| Background and Objective:Allergic rhinitis(AR)is a non-infectious disease of the nasal mucosa,which is mainly caused by the production of a large amount of Ig E after the human body is exposed to allergens.It is one of the refractory diseases with a high prevalence in rhinology.AR is one of the signifcant factors for inducing asthma,and brings a heavy burden to human health and socioeconomics.It is necessary to clarify the pathogenesis of AR and fnd efective prevention strategies and treatments.However,the pathogenesis of Ar is not clear.Many studies have confirmed that FOXD3-AS1(NR＿121637),a Long non-coding RNA,is significantly underexpressed in Nasal Mucosa of AR patients,suggesting that FOXD3-AS1 may be involved in the regulation of Ar Development,and is a novel regulatory gene,however,the role of FOXD3-AS1 in Ar has not been reported.In this study,we investigated the role of FOXD3-AS1 in the pathogenesis of Ar,and explored the mechanism of FOXD3-AS1in Vivo and in vitro.Methods:1.Expression of FOXD3-AS1 in Nasal Mucosa of patients with AR.(1)Nasal Mucosa tissues were collected from AR patients and healthy controls.Detection of FOXD3-AS1 expression in Nasal Mucosa by q RT-PCR.(2)Flow cytometry was used to detect the number of Th2 cells.(3)The levels of IL-25,IL-4 and IL-13 were detected by ELISA.2.The effect of FOXD3-AS1 on the secretion of IL-25 in nasal mucosal epithelial cells(NECs).(1)Human NECs were cultured in vitro and treated with different concentrations of Lipopolysaccharide(LPS).The expression and secretion of IL-25 were detected by q RT-PCR and Elisa.The expression level of FOXD3-AS1 was detected by q RT-PCR.(2)The constructed FOXD3-AS1 overexpression vector and interference vector were used to transfect into NECs cells respectively and stimulated with LPS at the same time.The expression level of FOXD3-AS1 was determined by q RT-PCR method,and the efficiency of FOXD3-AS1 overexpression and interference was examined by this method.Determination of the expression and secretion level of IL-25 by q RT-PCR and ELISA,and the effect of FOXD3-AS1 on the expression and secretion of IL-25 was analyzed.3.FOXD3-AS1 inhibits Th2 cell immunity(1)By inhibiting the secretion of IL-25 in NECs.FOXD3-AS1 is transfected into NECs by over-expression vector and interfering vector respectively,and is stimulated by LPS.Supernatant was collected and incubated with CD4~+T cells.The proportion of Th2 cells in CD4~+T cells was detected by flow cytometry,and the expressions of IL-4 and IL-13 were detected by q RT-PCR.(2)The NECs transfected with FOXD3-AS1 overexpression vector were treated with IL-25 recombinant protein and incubated with CD4~+T cells.The proportion of Th2 cells in CD4~+T cells was detected by flow cytometry,and the expressions of IL-4and IL-13 were detected by q RT-PCR.(3)The NECs transfected with FOXD3-AS1 overexpression vector were treated with IL-25 neutralizing antibody and incubated with CD4~+T cells.The proportion of Th2 cells in CD4~+T cells was detected by flow cytometry,and the expressions of IL-4and IL-13 were detected by q RT-PCR.Results:1.The expression of FOXD3-AS1 in Nasal Mucosa was down-regulated,while the proportion of Th2 cells and the levels of IL-25,IL-4 and IL-13 in peripheral blood of AR patients were increased.2.Overexpression of FOXD3-AS1 inhibits the expression and secretion of IL-25in NECs.3.The proportion of IL-4,IL-13 and TH2 in CD4~+T cells was significantly increased in NECs supernatant transfected with recombinant IL-25,the ratio of NECs supernatant transfected with FOXD3-AS1 overexpression vector to IL-4,IL-13 and TH2 cells in CD4~+T cells was significantly decreased.Conclusions:FOXD3-AS1 was down-regulated in Nasal Mucosa of AR patients,while FOXD3-AS1 inhibited the expression and secretion of IL-25 in NECs,thus inhibiting Th2 type immune response in AR.Therefore,FOXD3-AS1 may be a new target for AR therapy. |
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