AC mucin protein was measured by ELISA. The results were as follows: (1) PGT inhibited CSE-induced pulmonary inflammation as shown by decreased TNF-${\alpha}$ and IL-6 levels in BALF; (2) PGT inhibited the hypersecretion of pulmonary mucin and normalized the increased amount of mucosubstances in goblet cells of the CSE-induced COPD rat model; (3) PGT inhibited CSE-induced MUC5AC mucin production and gene expression in vitro in NCI-H292 cells, a human airway epithelial cell line. These results suggest that PGT might regulate the inflammatory aspects of COPD in a rat model.

• Journal of Microbiology and Biotechnology
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• 제27권1호
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• pp.49-56
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• 2017

In the human airway, mucus exists to protect the respiratory system as a primary barrier of the innate immune system. However, hyperexpressed mucus limits airflow, resulting in a decrease of lung function. Among more than 20 mucin family members, MUC5AC and MUC5B are major glycoproteins in human airway mucus. The epidermal growth factor receptor (EGFR) signaling pathway is one of the mechanisms of these mucins expression and specificity protein-1 (Sp1) transcription factor is the downstream signal of this pathway, playing pivotal roles in mucin expression. Even though there are some drugs for treating mucus hypersecretion, no drug has proven effects on humans. We found that the flavonoid tilianin regulated MUC5AC expression and also inhibited Sp1 phosphorylation. In this study, we investigated how tilianin would modulate EGFR signaling and regulate mucin production. In conclusion, tilianin inhibited MUC5AC expression mediated via modulating the EGFR-MEK-ERK-Sp1 signaling pathway in NCI-H292 human airway epithelial cells. This study may provide the basis for the novel treatment of mucus hypersecretion.

• Tuberculosis and Respiratory Diseases
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• 제75권5호
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• pp.205-209
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• 2013

Background: We investigated whether prunetin significantly affects tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-induced MUC5AC mucin gene expression, production, inhibitory kappa B ($I{\kappa}B$) degradation and nuclear factor kappa B (NF-kB) p65 translocation in human airway epithelial cells. Methods: Confluent NCI-H292 cells were pretreated with prunetin for 30 minutes and then stimulated with TNF-${\alpha}$ for 24 hours or the indicated periods. MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The effect of prunetin on TNF-${\alpha}$-induced degradation of $I{\kappa}B$ and translocation of NF-${\kappa}B$ p65 was investigated by western blot analysis. Results: We found that incubation of NCI-H292 cells with prunetin significantly inhibited mucin production and down-regulated the MUC5AC gene expression induced by TNF-${\alpha}$. Prunetin inhibited TNF-${\alpha}$-induced degradation of $I{\kappa}B$ and translocation of NF-${\kappa}B$ p65. Conclusion: This result suggests that prunetin inhibits the NF-${\kappa}B$ signaling pathway, which may explain its role in the inhibition of MUC5AC mucin gene expression and production regulated by the NF-${\kappa}B$ signaling pathway.

• Journal of Ginseng Research
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• 제47권1호
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• pp.97-105
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• 2023

Background: Hyperactivated airway mucosa cells overproduce mucin and cause severe breathing complications. Here, we aimed to identify the effects of saponins derived from Panax ginseng on inflammation and mucin overproduction. Methods: NCI-H292 cells were pre-incubated with 16 saponins derived from P. ginseng, and mucin overproduction was induced by treatment with phorbol 12-myristate 13-acetate (PMA). Mucin protein MUC5AC was quantified by enzyme-linked immunosorbent assay, and mRNA levels were analyzed using quantitative polymerase chain reaction (qPCR). Moreover, we performed a transcriptome analysis of PMA-treated NCI-H292 cells in the absence or presence of Rg5, and differential gene expression was confirmed using qPCR. Phosphorylation levels of signaling molecules, and the abundance of lipid droplets, were measured by western blotting, flow cytometry, and confocal microscopy. Results: Ginsenoside Rg5 effectively reduced MUC5AC secretion and decreased MUC5AC mRNA levels. A systematic functional network analysis revealed that Rg5 upregulated cholesterol and glycerolipid metabolism, resulting in the production of lipid droplets to clear reactive oxygen species (ROS), and modulated the mitogen-activated protein kinase and nuclear factor (NF)-kB signaling pathways to regulate inflammatory responses. Rg5 induced the accumulation of lipid droplets and decreased cellular ROS levels, and N-acetyl-ⳑ-cysteine, a ROS inhibitor, reduced MUC5AC secretion via Rg5. Furthermore, Rg5 hampered the phosphorylation of extracellular signal-regulated kinase and p38 proteins, affecting the NF-kB signaling pathway and pro-inflammatory responses. Conclusion: Rg5 alleviated inflammatory responses by reducing mucin secretion and promoting lipid droplet-mediated ROS clearance. Therefore, Rg5 may have potential as a therapeutic agent to alleviate respiratory disorders caused by hyperactivation of mucosa cells.

• Tuberculosis and Respiratory Diseases
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• 제55권1호
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• pp.21-30
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• 2003

서 론 : MUC genes의 증가와 배상세포의 증식 기전에 성장인자(growth factor)인 상피세포 성장인자 및 수용체(epidermal growth factor receptor; EGFR)가 배상세포의 증식이나 이형성에 관여한다. EGFR의 ligands 중의 한 종류인 heparin binding EGF(HB-EGF)는 세포막에 존재하는 pro-heparin binding EGF(pro-HB-EGF)로부터 유리된다. HB-EGF의 유리는 G-protein과 연관이 있다. 따라서, 본 연구는 그람 음성세균의 lipopolysaccande(LPS)에 의한 기도 점액 과생성의 기전을 밝히고, 기도점액 과분비에서 EGFR과 G-protein의 연관성을 밝혀 기도 점액 과분비 기전을 밝히고자 한다. 연구방법 : NCI-H292 세포배양에서 LPS단독 투여 또는TGF-${\alpha}$와 병합 투여한 후 MUC5AC의 당단백질을 ELISA법으로 측정하였다. LPS에 의한 MUC5AC 당단백질의 생성 기전을 밝히기 위해서 heterotrimeric G-protein 억제제인 mastoparan을 투여하고 TNF-${\alpha}$와 MUC5AC를 ELISA법으로 각각 측정하였다. MUC5AC의 생성에서 G-protein과 EGFR의 연관성을 확인하기 위하여 EGFR이 항상 발현되어 있고 MUC5AC를 분비할 수 있는 NCI-H292 세포에 G-protein 자극제인 mastoparan-7로 자극한 후 MUC5AC의 생성을 측정하였다. G-protein이 활성화하여 metalloproteinase가 세포막에 있는 HB-EGF를 유리하여 EGFR이 활성화하여 MUC5AC가 생성여부를 확인하기 위하여 ADAM10으로 NCI-H292세포에 자극하여 MUC5AC의 생성을 측정하였다. MUC5AC 생성이 EGFR과 연관성을 확인하기 위하여 특이 EGFR tyrosine kinase 억제제인 AG1478과 중화 polyclonal EGF 항체를 전처치 후 MUC5AC를 측정하였다. 결 과 : LPS의 자극에 의한 MUC5AC의 생성은 LPS 농도에 유의하게 증가 되지 않았으나, EGFR의 ligand인 TGF-${\alpha}$를 동시 투여한 경우는 LPS의 농도에 비례하여 유의하게 증가하였다. LPS의 자극은 TNF-${\alpha}$의 생성을 유의하게 증가시켰으며, G-protein 억제제인 mastoparan을 전처치한 경우는 TNF-${\alpha}$가 유의하게 감소 되었다. LPS 자극 전에 TNF-${\alpha}$ antibody, AG1478 또는 mastoparan을 전처치한 경우는 MUC5AC의 생성이 유의하게 억제되었다. MUC5AC의 생생에서 G-protein과 EGFR의 연관성에 대한 실험에서 MUC5AC의 생성이 mastoparan-7의 농도에 따라 유의하게 증가되었으며, EGF의 중화항체를 사용한 경우는 MUC5AC의 생성이 감소되었다. 또한 Matrix metalloproteinase인 ADAM10의 농도에 비례하여 MUC5AC의 생성을 증가시켰다. 결 론 : LPS에 의한 MUC5AC의 분비는 LPS가 TNF-${\alpha}$를 생성시키고, TNF-${\alpha}$가 EGFR의 발현을 유도하여 MUC5AC가 분비되었다. 또한 MUC5AC의 생성에 있어서 G-protein의 활성은 matrix metalloproteinase에 의하여 EGFR의 ligand 인 HB-EGF가 유리되어 EGFR의 transacti vation으로 MUC5AC가 생성되는 것으로 사료된다.

• Genomics & Informatics
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• 제21권4호
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• pp.46.1-46.10
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• 2023

Colon adenocarcinoma (COAD) is the predominant type of colorectal cancer. Early diagnosis and treatment can significantly improve the prognosis of COAD patients. Anoctamin 7 (ANO7), an anion channel protein, has been implicated in prostate cancer and other types of cancer. In this study, we analyzed the expression of ANO7 and its correlation with clinicopathological characteristics among COAD patients using the Gene Expression Profiling Interactive Analysis 2 (GEPIA2) and the University of Alabama at Birmingham CANcer (UALCAN) databases. The GEPIA2, Kaplan-Meier plotter, and the Survival Genie platform were employed for survival analysis. The co-expression network and potential function of ANO7 in COAD were analyzed using GeneFriends, the Database for Annotation, Visualization and Integrated Discovery (DAVID), GeneMANIA, and Pathway Studio. Our data analysis revealed a significant reduction in ANO7 expression levels within COAD tissues compared to normal tissues. Additionally, ANO7 expression was found to be associated with race and histological subtype. The COAD patients exhibiting low ANO7 expression had lower survival rates compared to those with high ANO7 expression. The genes correlated with ANO7 were significantly enriched in proteolysis and mucin type O-glycan biosynthesis pathway. Furthermore, ANO7 demonstrated a direct interaction and a positive co-expression correlation with mucin 2 (MUC2). In conclusion, our findings suggest that ANO7 might serve as a potential prognostic biomarker and potentially plays a role in proteolysis and mucin biosynthesis in the context of COAD.