Screening Of Serum MiRNA Differentially Expressed Profiles In Children With Asthma By Second Generation Sequencing

Objective:Asthma in children is a chronic airway inflammatory reaction,and its pathogenesis has not been fully elucidated.Due to the limitations of lung function detection technology in young children and the lack of a unified judgment standard,it is still difficult to play the role of diagnosis in many cases.As a noninvasive biological indicator of regulating immune function,miRNA has important clinical value in the diagnosis and evaluation of asthma.This study selected children with asthma for research.Just in order to find out the miRNA related to the occurrence and development of childhood asthma,and that would provide theoretical basis for the early diagnosis and treatment of childhood asthma.Method:(1)Serum samples wre collected from 3 asthmatic children and 3healthy children.(2)The second generation sequencing technology was used to analyze the differentially expressed miRNA in serum of children with asthma,and the differentially expressed miRNA in serum were screened by bioinformatics.(3)Serum samples were collected from 50 asthmatic children and 25 healthy children,and key miRNA were verified by RT-q PCR.(4)ROC curve was used to analyze the specificity and sensitivity of miRNA for clinical diagnosis of childhood asthma.Result:(1)Compared with the healthy control group,there were 18 up-regulated miRNA and 16 down-regulated miRNA in serum of children with asthma.Among them,the up-regulated miRNA expressed in all the sequenced samples with the largest differential multiple was miR-133a-3p,and the down-regulated miRNA was miR-939-5p.(2)The results showed that 5553 target genes were up regulated and 4604 were down regulated by 10 miRNA with the largest differential multiple detected.(3)miR-133a-3p regulates estrogen receptor signaling pathway and choline metabolism signaling pathway by targeting SP1 and EGFR;miR-939-5p regulates cell adhesion molecule signaling pathway by targeting NECTIN1 and NFASC.(4)The mean expression of miR-133a-3p in 50 children with asthma was6.00±4.73,and 2.26±1.49 in 25 control groups,and the difference was significant(P<0.01);the mean expression level of miR-939-5p in children asthma patients was 0.24±0.22,and that of the control group was 0.71±0.53,and the difference was significant(P<0.01).The expression trend of the two miRNA were consistent with the results of the second generation sequencing.(5)The sensitivity and specificity of miR-133a-3p were analyzed by ROC curve.the area under ROC curve(AUC)was 0.901,the optimal threshold of miR-133a-3p was 3.128,the sensitivity was 80.0%,the specificity was 88.0%,and the Youden index was 0.680.(6)The sensitivity and specificity of miR-939-5p were analyzed by ROC curve.AUC was 0.836,the optimal threshold of miR-939-5p was 0.316,the sensitivity was 80.0%,and the specificity was78.0%.The Youden index is 0.580.(7)The specificity and sensitivity of the combined diagnosis of miR-133a-3p and miR-939-5p were analyzed by ROC curve.The results showed that AUC was0.923,the sensitivity of diagnosis was 98.0%,the specificity of diagnosis was72.0%,and the Youden index was 0.700.Conclusion:(1)The differentially expressed miRNA in serum of asthmatic children were obtained.Among them,the up-regulated miRNA expressed in all the sequenced samples with the largest differential multiple was miR-133a-3p,and the down-regulated miRNA was miR-939-5p.(2)MiR-133a-3p and miR-939-5p can be used as molecular markers in serum of children with asthma for screening and diagnosis of childhood asthma,and the sensitivity and specificity of the combination of miR-133a-3p and miR-939-5p are better than that of single indicator detection.