Study On The Roles Of LncRNA AK085865 In Regulationing Of Macrophage Polarization In Allergic Asthma

Objective: In this project,we intend to explore the effects of lncRNA AK085865 on airway inflammation,airway remodeling,and airway hyperresponsiveness in a mouse model of allergic asthma.we further explore the effects of AK085865 regulating macrophage polarization in the in vivo environment on allergic asthma mice.we aim to elucidate the role of AK085865 in allergic asthma mouse model and its cellular immunological mechanism,thus providing a new theoretical basis and experimental basis for the study of pathogenesis and biological control of allergic asthma.Methods:(1)Grouping:female wild-type C57BL/6 mice aged 6-8 weeks were randomly divided into PBS group,WT-Df1 group and female knockout mice aged 6-8 weeks were divided into KO-Df1 group;(2)Modeling: The WT-Df1 group and KO-Df1 group were induced by dust mite protein(Dermatophagoidesfarinae1,Df1)combined with atomization attack to establish an allergic asthma model in C57BL/6 mice,while PBS group was sensitized and atomized;(3)Detection: The expression levels of alveolar lavage fluid(Bronchoalveolar lavage fluid,BALF)cells and lung tissue in asthmatic mice lncRNA AK085865 detected by fluorescence quantitative PCR;The histopathological changes of mice were observed by HE and PAS staining to compare the infiltration level of inflammatory cells around trachea and airway remodeling level in mice;The airway hyperresponsiveness of mice in each group was observed by means of lung function instrument;The number of associated inflammatory cells in airway tissue was detected by flow cytometry;The expression of inflammatory cytokines inBALF supernatant and lung tissue homogenate was detected by ELISA;The proportion of macrophages M1/M2 BALF and lung tissues was detected by flow cytometry;Macrophage-related markers were qPCR detected in BALF cells and lung tissues;The proportion and number of innate lymphocyte populations in BALF and lung tissues were detected by flow cytometry;The changes of the above indexes were observed in mice adoptive back-feeding macrophages.Results:(1)The model of asthma was constructed by Df1 sensitization atomization,and the lung histopathology of PBS group and WT-Df1 group was tested to accord with the characteristics of the model,which suggested that the model was successful;(2)The expression levels of lncRNA AK085865 in WT-Df1 group were expressed in both BALF cells and lung tissues,but higher in BALF cells group compared with the group;(3)The inflammatory response around the bronchus and the decrease of goblet cell proliferation and mucus secretion,airway hyperresponsiveness and the decrease of inflammatory cell number and inflammatory cytokine expression in BALF and lung tissue in KO-Df1 group were compared with the mice;(4)Using flow cytometry and qPCR detection,lncRNA AK085865 knockdown significantly reduced the proportion of macrophages M2 allergic airway inflammation in mice;(5)M0 macrophages derived from bone marrow of WT mice were transported back to KO-Df1 mice and it was found that the absence of lncRNA AK085865 in macrophages was the main cause of decreased asthma sensitivity;(6)The proportion of ILC2 s cells in BALF and lung tissues decreased compared with WT-Df1 mice,the level of Th2 cytokines secreted by ILC2 s cells decreased(IL-5?IL-13)and the expression of only IL-5 decreased in lung tissues.Conclusions: Lnc RNA AK085865 knockdown reduces Df1-induced susceptibility to allergic asthma in mice by reducing M2 macrophages;LncRNA AK085865 regulating susceptibility to allergic asthma may be associated with regulating M2 macrophage and ILC2 s cell differentiation.