支气管哮喘患儿肺泡灌洗液miR-98-5p及靶蛋白STAT3的表达与气道炎症的相关性分析

Correlation between expression of miR-98-5p and its target protein STAT3 in bronchoalveolar lavage fluid and airway inflammation in children with bronchial asthma

目的:探讨支气管哮喘患儿肺泡灌洗液(BALF)中miR-98-5p及靶蛋白STAT3表达水平与气道炎症的相关性。方法:选取2017年1月至2020年5月于重庆市涪陵区妇幼保健院就诊的120例支气管哮喘患儿作为哮喘组,包含72例急性发作期患儿(BA-E组)和48例缓解期患儿(BA-R组);纳入20例无支气管哮喘患儿为对照组。收集BALF,用实时荧光定量PCR法检测其中miR-98-5p相对表达量,采用酶联免疫吸附法检测STAT3和气道炎症因子(IL-6、IL-10、IL-13、TNF-α、总IgE),另外检测哮喘患儿肺功能和FeNO含量。结果:与对照组相比,哮喘患儿BALF中miR-98-5p相对表达量降低,STAT3水平升高,且BA-E组患儿miR-98-5p和STAT3水平变化更明显(P<0.05)。经TargetScan在线软件和双荧光素酶报告基因分析显示,miR-98-5p存在与STAT3 mRNA相结合的互补序列。与BA-R组相比,BA-E组FEV1、FEV1/FVC以及BALF中IL-10水平降低(P<0.001),同时FeNO和BALF中IL-6、IL-13、TNF-α、IgE水平均增高(均P<0.05)。经Pearson分析,哮喘患儿BALF中miR-98-5p与STAT3、IL-6、IL-13、TNF-α、IgE呈负相关(均r<0,P<0.001),与IL-10水平则呈正相关(r=0.288,P=0.001);而STAT3水平与IL-6、IL-13、TNF-α、IgE均呈正相关(均r>0,P<0.001),与IL-10水平则呈负相关(r=-0.285,P<0.001)。结论:支气管哮喘患儿BALF中miR-98-5p相对表达量降低,同时STAT3蛋白水平升高,且两者与气道炎症有关。

Objective:To explore the correlation between the expression levels of miR-98-5p and the target protein STAT3 in alveolar lavage fluid of children with bronchial asthma and airway inflammation.Methods:A total of 120 children with bronchial asthma who were hospitalized or outpatient in our hospital from January 2017 to May 2020 were selected into asthma group,including 72 children in acute onset(BA-E group)and 48 children in remission(BA-R group),additional 20 children without bronchial asthma into control group.The bronchoalveolar lavage fluid samples(BALF)were collected and detected for miR-98-5p,STAT3 and airway inflammatory factors(IL-6,IL-10,IL-13,TNF-α,IgE).The lung function and the content of FeNO were also detected.Results:Compared with the control group,the relative expression of miR-98-5p in the BALF samples of children with asthma in the two sub-groups decreased,and the level of STAT3 increased,and the changes in the levels of miR-98-5p and STAT3 in the BA-E group were more obvious(P<0.05).TargetScan online software and dual luciferase reporter gene analysis proved that miR-98-5p has a complementary sequence binding to STAT3 mRNA.Compared with children in the BA-R group,the levels of IL-10 in the FEV1,FEV1/FVC and BALF samples in the BA-E group decreased,while the levels of IL-6,IL-13,TNF-α,and IgE in FeNO and BALF increased(all P<0.001).According to Pearson analysis,miR-98-5p was negatively correlated with STAT3,IL-6,IL-13,TNF-α,and IgE(all r<0,P<0.001).There was a positive correlation with IL-10 level(r=0.288,P=0.001);while the STAT3 level in BALF was positively correlated with IL-6,IL-13,TNF-α,and IgE(all r>0,P<0.001),and was negatively correlated with IL-10 level(r=-0.285,P<0.001).Conclusion:The relative expression of miR-98-5p in BALF of asthmatic children decreases,while the level of STAT3 protein increases,which is related to airway inflammation.