호흡기 상피세포에서 MUC5AC와 MUC5B 발현에 대한 다중벽 탄소나노튜브의 효과

안지훈,김형근,서보현,최윤석,송시연,배창훈,김용대 대한이비인후과학회 2015 대한이비인후과학회지 두경부외과학 Vol.58 No.8

Background and Objectives Multi-walled carbon nanotubes (MWCNT) are one of the most commonly used nanomaterials to date. Recent studies have demonstrated that MWCNT increase immune response and allergic inflammation in airway epithelial cells. However, the effects of MWCNT on mucin in human airway epithelial cells have not been reported. Therefore, in the present study, the effect of MWCNT on MUC16, MUC5AC, and MUC5B expressions were investigated in human airway epithelial cells. Subjects and Method In mucin-producing human NCI-H292 airway epithelial cells and primary cultures of normal nasal epithelial cells, the effects of MWCNT on MUC16, MUC5AC, and MUC5B expression were analyzed by reverse transcription polymerase chain reaction, real-time polymerase chain reaction, and enzyme-linked immunosorbent assay. Results In human NCI-H292 airway epithelial cells, MWCNT significantly induced the expression MUC5AC and MUC5B mRNA and the production of MUC5AC and MUC5B protein. However, MWCNT did not induce the expression of MUC16 mRNA. In the primary cultures of normal nasal epithelial cells, MWCNT also induced the expression of MUC5AC and MUC5B mRNA and the production of MUC5AC and MUC5B proteins. Conclusion The results of this study demonstrate that MWCNT induces MUC5AC and MUC5B expression in human airway epithelial cells. These findings provide important information about the biological role of MWCNT on mucus-secretion in human airway epithelial cells.

사람 호흡기 상피세포에서 MUC5B 발현에 대한 Polyinosinic-Polycytidylic Acid의 효과

최요한,배창훈,김형근,서보현,최윤석,송시연,김용대 대한이비인후과학회 2015 대한이비인후과학회지 두경부외과학 Vol.58 No.9

Background and Objectives Polyinosinic-polycytidylic acid (Poly I:C) is structurally similar to double-stranded RNA, and is known to induce various inflammatory mediators and to cause inflammatory reactions in airway epithelial cells. However, the effect of Poly I:C on secretion of mucins in human airway epithelial cells has been very rarely reported. In this study, the effect and brief signaling pathway of Poly I:C on the expression of mucin genes were investigated in human airway epithelial cells. Materials and Method In mucin-producing human NCI-H292 airway epithelial cells and the primary cultures of normal human nasal epithelial cells, the effect and signaling pathway of Poly I:C on expression of mucin genes were investigated using reverse transcriptase-polymerase chain reaction, real-time PCR, enzyme immunoassay, and immunoblot analysis with specific inhibitors and small interfering RNA (siRNA) for mitogen-activated protein kinase (MAPK). Results Poly I:C induced the MUC5B expression, and activated the phosphorylation of ERK1/2 and p38 MAPK. U0126 (ERK1/2 MAPK inhibitor) and SB203580 (p38 MAPK inhibitor) inhibited the Poly I:C-induced MUC5B expression. In addition, the knockdown of ERK2 and p38 MAPK by siRNA significantly blocked the Poly I:C-induced MUC5B mRNA expression. Conclusion Poly I:C induces the MUC5B expression via ERK2 and p38 MAPK signaling pathways in human airway epithelial cells. Therefore, Poly I:C may play a role in the regulation of mucus hypersecretion through MAPK signaling pathways in the human airway epithelial cells.

인체 호흡기 상피세포에서 담배특이니트로사민에 의한 점액유전자 발현 및 기전

곽소영,최윤석,나형균,배창훈,송시연,김용대 대한비과학회 2020 Journal of rhinology Vol.27 No.1

Background and Objectives: Nicotine is oxidized into tobacco-specific nitrosamines (TSNAs; NAB, NAT, NNN, NNAL, NNK) at high temperature and high pressure. TSNAs are associated with airway diseases characterized by mucus hypersecretion as a major pathophysiologic phenomenon. The aim of study is to investigate the effect of TSNAs on mucin overexpression and its molecular mechanism in human airway epithelial cells. Materials and Method: The cytotoxicity of TSNAs was evaluated using EX-Cytox and inverted microscopy. The mRNA and protein levels of MUC5AC and MUC5B were measured using real-time PCR and ELISA. Results: NAB, NNN, NNAL, and NNK did not affect cell viability. NAT did not affect cell viability up to a concentration of 100 μM in human airway epithelial cells. NAT, NNN, NNAL, and NNK significantly induced MUC5AC expression, but not MUC5B expression. NAB did not affect the expression of MUC5AC and MUC5B. Propranolol (a β-adrenergic receptor antagonist) inhibited NAT, NNN, NNAL, and NNK-induced MUC5AC expression, whereas α-bungarotoxin (an α7-nicotinic acetylcholine receptor antagonist) only inhibited NNN- and NNK-induced MUC5AC expression. Conclusion: These results suggested that NAT, NNN, NNAL, and NNK induce MUC5AC expression through β-adrenergic receptor and/or α7-nicotinic acetylcholine receptor in human airway epithelial cells, which may be involved in mucus hypersecretion in inflammatory airway diseases. Background and Objectives: Nicotine is oxidized into tobacco-specific nitrosamines (TSNAs; NAB, NAT, NNN, NNAL, NNK) at high temperature and high pressure. TSNAs are associated with airway diseases characterized by mucus hypersecretion as a major pathophysiologic phenomenon. The aim of study is to investigate the effect of TSNAs on mucin overexpression and its molecular mechanism in human airway epithelial cells.Materials and Method: The cytotoxicity of TSNAs was evaluated using EX-Cytox and inverted microscopy. The mRNA and protein levels of MUC5AC and MUC5B were measured using real-time PCR and ELISA.Results: NAB, NNN, NNAL, and NNK did not affect cell viability. NAT did not affect cell viability up to a concentration of 100 μM in human airway epithelial cells. NAT, NNN, NNAL, and NNK significantly induced MUC5AC expression, but not MUC5B expression. NAB did not affect the expression of MUC5AC and MUC5B. Propranolol (a β-adrenergic receptor antagonist) inhibited NAT, NNN, NNAL, and NNK-induced MUC5AC expression, whereas α-bungarotoxin (an α7-nicotinic acetylcholine receptor antagonist) only inhibited NNN- and NNK-induced MUC5AC expression.Conclusion: These results suggested that NAT, NNN, NNAL, and NNK induce MUC5AC expression through β-adrenergic receptor and/or α7-nicotinic acetylcholine receptor in human airway epithelial cells, which may be involved in mucus hypersecretion in inflammatory airway diseases.

인간 호흡기 상피세포에서 점액 발현에 대한 Resolvin D1과 E1의 효과

김형근,최태영,배창훈,최윤석,나형균,송시연,김용대 대한이비인후과학회 2019 대한이비인후과학회지 두경부외과학 Vol.62 No.1

Background and Objectives Mucin is an important component of mucus that performs thefirst line of defense against inhaled pathogens and particles, lubrication of organs, and protectionof airway. It is hyper-secreted in inflammatory airway diseases and is associated withmorbidity and mortality of the affected patients. Resolvin, an autacoid of a specific lipid structure,exhibits anti-inflammatory property against inflammatory airway diseases although itseffects on mucin secretion by human airway epithelial cells have not yet been demonstrated. In this regard, we investigated the effects of Resolvin on lipopolysaccharide (LPS)-inducedmucin expression in human airway epithelial cells. Materials and Method In mucin-producing human NCI-H292 epithelial cells, the effectsand brief signaling pathways of Resolvin D1 (RvD1) and Resolvin E1 (RvE1) on the LPS-inducedMUC4, MUC5AC, and MUC5B expression were investigated using reverse transcription-polymerase chain reaction, enzyme-linked immunosorbent assay and Western blot analysis. Results RvD1 attenuated LPS-induced MUC4, MUC5AC, and MUC5B mRNA expressionand protein production in human NCI-H292 cells while RvE1 did not. RvD1 significantlyblocked LPS-induced activated phosphorylation of extracellular signal-regulated kinase 1/2(ERK1/2) mitogen-activated protein kinase (MAPK) and p38 MAPK and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) while RvE1 did not. Conclusion These results suggest that RvD1 attenuates LPS-induced MUC4, MUC5AC,and MUC5B expressions via ERK1/2 MAPK, p38 MAPK, and NF-κB signaling pathways inairway epithelial cells. Therefore, RvD1 may modulate the control of mucus-hypersecretion ininflammatory airway diseases.

사람 호흡기 상피세포에서 Protopanaxadiol의 Lipopolysaccharide로 유도된 활성산소 생성과 MUC5AC 과발현 억제 효과

송유선,김준희,나형균,최윤석,송시연,김용대,배창훈 대한이비인후과학회 2019 대한이비인후과학회지 두경부외과학 Vol.62 No.9

Background and Objectives MUC5AC is one of the major secretory mucin genes in thehuman airway epithelium. MUC5AC expression is increased by a variety of inflammatory mediators. Protopanaxadiol (PPD), one of the major active metabolites in ginseng, is known to haveanti-inflammatory, antitumor and antioxidant properties. However, the effects of PPD on mucinsecretion of airway epithelial cells still have not been reported. Therefore, the aim of this studyis to investigate the effect of PPD on lipopolysaccharide (LPS)-induced MUC5AC expressionin human airway epithelial cells. Materials and Method In the mucin-producing human NCI-H292 airway epithelial cells,the effect of PPD on MUC5AC expression was investigated using reverse transcription-polymerasechain reaction and enzyme immunoassay after treated with LPS. N-acetylcysteine (NAC)as a reactive oxygen species (ROS) scavenger, and apocynin as a nicotinamide adenine dinucleotidephosphate oxidase inhibitor were used to compare the inhibitory effect of PPD on LPSinducedROS production in human NCI-H292 cells. Results LPS significantly increased MUC5AC mRNA expression and protein production. LPS also increased ROS production. PPD inhibited LPS-induced MUC5AC mRNA expressionand protein production as well as ROS production. In addition, NAC and apocynin inhibitedLPS-induced MUC5AC mRNA expression and protein production. Conclusion These results demonstrate that PPD inhibits LPS-induced MUC5AC expressionvia ROS in human airway epithelial cells and the inhibitory effect of PPD was similar to that ofNAC and apocynin. These findings indicate that PPD may be a therapeutic agent for control ofmucus secretion and oxidative stress in human airway epithelial cells.

호흡기 상피세포에서 MUC5AC와 MUC5B 발현에 대한 Betulinic Acid의 효과

김훈성,최윤석,이준혁,박나경,박창휘,이영하,김귀옥,송시연,배창훈,이승호,김용운,김용대 대한이비인후과학회 2014 대한이비인후과학회지 두경부외과학 Vol.57 No.8

Background and Objectives MUC5AC and MUC5B are representative secretory mucin genes in the human airway, whose expressions are increased by a variety of inflammatory mediators. Betulinic acid, a naturally occurring pentacyclic triterpenoid, is known to have an anti-inflammatory property. However, the effects of betulinic acid on mucin secretion of airway epithelial cells still have not been reported. Therefore, in this study, the effect of betulinic acid on inflammatory mediators-induced MUC5AC and MUC5B expressions was investigated in human airway epithelial cells. Materials and Method In the mucin-producing human NCI-H292 airway epithelial cells, the effects of betulinic acid on interleukin-1β (IL-1β)-, lipopolysaccharide (LPS)-, and phorbol myristate acetate (PMA)-induced MUC5AC and MUC5B expressions were analyzed by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. Results Betulinic acid attenuated IL-1β-, LPS-, and PMA-induced MUC5B mRNA and glycoprotein expression in NCI-H292 cells. On the other hand, betulinic acid did not attenuate IL-1β-, and LPS-, but induced PMA-induced MUC5AC mRNA and glycoprotein expressions in NCI-H292 cells. Conclusion These results suggest that betulinic acid attenuates IL-1β-, LPS-, and PMA-induced MUC5B expression in the airway epithelial cells. Therefore, betulinic acid may modulate a control of mucus-hypersecretion in airway inflammatory diseases. Korean J Otorhinolaryngol-Head Neck Surg 2014;57(8):526-32

호흡기 상피세포에서 MUC5AC와 MUC5B 발현에 대한 Betulinic Acid의 효과

김훈성 ( Hoon Sung Kim ),최윤석 ( Yoon Seok Choi ),이준혁 ( Jun Hyeok Lee ),박나경 ( Na Kyung Park ),박창휘 ( Chang Hwi Park ),이영하 ( Young Ha Lee ),김귀옥 ( Gui Ok Kim ),송시연 ( Si Young Song ),배창훈 ( Chang Hoon Bae ),이승호 영남대학교 약품개발연구소 2014 영남대학교 약품개발연구소 연구업적집 Vol.24 No.0

Background and Objectives MUC5AC and MUC5B are representative secretory mucin genes in the human airway. MUC5AC and MUC5B expression are increased by a variety of inflammatory mediators. Betulinic acid, a naturally occurring pentacyclic triterpenoid, is known to have an anti-inflammatory property. However, the effects of betulinic acid on mucin secretion of airway epithelial cells still have not been reported. Therefore, in this study, the effect of betulinic acid on inflammatory mediators-induced MUC5AC and MUC5B expression was investigated in human airway epithelial cells. Materials and Method In the mucin-producing human NCI-H292 airway epithelial cells, the effects of betulinic acid on interleukin-1β (IL-1β)-, lipopolysaccharide (LPS)-, and phorbol myristate acetate (PMA)-induced MUC5AC and MUC5B expression were analyzed by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. Results Betulinic acid attenuated IL-1β-, LPS- and PMA-induced MUC5B mRNA and glycoprotein expression in NCI-H292 cells. Betulinic acid did not attenuate IL-1β-, LPS- and PMA-induced MUC5AC mRNA and glycoprotein expression in NCI-H292 cells. Conclusion These results suggest that betulinic acid attenuates IL-1β-, LPS- and PMA-induced MUC5B expression in airway epithelial cells. Therefore, betulinic acid may modulate a control of mucus-hypersecretion in airway inflammatory disease. Korean J Otorhinolaryngol-Head Neck Surg 2014;57(8):526-32

Delphinidin Inhibits LPS-Induced MUC8 and MUC5B Expression Through Toll-like Receptor 4-Mediated ERK1/2 and p38 MAPK in Human Airway Epithelial Cells

배창훈,전보성,최윤석,송시연,김용대 대한이비인후과학회 2014 Clinical and Experimental Otorhinolaryngology Vol.7 No.3

Objectives. Delphinidin is one of the anthocyanidins. It is believed to have anti-inflammatory property including antioxi- dant, antiangiogenic, and anti-cancer properties. However, the anti-inflammatory effect of delphinidin in mucin-pro- ducing human airway epithelial cells has not been determined. Therefore, this study was conducted in order to investi- gate the effect and the brief signaling pathway of delphinidin in lipopolysaccharide (LPS)-induced MUC8 and MUC5B expression in human airway epithelial cells. Methods. In mucin-producing human NCI-H292 airway epithelial cells and primary cultures of normal nasal epithelial cells, the reverse transcriptase-polymerase chain reaction (RT-PCR), real-time PCR, enzyme immunoassay were used for investigating the expressions of MUC8, MUC5, and Toll-like receptor 4 (TLR4), after LPS treatment and delphin- idin treatment. And the signaling pathway of delphinidin on LPS-induced MUC8 and MUC5B expression was inves- tigated using the RT-PCR, and immunoblot analysis. To confirm the involvement of TLR4 in LPS-induced MUC8 and MU5B expression, the cells were transfected with TLR4 siRNA. Results. In NCI-H292 airway epithelial cells, LPS (100 ng/mL) significantly induced TLR4, MUC8, and MUC5B expres- sion. TLR4 siRNA significantly blocked LPS-induced MUC8 and MUC5B mRNA expression. LPS (100 ng/mL) sig- nificantly activated the phosphorylation of extracellular signal related kinase (ERK) 1/2 and p38 mitogen-activated protein kinase (MAPK). Delphinidin (50 and 100 µM) inhibited LPS-induced TLR4, MUC8, and MUC5B expression and LPS-induced phosphorylation of ERK1/2 and p38 MAPK. In the primary cultures of normal nasal epithelial cells, delphinidin (50 and 100 µM) significantly inhibited LPS-induced TLR4, MUC8, and MUC5B gene expression. Conclusion. These results suggest that delphinidin attenuates LPS-induced MUC8 and MUC5B expression through the TLR4-mediated ERK1/2 and p38 MAPK signaling pathway in human airway epithelial cells. These findings indicated that delphinidin may be a therapeutic agent for control of inflammatory airway diseases.

인체 호흡기 상피세포에서 고농도 인슐린에 의한 MUC4, MUC5AC, MUC5B 점액 유전자의 발현

나형균,배창훈,최윤석,송시연,진현정,김용대 대한비과학회 2016 Journal of rhinology Vol.23 No.1

Background and Objectives: Insulin is a peptide hormone that regulates the metabolism of carbohydrates and fats by promoting the absorption of glucose from the blood to skeletal muscles. Insulin has been reported to be closely related to cardiovascular, respiratory, and endocrine disease. However, the effect of insulin on production of major mucins in human airway epithelial cells has not been reported. Therefore, this study investigated the relationship between high levels of insulin and mucin in human airway epithelial cells. Materials and Methods: This study analyzed the effect of high level of insulin on MUC4, MUC5AC, and MUC5B expression using reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay in human airway epithelial cells. Results: In human NCI-H292 airway epithelial cells, high level of insulin significant increased MUC4, MUC5AC, and MUC5B mRNA expression and glycoprotein production. In the primary cultures of normal nasal epithelial cells, high level of insulin also increased MUC4, MUC5AC, and MUC5B expression. Conclusion: These results suggest that insulin plays a role in control of mucus hypersecretion in human airway epithelial cells.

사람 호흡기 상피세포에서 Triptolide의 Nuclear Factor-Kappa B를 통한 Lipopolysaccharide로 유도된 MUC5AC/5B 발현 억제 효과

서보현,최태영,최윤석,배창훈,나형균,송시연,김용대 대한이비인후과학회 2018 대한이비인후과학회지 두경부외과학 Vol.61 No.12

Background and Objectives The representative mucin genes in the human airway areMUC5AC and MUC5B, which are regulated by several inflammatory and anti-inflammatorysubstances. Triptolide (TPL), udenafil, betulinic acid, changkil saponin, and glucosteroid aresome of the many anti-inflammatory substances that exist. TPL is a diterpenoid compound fromthe thunder god vine, which is used in traditional Chinese medicine for treatment of immuneinflammatory diseases, such as rheumatoid arthritis, systemic lupus erythematosus, nephritisand asthma. However, the effects of TPL on mucin expression of human airway epithelial cellshave yet to be reported. Hence, this study investigated the effect of TPL on lipopolysaccharide(LPS)-induced MUC5AC and MUC5B expression in human airway epithelial cells. Subjects and Method The NCI-H292 cells and the primary cultures of human nasal epithelialcells were used to investigate the effects of TPL on LPS-induced MUC5AC and MUC5B expressionusing real-time polymerase chain reaction, enzyme immunoassay, and Western blot. Results TPL significantly decreased the LPS-induced MUC5AC and MUC5B mRNA expressionand protein production. TPL also significantly decreased the nuclear factor-kappa B(NF-kB) phosphorylation. Conclusion These results suggest that TPL down regulates MUC5AC and MUC5B expressionvia inhibition of NF-kB activation in human airway epithelial cells. This study may provideimportant information about the biological role of triptolide on mucus-secretion in airwayinflammatory diseases and the development of novel therapeutic agents for controlling suchdiseases.