MicroRNA-466 And GPR18 MRNA Expression In Peripheral Blood And Diagnosis Value In The Children With Sepsis

ObjectiveTo explore the expression of miRNA-466 and GPR 18 m RNA in mononuclear cells of children with sepsis,to explore the correlation between miRNA-466,GPR 18mRNA and SOFA score,to explore the regulatory relationship between miRNA-466and GPR 18 mRNA,and the diagnostic value of miRNA-466 and GPR 18 mRNA in children with sepsis.MethodsFrom October 2017 to June 2019,90 children with sepsis diagnosed in The Fourth Peoples Hospital of Shenzhen were selected as sepsis group and 60 healthy children as control group.The clinical data of children in sepsis group and control group were collected,and 5 ml peripheral venous blood was collected respectively.The expression of miRNA-466 and GPR 18 mRNA in PBMC of the two groups was measured by PCR.The WBC counts,CRP concentrations,miRNA-466 and GPR 18mRNA expression levels in mononuclear cells of children between sepsis group and control group were compared.Pearson correlation analysis was used to analyze the correlation between mi RNA-466 and GPR 18 mRNA in peripheral blood mononuclear cells of children with sepsis and SOFA score.ROC curve analysis was used to compare the diagnostic value of CRP concentration,miRNA-466,GPR 18mRNA and combined detection in children with sepsis.Luciferase reporter gene experiment verified the expression relationship between miRNA-466 and GPR 18.The expression of GPR 18 in PBMC cells was detected by Western blot.Results(1)The blood WBC count of children in sepsis group was significantly higher than that of healthy control group(11.05±3.86 vs.8.53±2.04,P<0.05).The blood CRP concentration of children in sepsis group was significantly higher than that of healthy control group(38.43±11.52VS.7.5±2.07,P<0.05),.(2)The expression level of mi RNA-466 in PBMC of sepsis group was significantly higher than that of control group(1.35±0.52 vs.1.27±0.34 P<0.05).The relative expression level of GPR 18mRNA in PBMC of sepsis group was higher than that of control group(1.53±0.54 vs.1.32±0.38 P<0.05).(3)Pearson correlation showed that there was no significant correlation between WBC count and SOFA score in peripheral circulation of children with sepsis(r=0.238,P=0.543).There was a significant positive correlation between CRP concentration and SOFA score in peripheral circulation of children with sepsis(r=0.472,P<0.01).And there was a positive correlation between miRNA-466and SOFA score in peripheral blood mononuclear cells of children with sepsis(r=0.489,P<0.001).There was a positive correlation between GPR 18 mRNA and SOFA score in peripheral blood mononuclear cells of children with sepsis(r=0.507,P<0.001).There was a positive correlation between miRNA-466 and GPR 18 mRNA expression in peripheral blood mononuclear cells of children with sepsis(r=0.618,P<0.001).(4)When the cutoff value is 33.7,the ROC~aucuc of blood CRP concentration was 0.758,95%CI was 0.669-0.817,sensitivity is 68.0%,specificity was 71.3%.When the cutoff value is 1.32,the ROCauc of miRNA-466 in peripheral blood mononuclear cells is 0.901,95%CI was 0.813-0.969,sensitivity was 84.5%,specificity was 87.9%.when the cutoff value was 1.38,the ROC~aucuc of GPR 18 in peripheral blood mononuclear cells was 0.858,95%CI was 0.761-0.917,sensitivity was 71.0%,specificity was 92.3%.When combined detection of the relative expression of miRNA-466 and GPR 18 mRNA in mononuclear cells,the ROC~aucuc was 0.947,95%CI was 0.898-0.981,sensitivity was 90.9%,specificity was 93.9%.(5)MicroRNA.org,the on-line target gene prediction website showed that there was a miRNA-466 target binding site in the 3’-UTR of GPR 18 mRNA.The expression of miRNA-466 mimic significantly decreased the activity of luciferase(P<0.05).After transfection of miRNA-466 mimic in PBMC cells,the expression level of miRNA-466 increased significantly.After transfection of miRNA-466 inhibitor,the expression level of miRNA-466 decreased significantly.Western blotting showed that the expression of GPR 18 protein increased and decreased after up-regulation and down-regulation of miRNA-466 respectively.ConclusionsThe blood CRP concentrations and miRNA-466 expression levels,GPR 18mRNA expression levels in peripheral blood mononuclear cells were positively correlated with SOFA score in the children with sepsis.There was a positive correlation between mi RNA-466 and GPR 18 mRNA expression in peripheral blood mononuclear cells.Combined detection of mi RNA-466 and GPR 18 mRNA can be helpful for diagnosing sepsis in children.There are targeted regulation relationship between miRNA-466 and GPR 18.