MUC Expression in Gallbladder Epithelial Tissues in Cholesterol-Associated Gallbladder Disease

( Kyo-sang Yoo ),( Ho Soon Choi ),( Dae Won Jun ),( Hang Lak Lee ),( Oh Young Lee ),( Byung Chul Yoon ),( Kyeong Geun Lee ),( Seung Sam Paik ),( Yong Seok Kim ),( Jin Lee ) 대한간학회 2016 Gut and Liver Vol.10 No.5

Background/Aims: Gallstone pathogenesis is linked to mucin hypersecretion and bacterial infection. Several mucin genes have been identified in gallbladder epithelial cells (GBECs). We investigated MUC expression in cholesterolassociated gallbladder disease and evaluated the relationship between mucin and bacterial infection. Methods: The present study involved 20 patients with cholesterol stones with cholecystitis, five with cholesterol stones with cholesterolosis, six with cholesterol polyps, two with gallbladder cancer, and six controls. Canine GBECs treated with lipopolysaccharide were also studied. MUC3, MUC5AC, MUC5B, and MUC6 antibodies were used for dot/slot immunoblotting and immunohistochemical studies of the gallbladder epithelial tissues, canine GBECs, and bile. Reverse-transcription polymerase chain reaction was performed to evaluate MUC3 and MUC5B expression. Results: MUC3, MUC5AC, MUC5B, and MUC6 were expressed in the normal gallbladder epithelium, and of those, MUC3 and MUC5B exhibited the highest expression levels. Greatly increased levels of MUC3 and MUC5B expression were observed in the cholesterol stone group, and slightly increased levels were observed in the cholesterol polyp group; MUC3 and MUC5B mRNA was also upregulated in those groups. Canine GBECs treated with lipopolysaccharide also showed upregulation of MUC3 and MUC5B. Conclusions: The mucin genes with the highest expression levels in gallbladder tissue in cholesterol-associated diseases were MUC3 and MUC5B. Cholesterol stones and gallbladder infections were associated with increased MUC3 and MUC5B expression. (Gut Liver 2016;10:851-858)

호흡기 상피세포에서 MUC5AC와 MUC5B 발현에 대한 Anthocyanidin의 효과

이준혁,김귀옥,나형균,박나경,김훈성,김준곤,안지훈,최요한,송시연,배창훈,김용대 대한이비인후과학회 2013 대한이비인후과학회지 두경부외과학 Vol.56 No.5

Background and Objectives Naringenin and delphinidin are types of anthocyanidin, which are flavonoids and thus have anti-inflammatory property. Moderate consumption of natural dietary naringenin and delphinidin is believed to do anti-inflammatory action, but the action mechanism is unclear. Therefore, this study aimed to investigate the effects of naringenin and delphinidin on interleukin-1β (IL-1β)- and lipopolysaccharide (LPS)-induced MUC5AC and MUC5B expressions in airway epithelial cells. Materials and Method In NCI-H292 cells and cultured nasal polyp epithelial cells, the effects of naringenin and delphinidin on IL-1β- and LPS-induced MUC5AC and MUC5B expressions were analyzed by real-time polymerase chain reaction and enzyme-linked immunosorbent assay. Results Delphinidin attenuated IL-1β- and LPS-induced MUC5AC and MUC5B mRNA and glycoprotein expression in a dose-dependent pattern in NCI-H292 cells and in cultured nasal polyp epithelial cells. Naringenin partially attenuated IL-1β- and LPS-induced MUC5AC and MUC5B mRNA and glycoprotein expression at a high dose. Conclusion These results suggest that delphinidin attenuates MUC5AC and MUC5B expressions in the airway epithelial cells. Between anthocyanidin and delphinidin, delphinidin exhibits greater potential as an ideal therapeutic agent for the control of mucus-hypersecretion in the treatment of airway inflammatory diseases.

호흡기 상피세포에서 Roflumilast에 의한 MUC5AC와 MUC5B 발현 억제

김준곤,최윤석,박창휘,이영하,김귀옥,송시연,배창훈,김용대 대한이비인후과학회 2014 대한이비인후과학회지 두경부외과학 Vol.57 No.12

Background and Objectives Roflumilast, a selective inhibitor of phosphodiesterase type 4, has an anti-inflammatory property. It has been used in the treatment of chronic inflammatory airway diseases such as chronic obstructive pulmonary disease and asthma. However, the effect of roflumilast on mucus secretion in inflammatory airway epithelial cells has not been reported. Therefore, this study was aimed at investigating the effects of roflumilast on the inflammatory mediator-induced MUC5AC and MUC5B expression in human airway epithelial cells. Materials and Method In human mucin-producing NCI-H292 airway epithelial cells and primary cultures of nasal epithelial cells, the effects of roflumilast on lipopolysaccharide (LPS)- and phorbl-12-myrsitate-13-acetate (PMA)-induced MUC5AC and MUC5B expression were analyzed by reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay. Results Roflumilast attenuated LPS-induced MUC5AC and MUC5B mRNA and glycoprotein expression in NCI-H292 cells. And roflumilast attenuated PMA-induced MUC5AC and MUC5B mRNA and glycoprotein expression in NCI-H292 cells. In addition, roflumilast attenuated LPS and PMA-induced MUC5AC and MUC5B mRNA expression in the primary cultures of nasal epithelial cells. Conclusion These results suggest that roflumilast attenuates MUC5AC and MUC5B expressions in airway epithelial cells. Roflumilast may be a potentially ideal therapeutic agent for the control of mucus-hypersecretion in treating chronic inflammatory airway diseases. Korean J Otorhinolaryngol-Head Neck Surg 2014;57(12):830-5

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.

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

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

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.

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.

호흡기 상피세포에서 roxithromycin에 의한 MUC5B와 MUC8 점액 유전자 발현 및 점액 생성의 억제

우현재,민명기,배창훈,송시연,김용대 대한이비인후과학회 2008 대한이비인후과학회지 두경부외과학 Vol.51 No.7

Macrolide antibiotics are known to inhibit mucus hypersecretion in patients with chronic airway diseases, but its action mechanism is unclear. Several reports demonstrated that macrolides significantly inhibited gene expression of MUC2, MUC4 and MUC5AC in the airway epithelial cells, but little is known about its inhibitory effect for the other important airway mucins. In upper airway tracts, MUC5B and MUC8 are other important secreted mucin genes. Therefore, this study was aimed to investigate the effects of roxithromycin on the IL-1β-induced gene expression and mucin production of MUC5B and MUC8 in NCI-H292 cells and cultured human nasal polyp epithelial cells. Subjects and Method:The effects of roxithromycin on the IL-1β-induced MUC5B and MUC8 expression were analyzed by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. Results:Roxithromycin attenuated the IL-1β-induced MUC5B and MUC8 gene expression and mucin production with a dose-dependent pattern in NCI-H292 epithelial cells and cultured human nasal polyp epithelial cells. Conclusion:Roxithromycin exerts direct inhibitory effects on the gene expression of MUC5B and MUC8 in airway epithelial cells. These novel findings may explain the clinical efficacy of 14-membered macrolides in the treatment of chronic airway inflammations. (Korean J Otorhinolaryngol-Head Neck Surg 2008;51:617-22)

사람 호흡기 상피세포에서 고농도 포도당 처치 후 활성산소를 통한 MUC5B 점액유전자 발현

이영하,김용대,송유선,배창훈,최윤석,신경철,진현정,송시연 대한이비인후과학회 2017 대한이비인후과학회지 두경부외과학 Vol.60 No.8

Background and Objectives The effects of hyperglycemia on the mucin secretion in inflammatory respiratory diseases are not clear. Therefore, this study was conducted to characterize the effect of hyperglycemia, and the mechanism involved, on MUC5AC and MUC5B expression in human airway epithelial cells. Materials and Method The NCI-H292 cells and the primary cultures of human nasal epithelial cells were exposed to different concentration of glucose (5, 10, 15, 20, 30 mM) for 8 or 24 hours, the effects of high concentration of glucose (20 mM) on MUC5AC and MUC5B expression were determined using reverse transcriptase-polymerase chain reaction (PCR), real-time PCR and enzyme immunoassay. Measurement of reactive oxygen species (ROS) production was performed by flow cytometry. To investigate the role of ROS in high concentration of glucose-induced MUC5B expression, the cells were pretreated with N-acetyl-cysteine (NAC, 50 mM) as a ROS scavenger, or diphenyleneiodonium (DPI, 100 nM) as a nicotinamide adenine dinucleotide phosphate oxidase inhibitor for 1 hour. Results In the NCI-H292 cells and the primary cultures of human nasal epithelial cells, High concentration of glucose increased MUC5B expression but did not increase MUC5AC expression (p<0.05). ROS production was also increased by high concentration of glucose (20 mM) (p<0.05). In addition, high concentration of glucose (20 mM)-induced MUC5B expression and ROS production were significantly attenuated by pretreatment of NAC (50 mM) or DPI (100 nM) (p<0.05). Conclusion High concentration of glucose induces MUC5B expressions via ROS in human airway epithelial cells. Korean J Otorhinolaryngol-Head Neck Surg 2017;60(8):396-403