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
目的基于蛋白质组学技术筛选正常组、疏肝调神针刺组月经性偏头痛患者血清差异表达蛋白,为疏肝调神针法治疗月经性偏头痛患者提供参考依据。方法收集6例女性健康体检者血清(A组),6例月经性偏头痛患者为疏肝调神针刺组(B组),并在治疗前...目的基于蛋白质组学技术筛选正常组、疏肝调神针刺组月经性偏头痛患者血清差异表达蛋白,为疏肝调神针法治疗月经性偏头痛患者提供参考依据。方法收集6例女性健康体检者血清(A组),6例月经性偏头痛患者为疏肝调神针刺组(B组),并在治疗前后分别取血,治疗前为Bq组,治疗后为Bh组。按标号最终得到18个混样。通过蛋白提取、酶切、液相色谱-质谱串联分析、生物信息分析等对样本进行定量蛋白组的研究。对差异表达蛋白进行GO富集、Reactome通路分析以及蛋白互作网络分析。结果Bq/Bh:上调蛋白10个,下调蛋白7个;A/Bh:上调蛋白18个,下调蛋白51个。GO富集分析显示,A/Bh中,最显著富集的20个亚类中生物学过程有14个亚类,细胞元件有2个亚类,分子功能有4个亚类。Bq/Bh中,最显著富集的20个亚类中生物学过程有12个亚类,细胞元件有6个亚类,分子功能有2个亚类。Reactome通路富集分析结果显示:Bq与Bh相比,O-linked glycosylation of mucins(R-HAS-913709)通路和Cellular response to chemical stress(R-HAS-9711123)通路在富集程度及差异蛋白的数量上最为显著。蛋白互作网络分析显示:A/Bh中FN1(fibronectin-1)、UBB(Polyubiquitin-B)、TXN(Thioredoxin)等在月经性偏头痛患者的血液中表达下调,DPP4(Recombinant human dipeptidyl peptidase 4)、SLPI、DNPEP(aspartyl aminopeptidase)等上调。结论疏肝调神针法治疗月经性偏头痛可能通过调控黏蛋白的O-糖基化通路及O型糖基化转移酶GALNT2的表达发挥作用。展开更多
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.
文摘目的基于蛋白质组学技术筛选正常组、疏肝调神针刺组月经性偏头痛患者血清差异表达蛋白,为疏肝调神针法治疗月经性偏头痛患者提供参考依据。方法收集6例女性健康体检者血清(A组),6例月经性偏头痛患者为疏肝调神针刺组(B组),并在治疗前后分别取血,治疗前为Bq组,治疗后为Bh组。按标号最终得到18个混样。通过蛋白提取、酶切、液相色谱-质谱串联分析、生物信息分析等对样本进行定量蛋白组的研究。对差异表达蛋白进行GO富集、Reactome通路分析以及蛋白互作网络分析。结果Bq/Bh:上调蛋白10个,下调蛋白7个;A/Bh:上调蛋白18个,下调蛋白51个。GO富集分析显示,A/Bh中,最显著富集的20个亚类中生物学过程有14个亚类,细胞元件有2个亚类,分子功能有4个亚类。Bq/Bh中,最显著富集的20个亚类中生物学过程有12个亚类,细胞元件有6个亚类,分子功能有2个亚类。Reactome通路富集分析结果显示:Bq与Bh相比,O-linked glycosylation of mucins(R-HAS-913709)通路和Cellular response to chemical stress(R-HAS-9711123)通路在富集程度及差异蛋白的数量上最为显著。蛋白互作网络分析显示:A/Bh中FN1(fibronectin-1)、UBB(Polyubiquitin-B)、TXN(Thioredoxin)等在月经性偏头痛患者的血液中表达下调,DPP4(Recombinant human dipeptidyl peptidase 4)、SLPI、DNPEP(aspartyl aminopeptidase)等上调。结论疏肝调神针法治疗月经性偏头痛可能通过调控黏蛋白的O-糖基化通路及O型糖基化转移酶GALNT2的表达发挥作用。
