Dickkopf-1 inhibits the secretion of MUC5AC induced by Mycoplasma pneumoniae P1-C in mouse lung epithelial cells / 生物工程学报

Mycoplasma pneumoniae is the most common pathogen of respiratory tract infection in children and adults. Clinical observation shows that M. pneumoniae infection can cause massive mucus secretion in the respiratory tract, which makes the breathing of patients difficult. Studies have shown that M. pneumoniae infection can cause massive secretion of mucin 5AC (MUC5AC). Adhesin P1 plays an important role in the pathogenesis of M. pneumoniae infection by mediating the adhesion of pathogens to host cells, and the C-terminal residues of P1 (P1-C) are immunogenic. This study investigated the molecular mechanism of Wnt/β-catenin signaling pathway inhibitor Dickkopf-1 (DKK1) in the secretion of MUC5AC in mouse airway epithelial cells (MAECs) induced by P1-C. Scanning electron microscope and hematoxylin-eosin staining were used to observe the effect of P1-C on mucus secretion of MAECs. Protein chip was used to detect the secretion of cytokines and analyse the enrichment of related signaling pathways induced by P1-C in MAECs. Periodic acid schiff stain (PAS) staining, Tunel staining and Masson staining were used to detect the damage of the lungs of mouse exposed to P1-C. Immunohistochemistry was used to detect the secretion of MUC5AC expression, and Western blotting was used to reveal the molecular mechanism of DKK1-regulated secretion of MUC5AC induced by P1-C protein in MACES. The results showed that P1-C induced the massive secretion of mucus and inflammatory factors in MAECs. During P1-C infection, DKK1 down-regulated janus kinase 2 (JAK2), phosphorylation signaling and transcription activator 1 (p-STAT1) and phosphorylation signaling and activator of transcription 3 (p-STAT3) expression. Overexpression of DKK1 significantly up-regulated the expression of MUC5AC repressor transcription factor fork-head box protein A2 (FOXA2). At the same time, the expression of MUC5AC induced by P1-C was inhibited significantly. It is speculated that DKK1 can effectively reduce the secretion of MUC5AC in MAECs induced by P1-C by inhibiting the JAK/STAT1-STAT3 signaling pathway and up-regulating the expression of FOXA2.

Increased expression of pendrin in eosinophilic chronic rhinosinusitis with nasal polyps / Expressão aumentada de pendrina na rinossinusite crônica eosinofílica com pólipos

Abstract Introduction: Chronic rhinosinusitis with nasal polyps is a heterogeneous disease and appropriate diagnostic algorithms in individual cases are necessary for effective medical treatment. Objective: The purpose of this study was to clarify the relationship between the pendrin expression of nasal polyps and clinical and pathological characteristic features of eosinophilic chronic rhinosinusitis. Methods: A total of 68 patients were classified into eosinophilic chronic rhinosinusitis or non-eosinophilic chronic rhinosinusitis groups according to the degree of eosinophilic infiltration into the nasal polyps. Clinical, hematological, and immunohistochemical analyses were performed and statistically compared between both groups. Results: Thirty-eight were classified into eosinophilic chronic rhinosinusitis and 30 into non-eosinophilic chronic rhinosinusitis groups. There were no significant differences in age distribution, sex ratio, prevalence of asthma, or any other complications between the groups. The mean Lund-Mackay score and the number of serum eosinophils was significantly higher in the eosinophilic chronic rhinosinusitis than in the non-eosinophilic chronic rhinosinusitis groups. The pendrin expression was more frequently detected in the epithelial surface layer of nasal polyps in the eosinophilic chronic rhinosinusitis than in the non-eosinophilic chronic rhinosinusitis groups. In addition, mucin 5AC was more widely expressed in the eosinophilic chronic rhinosinusitis than in the non-eosinophilic chronic rhinosinusitis. Conclusion: Increased expression of pendrin and mucin 5AC in the nasal polyps would be associated with development of eosinophilic chronic rhinosinusitis. This finding could allow the development of a novel therapeutic agent targeted specifically to patients with eosinophilic chronic rhinosinusitis.

Resumo Introdução: A rinossinusite crônica com pólipos nasais é uma doença heterogênea e algoritmos diagnósticos apropriados em casos individuais são necessários para um tratamento médico eficaz. Objetivo: O objetivo deste estudo foi esclarecer a relação entre a expressão da pendrina de pólipos nasais e propriedades clínicas e patológicas características da rinossinusite crônica eosinofílica. Método: Um total de 68 pacientes foram classificados como tendo rinossinusite crônica eosinofílica ou rinossinusite crônica não eosinofílica de acordo com o grau de infiltração eosinofílica nos pólipos nasais. Análises clínicas, hematológicas e imunohistoquímicas foram realizadas e comparadas estatisticamente entre os dois grupos. Resultados: Entre os pacientes, 38 apresentavam rinossinusite crônica eosinofílica e constituíram o grupo 1; 30 tinham rinossinusite crônica não eosinofílica e constituíram o grupo 2. Não houve diferenças significantes na distribuição etária, razão entre os sexos, prevalência de asma ou qualquer outra complicação entre os grupos. O escore médio de Lund-Mackay e o número de eosinófilos séricos foram significantemente maiores no grupo com rinossinusite crônica eosinofílica do que no grupo com rinossinusite crônica não eosinofílica. A expressão da pendrina foi mais frequentemente detectada na camada epitelial superficial dos pólipos nasais na rinossinusite crônica eosinofílica do que no grupo com rinossinusite crônica não eosinofílica. Além disso, mucina 5AC foi mais amplamente expressa na rinossinusite crônica eosinofílica do que na rinossinusite crônica não eosinofílica. Conclusão: O aumento da expressão da pendrina e mucina 5AC nos pólipos nasais estaria associado ao desenvolvimento de rinossinusite crônica eosinofílica. Esse achado pode permitir o desenvolvimento de um novo agente terapêutico voltado especificamente para pacientes com rinossinusite crônica eosinofílica.

Expression of MUC5AC in normal gastric mucosa, intestinal metaplasia and gastric carcinoma by immunohistochemistry

Background: Carcinomas of the stomach are a heterogeneous group of lesions in terms of architecture, pattern of growth, cell differentiation, and histogenesis. Altered MUC5AC expression patterns have been reported previously in intestinal metaplasia as well as in gastric cancer. The aim of the study was to analyse the expression pattern of MUC5AC in normal, pre-neoplastic and neoplastic gastric epithelium.Methods: Formalin fixed paraffin embedded sections of sixty cases which include twenty cases of each normal gastric mucosa, intestinal metaplasia and gastric carcinoma were taken up for the study and subjected to immunohistochemistry using MUC5AC.Results: The intensity of MUC5AC immunostaining in normal gastric mucosa, intestinal metaplasia and gastric carcinoma was evaluated. Immunoreactivity was graded as 0 (negative), ± (trace positive), + (positive) or ++ (strongly positive). Statistical analysis was performed with Chi-Square test and significant differences were noted between these 3 groups (p value <0.05).Conclusions: Authors concluded that MUC5AC expression rates might be good parameters in progression of intestinal metaplasia to gastric carcinoma and might be a good prognostic marker for gastric carcinoma as it is very well implicated in understanding of gastric carcinogenesis.

Effect of Resolvin D1 and E1 on Mucin Expression in Human Airway Epithelial Cells / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Mucin is an important component of mucus that performs the first line of defense against inhaled pathogens and particles, lubrication of organs, and protection of airway. It is hyper-secreted in inflammatory airway diseases and is associated with morbidity and mortality of the affected patients. Resolvin, an autacoid of a specific lipid structure, exhibits anti-inflammatory property against inflammatory airway diseases although its effects 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)-induced mucin expression in human airway epithelial cells. MATERIALS AND METHOD: In mucin-producing human NCI-H292 epithelial cells, the effects and brief signaling pathways of Resolvin D1 (RvD1) and Resolvin E1 (RvE1) on the LPS-induced MUC4, 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 expression and protein production in human NCI-H292 cells while RvE1 did not. RvD1 significantly blocked 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 in airway epithelial cells. Therefore, RvD1 may modulate the control of mucus-hypersecretion in inflammatory airway diseases.

Effect of Pinelliae Rhizoma Praeparatum Cum Alumine Polysaccharides on MUC5AC mRNA in Lung Tissues of Allergic Asthma Model Rats / 中国实验方剂学杂志

Objective: To observe the effect of Pinelliae Rhizoma Praeparatum Cum Alumine polysaccharides(PRPCAP)on airway mucus secretion in rats with allergic asthma, in order to study the material basis of the "macromolecule" component of the polysaccharides as the original medicinal materials. Method: The 60 SPF-grade Wistar rats were induced by intraperitoneal injection of chicken ovalbumin (OVA) and aluminum-magnesium adjuvant, except for the normal control group. The OVA solution was aerosolized to establish a rat model of allergic asthma. After successful modeling, the rats were randomly divided into 5 groups, namely allergic asthma model group, positive drug group (montalurast sodium,5 mg·kg-1), high-dose PRPCAP group (400 mg·kg-1), middle-dose PRPCAP group (200 mg·kg-1) and low-dose PRPCAP group (100 mg·kg-1). The contents of interleukin-4 (IL-4) and interferon-γ (IFN-γ) in serum and bronchoalveolar lavage fluid (BALF) supernatant were determined by enzyme-linked immunosorbent assay (ELISA), and the count of eosinophils (EOS) was detected by BALF sediment. The histopathological changes were observed by hematoxylin-eosin (HE) staining in lung tissue. The mRNA expression of mucin 5AC (MUC5AC) was detected by Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR). Result: Compared with the normal control group, serum IL-4 level in the allergic asthma model group was significantly increased (Pγ level was significantly decreased (PPPPPγ in serum (PPPPConclusion: The "macromolecule" component of polysaccharides in the Pinelliae Rhizoma Praeparatum Cum Alumine may be the material basis for the efficacy of eliminating dampness and eliminating phlegm.

Changes in EGFR, FOXA2 and MUC5AC expression in respiratory syncytial virus-infected human bronchial epithelium cells and effects of rosiglitazone intervention / 中华微生物学和免疫学杂志

Objective To investigate the mechanism of epidermal growth factor receptor-forkhead transcription factor A2 (EGFR-FOXA2) pathway-involved high secretion of mucus in human bronchial epitheli-um (HBE) cells after respiratory syncytial virus (RSV) infection and to evaluate the effects of intervention using agonist ( rosiglitazone ) and antagonist ( GW9662 ) of peroxidase proliferation activated receptor γ( PPARγ) and EGFR inhibitor ( AG1478 ) . Methods HBE cells were randomly divided into six groups: A group ( AG1478+RSV) , B group ( rosiglitazone+RSV) , C group ( GW9662+RSV) , D group ( RSV) , E group (0. 1％ dimethyl sulfoxide DMSO) and F group (HBE cell control group). Two hours before RSV infection, A, B and C groups were respectively treated with 10 μmol/L of AG1478, rosiglitazone and GW9662. Expression of EGFR, PPARγ and FOXA2 at mRNA level in each group was detected by real-time fluorescence quantitative RT-PCR 12 h, 24 h and 48 h after HBE cells were infected with or without RSV. Expression of phosphorylated-EGFR ( p-EGFR) and EGFR at protein level was detected by Western blot. ELISA was performed to measure the expression of mucin-5AC (MUC5AC). Results Compared with F group, EGFR expression at mRNA lev-el, p-EGFR/EGFR protein ratio and MUC5AC expression at protein level were increased in a time-dependent manner in A, B, C and D groups at 12 h, 24 h and 48 h. Compared with group F, the expression of PPARγat mRNA level in A, B, and D groups increased at each time point. Moreover, PPARγ expression gradually in-creased over time in A and B groups, reaching the peaks at 48 h, but was in decline in D group. Expression of FOXA2 at mRNA level in RSV-infected HBE cells was declined at each time point compared with that in group F, especially in D group. Compared with group D, A and B groups showed significantly decreased EGFR ex-pression at mRNA level, p-EGFR/EGFR protein ratio and MUC5AC expression at protein level, but markedly increased FOXA2 expression at mRNA level. Conclusions RSV infection increased the expression of MUC5AC at protein level in HBE cells. PPARγand EGFR-FOXA2 signaling pathways were involved in the hypersecretion of airway mucus during RSV infection.

Inhibitory Effects of Protopanaxadiol on Lipopolysaccharide-Induced Reactive Oxygen Species Production and MUC5AC Expression in Human Airway Epithelial Cells / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: MUC5AC is one of the major secretory mucin genes in the human 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 have anti-inflammatory, antitumor and antioxidant properties. However, the effects of PPD on mucin secretion of airway epithelial cells still have not been reported. Therefore, the aim of this study is to investigate the effect of PPD on lipopolysaccharide (LPS)-induced MUC5AC expression in 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-polymerase chain reaction and enzyme immunoassay after treated with LPS. N-acetylcysteine (NAC) as a reactive oxygen species (ROS) scavenger, and apocynin as a nicotinamide adenine dinucleotide phosphate oxidase inhibitor were used to compare the inhibitory effect of PPD on LPS-induced ROS 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 expression and protein production as well as ROS production. In addition, NAC and apocynin inhibited LPS-induced MUC5AC mRNA expression and protein production. CONCLUSION: These results demonstrate that PPD inhibits LPS-induced MUC5AC expression via ROS in human airway epithelial cells and the inhibitory effect of PPD was similar to that of NAC and apocynin. These findings indicate that PPD may be a therapeutic agent for control of mucus secretion and oxidative stress in human airway epithelial cells.

Role of inositol-requiring kinase 1α/X-box binding protein 1 in airway mucus secretion induced by neutrophil elastase / 上海交通大学学报（医学版）

Objective: To explore the effect of endoplasmic reticulum stress (ERS) on neutrophil elastase (NE) induced mucin 5AC (MUC5AC) production in human airway epithelial cells. Methods: HBE16 airway epithelial cells were cultured and pretreated with reactive oxygen species (ROS) inhibitor N-acetylcysteine (NAC) or ERS inhibitor 4-phenylbutyrate (4-PBA), or transfected with small interfering RNA (siRNA) against inositolrequiring kinase 1α (IRE-1α) or X-box binding protein 1 (XBP-1), respectively before incubation with NE. NE group and blank control group were also set up. ROS production was assayed by detection kit; expression of glucose-regulated protein 78 (GRP78), phosphorylated protein kinase R-like endoplasmic reticulum kinase (pPERK), activating transcription factor 6 (ATF6), phosphorylated IRE-1α (pIRE-1α), and XBP-1 protein was detected by Western blotting; spliced XBP-1 (XBP-1s) mRNA was measured by real-time PCR; levels of MUC5AC protein in culture supernatant and cytoplasm were assayed by ELISA and immunofluorescence. Results: There was an obvious increase of ROS production with strong elevation of GRP78, ATF6, pPERK, and pIRE-1α protein in NE group cells after 24 h, compared with blank control group (P<0.05). The protein and mRNA of XBP-1s, and MUC5AC production also increased obviously (P<0.05). NAC and 4-PBA reduced ERS-related protein expression and MUC5AC production and secretion (P<0.05). Further studies showed that MUC5AC secretion was also blunted by IRE-1α siRNA or XBP-1 siRNA, accompanied with decreased expression of XBP-1s mRNA and protein (P<0.05). Conclusion: NE induces ERS by producing ROS, and increases MUC5AC protein production and secretion; IRE-1α/XBP-1 play a certain role in this process.

Triptolide Inhibits Lipopolysaccharide-Induced MUC5AC/5B Expression via Nuclear Factor-Kappa B in Human Airway Epithelial Cells / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: The representative mucin genes in the human airway are MUC5AC and MUC5B, which are regulated by several inflammatory and anti-inflammatory substances. Triptolide (TPL), udenafil, betulinic acid, changkil saponin, and glucosteroid are some of the many anti-inflammatory substances that exist. TPL is a diterpenoid compound from the thunder god vine, which is used in traditional Chinese medicine for treatment of immune inflammatory diseases, such as rheumatoid arthritis, systemic lupus erythematosus, nephritis and asthma. However, the effects of TPL on mucin expression of human airway epithelial cells have 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 epithelial cells were used to investigate the effects of TPL on LPS-induced MUC5AC and MUC5B expression using real-time polymerase chain reaction, enzyme immunoassay, and Western blot. RESULTS: TPL significantly decreased the LPS-induced MUC5AC and MUC5B mRNA expression and 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 expression via inhibition of NF-kB activation in human airway epithelial cells. This study may provide important information about the biological role of triptolide on mucus-secretion in airway inflammatory diseases and the development of novel therapeutic agents for controlling such diseases.

Effect of Lyn on expression of MUC5AC in human bronchial epithelial cells induced by house dust mite and its mechanism / 吉林大学学报(医学版)

Objective:To investigate the effect of Lyn on the expression of MUC5AC in human bronchial epithelial cells induced by house dust mite(HDM),and to explore its possible mechanism.Methods:The human bronchial epithelial cells(16 HBE)were divided into PBS group and HDM group(1 μg·L-1HDM).The cells were transfected by liposome.The luciferase report gene of MUC5AC promoter was constructed.The relative luciferase unit(RLU)was detected by double luciferase report gene assay.The expression of MUC5AC in the cells was detected by immunofluorescence technique and observed by confocal microscope.The expression level of STAT6 in the 16HBE cells was detected by Western blotting method.Results:The results of double luciferase report gene assay showed that the RLU in HDM group was higher than that in PBS group(P<0.05).The RLU of cells in HDM group treated with LynsiRNA intervention was higher than that of the cells without LynsiRNA intervention(P<0.05).The immunofluorescence results demonstrated that the expression level of MUC5AC in HDM group was higher than that in PBS group(P<0.05),and the expression level of MUC5AC in HDM group was increased after LynsiRNA intervention(P<0.05).The Western blotting results indicated that the expression level of STAT6 was up-regulated in HDM group when the cells were intervened with LynsiRNA(P<0.05). Conclusion:Deficiencyof Lyn can increase the expression of MUC5AC in the human bronchial epithelial cells,and its mechanism may be related to regulating the STAT6 signal pathway by Lyn.

Effect of Lyn on expression of MUC5AC in human bronchial epithelial cells induced by house dust mite and its mechanism / 吉林大学学报(医学版)

Objective: To investigate the effect of Lyn on the expression of MUC5AC in human bronchial epithelial cells induced by house dust mite (HDM), and to explore its possible mechanism. Methods: The human bronchial epithelial cells 16HBE) were divided into PBS group and HDM group 1 μg · L-1 HDM). The cells were transfected by liposome. The luciferase report gene of MUC5AC promoter was constructed. The relative luciferase unit (RLU) was detected by double luciferase report gene assay. The expression of MUC5AC in the cells was detected by immunofluorescence technique and observed by confocal microscope. The expression level of STAT6 in the 16HBE cells was detected by Western blotting method. Results: The results of double luciferase report gene assay showed that the RLU in HDM group was higher than that in PBS group (P<0.05). The RLU of cells in HDM group treated with LynsiRNA intervention was higher than that of the cells without LynsiRNA intervention (P<0.05). The immunofluorescence results demonstrated that the expression level of MUC5AC in HDM group was higher than that in PBS group (P<0.05), and the expression level of MUC5AC in HDM group was increased after LynsiRNA intervention (P<0.05). The Western blotting results indicated that the expression level of STAT6 was up-regulated in HDM group when the cells were intervened with LynsiRNA (P<0.05). Conclusion: Deficiency of Lyn can increase the expression of MUC5AC in the human bronchial epithelial cells, and its mechanism may be related to regulating the STAT6 signal pathway by Lyn.

Clusterin Induces MUC5AC Expression via Activation of NF-κB in Human Airway Epithelial Cells

OBJECTIVES: Clusterin (CLU) is known as apolipoprotein J, and has three isoforms with different biological functions. CLU is associated with various diseases such as Alzheimer disease, atherosclerosis, and some malignancies. Recent studies report an association of CLU with inflammation and immune response in inflammatory airway diseases. However, the effect of CLU on mucin secretion of airway epithelial cells has not yet been understood. Therefore, the effect and brief signaling pathway of CLU on MUC5AC (as a major secreted mucin) expression were investigated in human airway epithelial cells. METHODS: In the tissues of nasal polyp and normal inferior turbinate, the presence of MUC5AC and CLU was investigated using immunohistochemical stain and Western blot analysis. In mucin-producing human NCI-H292 airway epithelial cells and primary cultures of normal nasal epithelial cells, the effect and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway of CLU on MUC5AC expression were investigated using immunohistochemical stain, reverse transcription-polymerase chain reaction, real-time polymerase chain reaction, enzyme immunoassay, and Western blot analysis. RESULTS: In the nasal polyps, MUC5AC and CLU were abundantly present in the epithelium on immunohistochemical stain, and nuclear CLU (nCLU) was strongly detected on Western blot analysis. In human NCI-H292 airway epithelial cells or the primary cultures of normal nasal epithelial cells, recombinant nCLU increased MUC5AC expression, and significantly activated phosphorylation of NF-κB. And BAY 11-7085 (a specific NF-κB inhibitor) and knockdown of NF-κB by NF-κB siRNA (small interfering RNA) significantly attenuated recombinant nCLU-induced MUC5AC expression. CONCLUSION: These results suggest that nCLU induces MUC5AC expression via the activation of NF-κB signaling pathway in human airway epithelial cells.

SOCS3 regulates mucus hypersecretion via JAK/STAT signal pathway / 安徽医科大学学报

Objective To investigate the SOCS3 regulates mucus hypersecretion via JAK/STAT signal pathway in inflammatory cells.Methods Culture 16HBE cells and divide into three groups:control group,inter leukin(IL)-6-exposed group;IL-6-exposed and microRNA-203-transfered group.The protein levels of JAK1/2,SOCS3 and MUC5AC were measured by Western blot.The mRNA expressions of SOCS3,STAT3 and MUC5AC were detected by Real-time PCR.The protein levels of p-JAK1/2,SOCS3 and MUC5AC were analyzed by ELISA.Results Compared with the control group,the mRNA expressions of SOCS3,STAT3,MUC5AC and the protein levels of p-JAK1/2,SOCS3,MUC5AC were both significantly increased in IL-6-exposed group (P ＜ 0.05);in the IL-6-exposed and miR-203-transfered group,the protein levels of p-JAK1/2,MUC5AC and the mRNA expressions of STAT3 were both significantly increased (P ＜ 0.05),but the mRNA expressions of SOCS3 was almost as much as that in IL-6-exposed group,the protein levels of SOCS3 was significantly decreased (P ＜ 0.05).Conclusion SOCS3 over-express when cells are stimulated by IL-6,and negative-feedback regulates MUC5AC secretion via JAK/STAT signal pathway.

Expression levels of HIF-1 α and mRNA and MUC5AC,MUC5B mRNA in chronic rhinosinusitis and their correlation / 国际检验医学杂志

Objective To explore the expression levels of HIF-1 α and MUC5AC,MUC5B mRNA in chronic rhinosinusitis(CRS) and their correlation.Methods Eighty cases of CRS with nasal polyps(CRSwNP) treated in our hospital from May 2014 to May 2015,80 cases of CRS without NP(CRSsNP) and 80 healthy people without CRS(control group) were chosen.Nasal sinus mucosa in 3 groups were collected and treated.The expressions of HIF-1 α,MUC5AC and MUC5B mRNA were measured by semiquantitative reverse transcription polymerase chain reaction(RT-PCR),and their correlations were analyzed.Results The relative expression levels of HIF-1 α,MUC5AC and MUC5B mRNA in the CRSwNP group or CRSsNP group were 1.35±0.85,1.35±0.91;0.80±0.55,0.79±0.49 and 1.18±1.01,1.21±1.02 respectively,while which in the control group were 0.42±0.33,0.43±0.36 and 0.47±0.43 respectively,the difference between the CRS groups and control group was statistically significant(P<0.05).The expression of HIF-1αin the CRSwNP group and CRSsNP group was positively correlated with MUC5AC,MUC 5B mRNA(r=0.476,P=0.023,r=0.476,P=0.026,r=0.478,P=0.035,r=0.508,P=0.021).The ROC analysis results showed AUC=0.956 4.Conclusion The expression levels of HIF-1α and MUC5AC,MUC5B mRNA in chronic rhinosinusitis are significantly upregulated,moreover HIF-1α is closely related with MUC5AC and MUC5B mRNA.

Effects of Homogentisic Acid and Natural Products Derived from Pinellia ternata on Secretion, Production and Gene Expression of MUC5AC Mucin from Cultured Airway Epithelial Cells

In this study, we investigated whether adenosine, adenine, uridine and homogentisic acid derived from Pinellia ternata affect the secretion, production and gene expression of MUC5AC mucin from airway epithelial cells. Confluent NCI-H292 cells were pretreated with adenosine, adenine, uridine or homogentisic acid for 30 min and then stimulated with PMA (phorbol 12-myristate 13-acetate) for 24 h. The MUC5AC mucin gene expression, mucin protein production and secretion were measured by RT-PCR and ELISA, respectively. The results were as follows: (1) Adenine and homogentisic acid decreased PMA-induced MUC5AC mucin gene expression, although adenosine and uridine did not affect the mucin gene expression; (2) Adenosine, adenine, uridine and homogentisic acid inhibited PMA-induced MUC5AC mucin production; (3) Homogentisic acid inhibited the secretion of MUC5AC mucin from NCI-H292 cells. These results suggest that, among the four compounds examined, homogentisic acid showed the regulatory effect on the steps of gene expression, production and secretion of mucin, by directly acting on airway epithelial cells.

Mometasone Furoate Suppresses PMA-Induced MUC-5AC and MUC-2 Production in Human Airway Epithelial Cells / 결핵및호흡기질환

BACKGROUND: Mucus hypersecretion from airway epithelium is a characteristic feature of airway inflammatory diseases. Tumor necrosis factor α (TNF-α) regulates mucin synthesis. Glucocorticoids including mometasone fuorate (MF) have been used to attenuate airway inflammation. However, effects of MF on mucin production have not been reported. METHODS: Effects of MF and budesonide (BUD) on the phorbol-12-myristate-13-acetate (PMA)–induction of mucin and TNF-α in human airway epithelial cells (NCI-H292) were investigated in the present study. Confluent NCI-H292 cells were pretreated with PMA (200 nM) for 2 hours. Subsequently, the cells were stimulated with MF (1–500 ng/mL) or BUD (21.5 ng/mL) for 8 hours. Dexamethasone (1 µg/mL) was used as the positive control. Real-time polymerase chain reaction was used to determine MUC2 and MUC5AC mRNA levels. The level of total mucin, MUC2, MUC5AC, and TNF-α in culture supernatants were measured using enzyme-linked immunosorbent assay. RESULTS: MF and BUD significantly suppressed MUC2 and MUC5AC gene expression in PMA-stimulated NCI-H292 cells. The inhibitory effects of the two steroid drugs were also observed in the production of total mucin, MUC2 and MUC5AC proteins, and TNF-α. CONCLUSION: Our findings demonstrated that MF and BUD attenuated mucin and TNF-α production in PMA-induced human airway epithelial cells.

Effects of RASI on expression of airway MUC5 AC and related inflammatory factors in asthmatic mice with Yin deficiency syndrome / 中国免疫学杂志

Objective:To study the effects of Radix Angelicae Sinensis ( RASI) on expression of airway MUC5AC and related inflammatory factors in asthmatic mice with Yin deficiency syndrome.Methods:Injecting ovalbumin ( OVA) to sensitize,inhaling OVA to stimulate,using Thyroxin during late stimulation,the asthmatic mouse with Yin deficiency syndrome was established and evaluated through asthmatic behaviors, lung histopathology, active factors ( IL-13, TNF-αand MUC5AC ) in broncho-alveolar lavage fluid (BALF), and MUC5AC expression in lung tissue.Results: 2,4,8 g/kg RASI could reduce asthmatic behaviors score, relieve pathological changes of lung tissue,reduce the contents of IL-13,TNF-αand MUC5AC in BALF,and depress MUC5AC expression in lung tissue ( P<0.05,0.01).In addition,there was a certain synergy between RASI and dexamethasone ( DXM) on depressing the ex-pression of IL-13 and MUC5AC (P<0.05).Conclusion:RASI has certain anti-asthma effect and one of mechanisms is to regulate the MUC5AC expression through inhibit IL-13 and TNF-α.On the expression of IL-13 and MUC5AC,the compatibility of RASI with glu-cocorticoid has some synergy effect.

Expression of the VEGF and mucin MUC5AC in nasal mucosa before and after chronic rhinitis - sinusitis and nasal polyps - endoscopic sinus surgery / 中国内镜杂志

Objective To investigate the expression of the vascular endothelial growth factor (VEGF) and mucin MUC5AC in nasal mucosa before and after chronic rhinitis - sinusitis and nasal polyps - endoscopic sinus surgery.Methods 75 cases chronic rhinitis - sinusitis and nasal polyps - endoscopic sinus surgery patients were selected as nasal polyps group and 75 cases of nasal bone fracture or epistaxis patients as the control group from January 2012 to January 2015. Took the samples of nasal polyps before surgery and the maxillary sinus mucosa specimens after surgery six weeks of nasal polyps’ patients and on the edge of the inferior turbinate mucosa specimens of the control group to detect eosinophil count by HE staining, and detect the expression of VEGF and mucin MUC5AC by immunohistochemical staining.Results The specimens eosinophils of preoperative nasal polyp group and postoperative nasal polyp group were higher than that of control group (P < 0.05), the nasal eosinophils of postoperative nasal polyp group was lower than that of preoperative nasal polyp group (P < 0.05). The expression of specimens VEGF and mucin MUC5AC area percentages in preoperative nasal polyp group and postoperative nasal polyp group were higher than that in control group (P < 0.05), the expression of nasal VEGF and mucin MUC5AC area percentages in the postoperative nasal polyp group was lower than that of preoperative nasal polyp group (P< 0.05).Conclusion Eosinophil count and the expression levels of VEGF and mucin MUC5AC of nasal mucosa in chronic rhinitis - sinusitis and nasal polyps - endoscopic sinus surgery preoperative are higher, and reduce at postoperative six weeks, VEGF and mucin MUC5AC may be involved nasal repair.

Effects ofJiedu QingfeiMixture on Expressions of NE and MUC5AC in Lung Tissue of Rats with Chronic Obstructive Pulmonary Disease / 中国中医药信息杂志

Objective To explore the mechanism ofJiedu Qingfei Mixture for airway mucus hypersecretion of rat models with chronic obstructive pulmonary disease (COPD).Methods Airway instilling lipopolysaccharide combining fuming method was used to establish COPD models. Forty clean level Wistar strain rats were randomly divided into blank control group, model group,Jiedu Qingfei group, and clarithromycin group. Model group, Jiedu Qingfei group, and clarithromycin group were given normal saline,Jiedu Qingfei Mixture, and clarithromycin by gavage respectively, while the blank control group was raised normally for 30 d. All rats were killed on the 31st day for taking lung tissue (6 rats from each group were chosen randomly). Pathological changes of lung tissue and mucous glands hyperplasia were observed by HE staining method. NE and MUC5AC mRNA expression on lung tissue were detected by RT-PCR method. Protein expressions of NE and MUC5AC on pulmonary tissue and airway epithelium were detected by immunohistochemical method.Results Compared with blank control group, mucous glands hyperplasia on airway epithelium, mRNA expression of NE and MUC5AC in lung tissue, and protein expressions of NE and MUCA5C on airway epithelium in the model group significantly increased (P<0.05,P<0.01). Compared with model group, mucous glands hyperplasia on airway epithelium inJiedu Qingfei group significantly decreased (P<0.01), as same as clarithromycin group;Jiedu Qingfei group showed better effects on down-regulating NE and MUC5AC mRNA expression in lung tissue compared with clarithromycin group. MUC5AC protein expression on airway epithelium inJiedu Qingfei group significantly decreased (P<0.05), as same as clarithromycin group.Jiedu Qingfei group and clarithromycin group had no difference on NE protein expression in airway epithelium compared with model group.Conclusion Jiedu Qingfei group Mixture can reduce airway mucus hypersecretion of COPD by down-regulating MUC5AC expression through neutrophil elastase.

Effects of bencycloquidium bromide on the expression of MUC5 AC induced by lipopolysaccharide in cultured human nasal epithelial cells / 中国药理学通报

Aim Toinvestigatetheeffectofbencyclo-quidium bromide(BCQB)on mucus MUC5AC expres-sion induced by lipopolysaccharide in cultured human nasalepithelialcells(HNECs).Methods Primary culture of human nasal epithelial cells (HNECs)was randomly divided into control group (C,with no treat-ment),LPS group (LPS,with LPS 1 mg · L-1 added in),BCQB low dose group(BCQBL,with LPS 1 mg· L-1 and BCQB 10 -8 mol·L-1 added in),BCQB mid-dle dose group(BCQBM,with LPS 1 mg·L-1 and BC-QB 10 -7 mol·L-1 added in),BCQB high dose group (BCQBH,with LPS 1 mg·L-1 and BCQB 10 -6 mol· L-1 added in)and ipratropium bromide group(IB,with LPS 1 mg·L-1 and IB 10 -6 mol·L-1 added in).Af-ter incubation at 37 ℃with 5% CO2 for 24 h,the ex-pression of MUC5 AC mRNA was detected with Real-time PCR and the expression of MUC5 AC protein in HNECs was detected with Western blot,while the ex-pression of MUC5 AC protein in supernatant was detec-tedwithELISAineachgroup.Results Ascompared with control group,the expression of MUC5 AC mRNA and protein increased significantly in LPS group (each P0. 05 ). Conclusion Bencycloquidiumbromidecansuppress MUC5 AC expression induced by LPS in cultured hu-man nasal epithelial cells,indicating that BCQB may be a new drug for nasal mucous hypersecretion diseases.