Effect And Mechanism Of LncRNA In Goblet Cell Metaplasia On Asthma

BackgroundAsthma is a chronic inflammatory disease of airway,which is involved by many kinds of cells and cytokines.Studies have shown that long non-coding RNAs(lncRNAs)participate in the physiological and pathological process through regulating epigenetic modification,protein degradation,RNA splicing and other mechanisms,and play a significant role in the emergence and development of a variety of tumors,immune system diseases and so on.LncRNAs also play a role in the proliferation and apoptosis regulation of airway smooth muscle cells,glucocorticoid resistance,immune balance,etc.But there are few reports on the study of goblet cell metaplasia.MethodsIn this study,fluorescence in situ hybridization(FISH)and q PCR were used to detect changes in the expression level of NR2F2-AS1 in bronchoalveolar lavage fluid(BALF)and mice lung tissues.Then by over-expression of NR2F2-AS1 in 16 HBE cells,we detected the expression of MUC5 AC which was an important marker of goblet cell metaplasia.In vivo,the mice model of asthma were treated AAV virus expressing NR2F2-AS1 through intratracheal administration,then Periodic Acid-Schiff stain and pulmonary function test were used to evaluate the effects.At the same time,by FISH technology we found NR2F2-AS1 was located in the cytoplasm,and its possible regulatory mechanism was further explored by methods such as dual-luciferaseexperiment,q PCR,and western blot.Results(1)LncRNAs were involved in the regulation of asthmatic airway epithelium goblet cell metaplasia: Sequencing analysis revealed that there were many differences on the expression profile of lncRNAs in BALF between asthmatic patients and foreign body group.At the same time,IL-13,an important inducer of goblet cell metaplasia,could regulate the expression level of some lncRNAs.Overexpression of NR2F2-AS1 in 16HBE cells could alleviate MUC5 AC level.(2)NR2F2-AS1 decreased in asthma group: FISH and q PCR analysis of mice lung tissue found that the expression of NR2F2-AS1 in the asthma group decreased consistent with sequencing results.(3)Overexpression of NR2F2-AS1 in vivo could alleviate airway epithelium goblet cell metaplasia: There were obvious effects on decreasing the goblet cell metaplasia but the inflammation level in asthma mice who were treated virus expressing NR2F2-AS1.And overexpression of NR2F2-AS1 may improve lung function in mice model of asthma.(4)Exploring the mechanism of NR2F2-AS1 on regulation of goblet cell metaplasia: Excluding that NR2F2-AS1 regulated MUC5 AC expression by as competitive endogenous RNAs(ce RNAs)and regulating NR2F2 in cis,it was a positive regulator of FOXA2 and participated in the regulation of Hippo and Wnt signaling pathways.ConclusionsLncRNAs are involved in the regulation of goblet metaplasia of airway epithelial cells in asthma,and NR2F2-AS1 inhibits goblet cell metaplasia.Enchanced expression of NR2F2-AS1 in vivo can alleviate the accumulation of glycogen in airway epithelium and improve lung function.