Objective:To explore the role of NLRP3 in mucus hypersecretion in asthmatic patients.Methods:From January 2020 to June 2022,90 patients with asthma and 60 healthy patients under the Department of Pulmonary and Critica...Objective:To explore the role of NLRP3 in mucus hypersecretion in asthmatic patients.Methods:From January 2020 to June 2022,90 patients with asthma and 60 healthy patients under the Department of Pulmonary and Critical Care Medicine of the First Affiliated Hospital of Xi’an Medical University were selected.Immunohistochemistry and enzyme-linked immunosorbent assay were performed.NLRP3 inflammasome and mucins MUC5AC and MUC5B levels in lung tissue and sputum were detected.Results:Compared to the healthy control group,the asthma group had significantly higher sputum MUC5A(20.12±5.07 versus 36.21±6.13)and NLRP3(72.31±15.13 versus 119.21±31.21)levels(P<0.05)but lower MUC5B levels(1.35±0.12 versus 0.53±0.11,P<0.05).Immunohistochemistry showed that NLRP3,MUC5AC,and MUC5B expressions were consistent with the sputum results.Conclusion:NLRP3 and MUC5AC levels are significantly increased in asthmatic patients,whereas MUC5B levels are reduced in these patients.They can be used as targets for the diagnosis and treatment of asthma.展开更多
In the human airway, the gelforming mucin subtypes MUC5B and MUC5AC play important roles in biophylaxis. However, the regulation of MUC5B production is less clear than that of MUC5AC. Therefore, the regulation of MUC5...In the human airway, the gelforming mucin subtypes MUC5B and MUC5AC play important roles in biophylaxis. However, the regulation of MUC5B production is less clear than that of MUC5AC. Therefore, the regulation of MUC5B production by cell attachment and Akt was investigated in human lung epithelial NCI-H292 cells. We found that low cell attachment to culture plates induced the upregulated production of both MUC5B and MUC5AC. Cell attachment induces the activation of Akt, a serine/threonine kinase. Cell treatment with Akt inhibitor I decreased Akt phosphorylation and activation. However, MUC5B production was unaffected by Akt inhibition, whereas MUC5AC production was upregulated. MUC5B production was also unaffected by Akt inhibition in cells cultured on type IV collagen or fibronectin. These results suggest that the production of both MUC5B and MUC5AC is regulated by cell attachment. However, the regulation of MUC5B is unaffected by Akt inhibition, in contrast to that of MUC5AC.展开更多
Glycosylation of mucins mediated by N-acetylgalactosaminyltransferases(GALNTs)is closely related to respiratory diseases such as asthma and chronic obstructive pulmonary disease(COPD).In addition,long non-coding RNAs(...Glycosylation of mucins mediated by N-acetylgalactosaminyltransferases(GALNTs)is closely related to respiratory diseases such as asthma and chronic obstructive pulmonary disease(COPD).In addition,long non-coding RNAs(LncRNAs)participate in physiological and pathological processes through various epigenetic mechanisms.In this study,we found that a novel LncRNA named NKILA combined with multiple mucins and GALNTs potentially by several bioinformatics methods,and we used quantitative real-time PCR(RT-qPCR)to detect the expressions of NKILA,MUC5AC,MUC5B,and GALNT2 mRNA in 50 cases of asthma samples and 19 cases of normal samples,whose results showed that the expression of NKILA was significantly decreased in asthmatic samples,negatively correlated with the severity of asthma and the expressions of MUC5AC and MUC5B,while GALNT2 was significantly increased in asthmatic tissues,and positively correlated with the severity of asthma and the expressions of MUC5AC and MUC5B.In vitro,we used transient transfection technology to overexpress or interfere with NKILA and GALNT2 and then detected the expressions of MUC5AC and MUC5B via RT-qPCR and Western blot,which demonstrated GALNT2 can promote the expressions of MUC5AC and MUC5B protein,while NKILA could inhibit this effect.Furthermore,co-immunoprecipitation results showed that GALNT2 could bind to MUC5AC and MUC5B protein.RNA immunoprecipitation and RNA pull-down experiments showed that NKILA could bind to GALNT2.These evidences suggested that there are correlations among the expression of NKILA,GALNT2,MUC5AC,and MUC5B proteins in asthmatic patients.Mechanically,we concluded that NKILA can suppress the O-linked glycosylation of MUC5AC and MUC5B proteins by binding to GALNT2 and inhibit the expression of MUC5AC and MUC5B proteins.Our researches provided a potential therapeutic target for AHR.展开更多
Asthma affects about 330 million individuals worldwide while 10%of asthmatic patients develop the severe type.The two main phenotypes of severe asthma are allergic and non-allergic.Notably,remodeling and mucus o...Asthma affects about 330 million individuals worldwide while 10%of asthmatic patients develop the severe type.The two main phenotypes of severe asthma are allergic and non-allergic.Notably,remodeling and mucus overexpression are hallmarks of severe asthma upon dysregulation of MUC5AC and MUC5B.In severe allergic asthma due to the initiation of the allergic cascade,immune cells are recruited and a large number of inflammatory mediators will be produced leading to the overexpression of MUC5AC and MUC5B in the airways.Moreover,the production of mediators including tumor necrosis factorα(TNF-α)and interleukin 13(IL-13)will cause airways’muscle proliferation.Both overproductions of mucin and muscle proliferation will lead to remodeling progression.On the other hand,in severe non-allergic asthma,fewer immune cells are involved but still,the expression of MUC5AC is enhanced.However,MUC5B might increase less than the amount of its expression in allergic phenotype due to the lower number of involved immune cells and mediators.In the non-allergic phenotype mediators such as interleukin 17(IL-17)and transforming growth factorβ(TGF-β)are responsible for muscle proliferation.The result of mucus overexpression and muscle proliferation is remodeling progression in the non-allergic severe asthma.Therefore,we hypothesize that MUC5AC and MUC5B overexpression in severe allergic asthma is greater than in severe non-allergic asthma.Hence,remodeling progression is more intensive in severe allergic asthma.In conclusion,given the central roles of MUC5AC and MUC5B in mediating the asthma severity,they can be treated as potential therapeutic targets in severe asthma.展开更多
基金This work was supported by Xi’an Science and Technology Plan Project(22YXYJ0136).
文摘Objective:To explore the role of NLRP3 in mucus hypersecretion in asthmatic patients.Methods:From January 2020 to June 2022,90 patients with asthma and 60 healthy patients under the Department of Pulmonary and Critical Care Medicine of the First Affiliated Hospital of Xi’an Medical University were selected.Immunohistochemistry and enzyme-linked immunosorbent assay were performed.NLRP3 inflammasome and mucins MUC5AC and MUC5B levels in lung tissue and sputum were detected.Results:Compared to the healthy control group,the asthma group had significantly higher sputum MUC5A(20.12±5.07 versus 36.21±6.13)and NLRP3(72.31±15.13 versus 119.21±31.21)levels(P<0.05)but lower MUC5B levels(1.35±0.12 versus 0.53±0.11,P<0.05).Immunohistochemistry showed that NLRP3,MUC5AC,and MUC5B expressions were consistent with the sputum results.Conclusion:NLRP3 and MUC5AC levels are significantly increased in asthmatic patients,whereas MUC5B levels are reduced in these patients.They can be used as targets for the diagnosis and treatment of asthma.
文摘In the human airway, the gelforming mucin subtypes MUC5B and MUC5AC play important roles in biophylaxis. However, the regulation of MUC5B production is less clear than that of MUC5AC. Therefore, the regulation of MUC5B production by cell attachment and Akt was investigated in human lung epithelial NCI-H292 cells. We found that low cell attachment to culture plates induced the upregulated production of both MUC5B and MUC5AC. Cell attachment induces the activation of Akt, a serine/threonine kinase. Cell treatment with Akt inhibitor I decreased Akt phosphorylation and activation. However, MUC5B production was unaffected by Akt inhibition, whereas MUC5AC production was upregulated. MUC5B production was also unaffected by Akt inhibition in cells cultured on type IV collagen or fibronectin. These results suggest that the production of both MUC5B and MUC5AC is regulated by cell attachment. However, the regulation of MUC5B is unaffected by Akt inhibition, in contrast to that of MUC5AC.
文摘Glycosylation of mucins mediated by N-acetylgalactosaminyltransferases(GALNTs)is closely related to respiratory diseases such as asthma and chronic obstructive pulmonary disease(COPD).In addition,long non-coding RNAs(LncRNAs)participate in physiological and pathological processes through various epigenetic mechanisms.In this study,we found that a novel LncRNA named NKILA combined with multiple mucins and GALNTs potentially by several bioinformatics methods,and we used quantitative real-time PCR(RT-qPCR)to detect the expressions of NKILA,MUC5AC,MUC5B,and GALNT2 mRNA in 50 cases of asthma samples and 19 cases of normal samples,whose results showed that the expression of NKILA was significantly decreased in asthmatic samples,negatively correlated with the severity of asthma and the expressions of MUC5AC and MUC5B,while GALNT2 was significantly increased in asthmatic tissues,and positively correlated with the severity of asthma and the expressions of MUC5AC and MUC5B.In vitro,we used transient transfection technology to overexpress or interfere with NKILA and GALNT2 and then detected the expressions of MUC5AC and MUC5B via RT-qPCR and Western blot,which demonstrated GALNT2 can promote the expressions of MUC5AC and MUC5B protein,while NKILA could inhibit this effect.Furthermore,co-immunoprecipitation results showed that GALNT2 could bind to MUC5AC and MUC5B protein.RNA immunoprecipitation and RNA pull-down experiments showed that NKILA could bind to GALNT2.These evidences suggested that there are correlations among the expression of NKILA,GALNT2,MUC5AC,and MUC5B proteins in asthmatic patients.Mechanically,we concluded that NKILA can suppress the O-linked glycosylation of MUC5AC and MUC5B proteins by binding to GALNT2 and inhibit the expression of MUC5AC and MUC5B proteins.Our researches provided a potential therapeutic target for AHR.
文摘Asthma affects about 330 million individuals worldwide while 10%of asthmatic patients develop the severe type.The two main phenotypes of severe asthma are allergic and non-allergic.Notably,remodeling and mucus overexpression are hallmarks of severe asthma upon dysregulation of MUC5AC and MUC5B.In severe allergic asthma due to the initiation of the allergic cascade,immune cells are recruited and a large number of inflammatory mediators will be produced leading to the overexpression of MUC5AC and MUC5B in the airways.Moreover,the production of mediators including tumor necrosis factorα(TNF-α)and interleukin 13(IL-13)will cause airways’muscle proliferation.Both overproductions of mucin and muscle proliferation will lead to remodeling progression.On the other hand,in severe non-allergic asthma,fewer immune cells are involved but still,the expression of MUC5AC is enhanced.However,MUC5B might increase less than the amount of its expression in allergic phenotype due to the lower number of involved immune cells and mediators.In the non-allergic phenotype mediators such as interleukin 17(IL-17)and transforming growth factorβ(TGF-β)are responsible for muscle proliferation.The result of mucus overexpression and muscle proliferation is remodeling progression in the non-allergic severe asthma.Therefore,we hypothesize that MUC5AC and MUC5B overexpression in severe allergic asthma is greater than in severe non-allergic asthma.Hence,remodeling progression is more intensive in severe allergic asthma.In conclusion,given the central roles of MUC5AC and MUC5B in mediating the asthma severity,they can be treated as potential therapeutic targets in severe asthma.
