Analysis Of Expression Profiles Differences In Plasma Exosomal MicroRNA In Acute Kawasaki Disease And Its Clinical Value

Objectives:Through the differential analysis of plasma exosome miRNA in the acute phase of Kawasaki disease(KD),the expression profiles were understood,and plasma exosome miRNA associated with coronary artery lesions(CAL)were further screened and their clinical significance was discussed.Through the determination of cytokines(TGF-β1 and TNF-α)in the acute phase of KD,the changes in KD were understood,and its correlation with miRNA expression level was analyzed to further explore the pathogenesis of KD.Methods:(1)According to the enrollment criteria,10 children in the acute phase of KD and 5 children with healthy physical examination were selected for the first time in Jiangxi Provincial Children’s Hospital,peripheral blood was drawn before treatment,total plasma exosome RNA was extracted,and high-throughput sequencing technology was used to screen plasma exosome miRNA that were differentially expressed in the acute phase of KD;and searched databases for plasma exosome miRNA that were differentially expressed in the acute phase of KD.(2)40 children and 20 healthy children in the acute phase of KD were selected,and the above results were verified by RT-qPCR,and the differentially expressed miRNA in the acute phase of KD were screened.(3)40 children with KD complicated CAL were selected as CAL group,and 40 children with KD without CAL were selected as NCAL group,and the differences between groups in the relative expression level of plasma exosome miRNA were analyzed,and the miRNA with differential expression of CAL was further screened,and a risk score model for predicting CAL was established,and the predictive efficacy of this model on CAL was analyzed by ROC curve.(4)40 children and 20 healthy children in the acute stage of KD were selected,and the expression differences of TGF-β1 and TNF-α in the acute stage of KD and healthy children were detected by ELISA method,and the correlation between TGF-β1 and TNF-α and the expression level of candidate miRNA was statistically analyzed.Results:(1)The sequencing results showed that compared with the healthy children’s group,there were 34 differentially expressed miRNA in plasma exosomes in the KD group;The results of literature search showed that miR-1246,miR-210-5P and let-7i-3p were expressed differently in the acute phase of KD(let-7i-3p was consistent with the sequencing results).(2)RT-qPCR verification was performed on 7 miRNA in the above results according to the differential expression multiples and disease associations,and the results showed that miR-193a-5p,miR-203a-3p,miR-210-5p,miR-224-5p,let-7f-5p were differentially expressed in KD(P<0.05),and the expression was downregulated in the KD group,while there was no significant expression difference between miR-1246 and let-7i-3p(P>0.05).(3)The difference analysis between KD concurrent CAL and non-CAL groups showed that miR-210-5p was expressed differently in the CAL group(P<0.05)and was relatively upregulated in the CAL group,while miR-193a-5p,miR-203a-3p,miR-224-5p,and let-7f-5p had no significant expression difference between the two groups(P >0.05).The ROC curve shows that the area under the curve of miR-210-5p in the prediction model of KD concurrent CAL is 0.634(CI95%:0.511-0.757).(4)The cytokine concentration measurement results showed that TGF-β1 and TNF-α were expressed differently in the KD group and the healthy group,and the expression was upregulated in the KD group;The statistical analysis results showed that TGF-β1 had negative correlations with miR-193a-5p,miR-203a-3p,miR-210-5p and miR-224-5p,TNF-α had negative correlations with miR-193a-5p,miR-203a-3p and miR-224-5p,among which miR-210-5p was moderately negatively correlated with TGF-β1.Conclusions:(1)There are a large number of plasma exosome miRNA expressed differently between children with acute KD and healthy children.(2)Plasma exosomes miR-193a-5p,miR-203a-3p,miR-210-5p,miR-224-5p,let-7f-5p can be used as potential biomarkers for the diagnosis of KD in the acute stage.(3)In KD,plasma exosome miR-210-5p is associated with CAL and may be used as a predictor of CAL in the future.(4)TGF-β1 and TNF-α may be involved in the pathogenesis of KD and may be related to the regulation of miRNA.