• Molecules and Cells
• /
• 제46권11호
• /
• pp.700-709
• /
• 2023

Mucus hyperproduction and hypersecretion are observed often in respiratory diseases. MUC8 is a glycoprotein synthesized by epithelial cells and generally expressed in the respiratory track. However, the physiological mechanism by which extracellular nucleotides induce MUC8 gene expression in human airway epithelial cells is unclear. Here, we show that UTP could induce MUC8 gene expression through P2Y2-PLCβ3-Ca²⁺ activation. Because the full-length cDNA sequence of MUC8 has not been identified, a specific siRNA-MUC8 was designed based on the partial cDNA sequence of MUC8. siRNA-MUC8 significantly increased TNF-α production and decreased IL-1Ra production, suggesting that MUC8 may downregulate UTP/P2Y2-induced airway inflammation. Interestingly, the PDZ peptide of ZO-1 protein strongly abolished UTP-induced TNF-α production and increased IL-1Ra production and MUC8 gene expression. In addition, the PDZ peptide dramatically increased the levels of UTP-induced ZO proteins and TEER (trans-epithelial electrical resistance). These results show that the anti-inflammatory mucin MUC8 may contribute to homeostasis, and the PDZ peptide can be a novel therapeutic candidate for UTP-induced airway inflammation.

• 대한치과교정학회지
• /
• 제32권6호
• /
• pp.443-453
• /
• 2002

본 연구의 목적은 다양한 교정용 브라켓의 표면에 형성되는 타액성 피막의 조성을 확인하고, 전타액, 악하선타액 및 이 하선타액에서 유래하는 타액성 피막의 성분을 비교하는 것이다. 네 가지 서로 다른 종류의 교정용 브라켓을 본 연구에 사용하였다. 이들은 $022{\times}028$ Roth Prescription의 상악 소구치 브라켓으로 조성은 다음과 같다: 스테인레스 스틸, 단결정 사파이어, 다결정 알루미나 및 플라스틱 브라켓. 교정용 브라켓을 각각 전타액, 이하선타액 및 악하선타액에 2시간 배양하여 타액성 피막을 형성시켰다 브라켓 피막의 타액성분은 sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western transfer method 및 면역검출법을 통해 확인하였다. 이 결과 low-molecular weight salivary mucin, ${\alpha}-amylase$, secretory IgA (sIgA), acidic proline-rich proteins, cystatins 등이 모든 브라켓의 타액성 피막에 존재하였으며, 치아우식증의 원인균인 Streptococcus mutans의 부착을 촉진시키는 타액단백질인 high-molecular weight mucin은 어떤 브라켓에도 부착하지 않았다. 그러나, 비록 동일한 타액단백질이 모든 브라켓에서 발견되었지만, 타액단백질 부착 양상은 타액의 종류 및 브라켓의 종류에 따라 양적 및 질적으로 다르게 나타났다. 특히 sIgA는 이하선타액에서 유래한 브라켓 피막에 더 많이 부착하였고, cystatins의 경우는 플라스틱 브라켓에서 유래한 브라켓 피막에 더 많이 존재하였다 본 연구는 다양한 타액단백질이 교정용 브라켓에 부착하며, 타액단백질이 타액의 출처 및 브라켓의 종류에 따라 교정용 브라켓의 표면에 선택적으로 부착함을 나타내었다.

• Tuberculosis and Respiratory Diseases
• /
• 제64권5호
• /
• pp.347-355
• /
• 2008

연구배경: 특발성폐섬유증 환자의 전형적인 증상은 운동호흡곤란과 마른기침으로, 객담이 적은 원인에 대해서 아직까지 잘 알려져 있지 않다. Interferon-${\gamma}$ inducible protein-10 (IP-10)은 여러 호흡기질환과 관련되는데 폐 내로 염증을 유입시키는데 중요한 역할을 한다. 본 연구는 특발성폐섬유증 환자에서 객담이 적은 기전으로 IP-10이 연관성이 있는지에 대해 연구하였다. 방법: 특발성폐섬유증 환자를 대상으로 기관지폐포세척액에서 IP-10의 농도를 ELISA로 측정하였다. IP-10이 기도 점액소 발현에 미치는 영향을 간접적으로 알아보기 위해 NCI-H292 세포(점막표피모양 암종 세포주)에서 IP-10을 전처치한 이후 tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)로 자극하여 발현정도를 측정하였다. 이때 점액소 발현과 관련되는 기전으로 epidermal growth factor receptor-mitogen activated protein kinase (EGFR-MAPK)의 신호전달 경로를 알아 보았다. 결과: IP-10의 기관지폐포세척액내 농도는 특발성폐섬유증 환자가 건강 대조군에 비해 유의하게 높았다. IP-10의 전처치는 NCI-H292 세포에서 TNF-${\alpha}$ 유도 MUC5AC 점액소 발현을 감소시켰는데 이 때 EGFR-MAPK 신호전달 경로의 차단과 관련되었다. 결론: 특발성폐섬유증 환자의 적은 객담은 IP-10의 발현증가와 일부 관련 가능성이 있으며, 이때 IP-10의 작용은 MUC5AC 점액소 유전자 발현에 필요한 EGFR-MAPK 신호전달 경로의 차단과 관련될 것으로 생각한다.

• BMB Reports
• /
• 제45권11호
• /
• pp.665-670
• /
• 2012

The insect midgut epithelium is generally lined with a unique chitin and protein structure, the peritrophic membrane (PM), which facilitates food digestion and protects the gut epithelium. We used gel electrophoresis and mass spectrometry to identify the extracted proteins from the silkworm PM to obtain an in-depth understanding of the biological function of the silkworm PM components. A total of 305 proteins, with molecular weights ranging from 8.02 kDa to 788.52 kDa and the isoelectric points ranging from 3.39 to 12.91, were successfully identified. We also found several major classes of PM proteins, i.e. PM chitin-binding protein, invertebrate intestinal mucin, and chitin deacetylase. The protein profile provides a basis for further study of the physiological events in the PM of Bombyx mori.

• IMMUNE NETWORK
• /
• 제9권2호
• /
• pp.58-63
• /
• 2009

Background: T cell immunoglobulin and mucin domain containing 3 protein (Tim-3) expressed on terminally differentiated Th1 cells plays a suppressive role in Th1-mediated immune responses. Recently, it has been shown that N-glycosylation affects the binding activity of the Tim-3-Ig fusion protein to its ligand, galectin-9, but the binding properties of non-glycosylated Tim-3 on $CD4^+CD25^+$T cells has not been fully examined. In this study, we produced recombinant Tim-3-Ig fusion proteins in different cellular sources and its N-glycosylation mutant forms to evaluate their binding activities to $CD4^+CD25^+$T cells. Methods: We isolated and cloned Tim-3 cDNA from BALB/C mouse splenocytes. Then, we constructed a mammalian expression vector and a prokaryotic expression vector for the Tim-3-Ig fusion protein. Using a site directed mutagenesis method, plasmid vectors for Tim-3-Ig N-glycosylation mutant expression were produced. The recombinant protein was purified by protein A sepharose column chromatography. The binding activity of Tim-3-Ig fusion protein to $CD4^+CD25^+$T cells was analyzed using flow cytometry. Results: We found that the nonglycosylated Tim-3-Ig fusion proteins expressed in bacteria bound to $CD4^+CD25^+$T cells similarly to the glycosylated Tim-3-Ig protein produced in CHO cells. Further, three N-glycosylation mutant forms (N53Q, N100Q, N53/100Q) of Tim-3-Ig showed similar binding activities to those of wild type glycosylated Tim-3-Ig. Conclusion: Our results suggest that N-glycosylation of Tim-3 may not affect its binding activity to ligands expressed on $CD4^+CD25^+$T cells.

• 약학회지
• /
• 제50권3호
• /
• pp.166-171
• /
• 2006

A calcium-dependent sialic acid-binding lectin has been isolated by thyroglobulin-affinity chromatography from the coastal crab Macrophthalmus Japonicus. This lectin, Macrophthalmus Japonicus lectin (MJL), was eluted with 50mM Tris-HCl, 0.3 M NaCl, 10 mM EDTA, and the recovery yield from the crude protein extract was about 5.6%. The molecular weight of MJL was estimated as 65 kDa in SDS-PAGE both under reducing and non-reducing conditions. MJL induced an agglutination reaction in rabbit, rat, and mouse erythrocytes, but not in human ABO types. This activity was effectively inhibited by sialoglycoproteins such as fetuin, bovine submaxillary mucin, and thyroglobulin.

• Biomolecules & Therapeutics
• /
• 제32권4호
• /
• pp.451-459
• /
• 2024

Apoptosis signal-regulating kinase 1 (ASK1) is an upstream signaling molecule in oxidative stress-induced responses. Because oxidative stress is involved in asthma pathogenesis, ASK1 gene deficiency was investigated in animal models of allergic asthma. However, there is no study to investigate whether ASK1 inhibitors could be applied for asthma to date. Selonsertib, a potent and selective ASK1 inhibitor, was applied to BALB/c mice of an ovalbumin (OVA)-induced allergic asthma model. Selonsertib suppressed antigen-induced degranulation of RBL-2H3 mast cells in a concentration-dependent manner. The administration of selonsertib both before OVA sensitization and OVA challenge significantly reduced airway hyperresponsiveness, and suppressed eosinophil numbers and inflammatory cytokine levels in the bronchoalveolar lavage fluid. Histopathologic examination elucidated less inflammatory responses and reduced mucin-producing cells around the peribronchial regions of the lungs. Selonsertib also suppressed the IgE levels in serum and the protein levels of IL-13 in the bronchoalveolar lavage fluid. These results suggest that selonsertib may ameliorate allergic asthma by suppressing immune responses and be applicable to allergic asthma.

• Asian Pacific Journal of Cancer Prevention
• /
• 제13권10호
• /
• pp.5257-5261
• /
• 2012

Background: High incidence of breast cancer and its fatal effect has reached an alarming stage across the globe, including the third world countries. Many factors have been reported to be associated with the development of breast cancer but detailed structural and functional information is missing. CA 15-3 is one of the known potential tumor marker of breast cancer; however little is known about structure and functional site of this protein. Present study aims to investigate the functional role of CA 15-3 in breast cancer, especially in development and metastasis. Material and Methods: Hundred female breast cancer patients confirmed by histopathological reports were included in the study. Their physiological characters were recorded in a performa. Enzyme linked immunosorbent assay (ELISA) technique was used to estimate serum CA 15-3 level. Immunohistochemistry was done for estrogen (ER), progesterone (PR) and Her2/neu receptors expression. Results: The study revealed the details of physiological characteristics of female breast cancer. Mean age was $37.72{\pm}5.99$ and $55.05{\pm}7.28$ years and serum CA 15-3 (MUC1) level was $60.47{\pm}8.59$ and $63.17{\pm}4.58$ U/ml in pre and post-menopause respectively, and both groups of women had sedentary life style. Their receptor status especially of progesterone, estrogen and HER-2/neu were positive in 50% of premenopausal women and 65% of postmenopausal women. Conclusion: There are multiple physiological factors promoting breast cancer. High serum CA 15-3 level and hormonal imbalance of ER, PR and Her2/neu appears to be the main cause of breast cancer. It may be possible that the functional sites of these proteins may be altered which may increase the chances of metastasis in breast cancer.

• Asian Pacific Journal of Cancer Prevention
• /
• 제13권4호
• /
• pp.1569-1573
• /
• 2012

Luteolin (LUT), a bioflavonoid has been used as a chemopreventive agent world-wide against chemically induced cancer. Hence we designed an experiment to assess chemopreventive action of LUT on lipid peroxidation (LPO) and glycoconjugates in azoxymethane (AOM)-induced colon carcinogenesis. Colon cancer was induced by 15 mg/body kg. body weight of AOM and administration of LUT (at the dose of 1.2 mg/kg. body weight) was till end of the study. Analysis of lipid peroxidative end products such as protein carbonyl (PC), malonadehyde (MDA) and conjucated dienes (CD) demonstrated significant increase in in AOM-induced animals with reduction by LUT (p<0.05). Increased levels of glycoconjugates such as hexose, hexosamine, sialic acid, fucose and mucoprotein were analyzed in serum and colon tissues examined histopathologically by periodic acid Schiff's (PAS) staining were also reversed by LUT l(p<0.05). The secondary marker of colon cancer mucin depleted foci (MDF) was assessed in control and experimental group of animals. A characteristic increase of MDF was observed in AOM-induced colon cancer animals. Treatment with LUT decreased the incidence of MDF. These results suggest that LUT alters the expression of glycoconjugates and suppress colon cancer. Hence, we speculate that LUT can be used as a chemopreventive agent to treat colon cancer.

• Biomolecules & Therapeutics
• /
• 제29권4호
• /
• pp.427-433
• /
• 2021

Free fatty acid receptor 2 (FFA2, also known as GPR43), a G-protein-coupled receptor, has been known to recognize short-chain fatty acids and regulate inflammatory responses. FFA2 gene deficiency exacerbated disease states in several models of inflammatory conditions including asthma. However, in vivo efficacy of FFA2 agonists has not been tested in allergic asthma. Thus, we investigated effect of 4-chloro-α-(1-methylethyl)-N-2-thiazoylylbenzeneacetanilide (4-CMTB), a FFA2 agonist, on antigen-induced degranulation in RBL-2H3 cells and ovalbumin-induced allergic asthma in BALB/c mice. Treatment of 4-CMTB inhibited the antigen-induced degranulation concentration-dependently. Administration of 4-CMTB decreased the immune cell numbers in the bronchoalveolar lavage fluid and suppressed the expression of inflammatory Th2 cytokines (IL-4, IL-5, and IL-13) in the lung tissues. Histological studies revealed that 4-CMTB suppressed mucin production and inflammation in the lungs. Thus, results proved that FFA2 functions to suppress allergic asthma, suggesting 4-CMTB activation of FFA2 as a therapeutic tool for allergic asthma.