Risk Factors Of Coronary Artery Damage And Intravenous Immunoglobulin Resistance And Study On The Mechanism Of NFAT2 And VDAC On Neutrophils In Kawasaki Disease

Part 1:Analysis of risk factors of coronary artery damage and intravenous immunoglobulin resistance in Kawasaki diseaseObjective:The aim of this study is to analyze the risk factors of coronary artery damage and intravenous immunoglobulin(IVIG)resistance in children with Kawasaki disease in Suzhou area.Methods:The clinical data of 276 children with Kawasaki disease,46 children with respiratory tract infection and 58 healthy children who were hospitalized in the Children’s Hospital of Soochow University in 2019 were collected.The Kawasaki disease group was divided into groups according to whether there was coronary artery damage and whether they were sensitive to intravenous immunoglobulin(IVIG).The age,gender and laboratory indexes were compared between groups,and the significant indexes were analyzed by logistic regression and ROC curve analysis to find the independent risk factors of coronary artery damage and intravenous immunoglobulin resistance.Results:1.The white blood cell count(WBC),neutrophil percentage(N%),platelet(PLT),neutrophil-to-lymphocyte ratio(NLR),platelet-to-lymphocyte ratio(PLR)of the KD group in the acute phase were all Higher than the normal control group,hemoglobin(Hb),hematocrit(HCT)was lower than the normal control group,the difference was statistically significant(P<0.05).2.The WBC,N%,PLT,NLR,C-reactive protein(CRP)and erythrocyte sedimentation rate(ESR)of the KD group in the acute phase were higher than those of the fever control group,and Hb,HCT,serum sodium and albumin(ALB)were lower than those of the fever control group,the difference was statistically significant(P<0.05).3.The WBC,N%,NLR,PLR,CRP,ALT,CD3-CD19+ of the complete KD group in the acute phase were higher than those of the incomplete KD group,and the serum sodium,albumin(ALB),total cholesterol(TCHOL),and CD3+were lower than those of the incomplete KD group.The difference was statistically significant(P<0.05).4.When the age is≤30 months,the sensitivity for diagnosis of coronary artery damage is 68.3%and the specificity is 50.3%;when the gender is boy,the sensitivity is 67.2%and the specificity is 48.1%;when ALT≥54.15U/L,the sensitivity is 42.4%,the specificity is 77.4%;when ALB≤35.95g/L,the sensitivity is 30.3%,the specificity is 87.2%;when satisfying age<30 months,male gender,ALT≥54.15U/L,ALB≤35.95g/L,the sensitivity for diagnosis of coronary artery damage was 58.0%and the specificity was 70.3%.5.When HCT≤0.316L/L,the sensitivity for diagnosis of IVIG resistance is 59.3%and the specificity is 77.5%;when the serum sodium≤132.5mmol/L,the sensitivity is 55.6%and the specificity is 85.2%;When total bilirubin≥5.45μmol/L,the sensitivity is 82.6%and the specificity is 56.2%;When satisfying HCT≤0.316L/L,serum sodium≤132.5mmol/L,and total bilirubin≥5.45μmol/L,the sensitivity for diagnosis of IVIG resistance was 55.6%,and the specificity was 95.8%.Conclusion:1.WBC,N%,PLT,CRP,ESR,NLR,and PLR in acute stage of children with Kawasaki disease increased,while Hb,HCT,and albumin decreased.2.Age(≤30 months),gender(male),ALT(≥54.15U/L),ALB(≤35.95g/L)are independent risk factors for coronary artery damage in children with Kawasaki disease.3.HCT(≤0.316L/L),serum sodium(≤132.5mmo/L),and total bilirubin(≥5.45μmol/L)are independent risk factors for IVIG resistance in children with Kawasaki disease.Part2:Expression and correlation analysis of NFAT2 and VDAC in neutrophils of Kawasaki disease.Objective:To investigate the effect and new mechanism of NFAT2 and VDAC in neutrophils in the Kawasaki disease immune vasculitis.Methods:1.Using gene chip technology to screen out differentially expressed genes in neutrophils,and using GO(Gene Ontology)to analyze genes related to mitochondrial function.2.Using bioinformatics methods to predict the target gene of NFAT2.3.Real time PCR was used to verify the mRNA expression of NFAT2 and VDAC in neutrophils in 61 children with KD,and to analyze the correlation between NFAT2 and VDAC.Results:1.The genes differentially expressed in neutrophils of Kawasaki disease and related to mitochondrial function are:VDAC3,NMT1,PPP3R1,YWHAQ,CASP8,MAPK8,HTT,BNIP3,BNIP3L,ACAA2,IER3,BAX,UCP2,BCL2L1,PAM16,DNAJC19 And COX15.2.VDAC3 may be the target gene of NFAT2 on mitochondria3.The mRNA relative expression level of NFAT2 in the KD group before treatment was lower than that in the normal control group and the fever control group,the difference was statistically significant(P<0.05).The CAL group was higher than the NCAL group,but the difference was not statistically significant.(P>0.05).4.The mRNA relative expression levels of VDAC1-3 in the KD group before treatment were significantly lower than those in the normal control group and the fever control group(P<0.05),and the CAL group was higher than the NCAL group,but there was not statistical significance(P>0.05).5.After treatment,the mRNA expression levels of NFAT2 and VDAC 1-3 in the KD group were higher than that before treatment,the difference was statistically significant(P<0.05).Compared with the normal control group and the fever control group,the difference was not statistically significant(P>0.05)6.In the KD group,NFAT2 was positively correlated with VDAC1-3,and the difference was statistically significant(P<0.05).7.In the normal control group,there was no correlation between NFAT2 and VDAC1-3,and the difference was not statistically significant(P>0.05)8.In the fever control group,NFAT2 was positively correlated with VDAC2 and VDAC3,and the difference was statistically significant(P<0.05).Conclusion:1.NFAT2 and VDAC in PMNs are involved in the immune vasculitis in the acute phase of Kawasaki disease.2.In Kawasaki disease,NFAT2 may regulate the transcription of VDAC,thereby regulating the apoptosis of neutrophils,and participate in the immune response process of Kawasaki disease.3.Mitochondria are involved in immune inflammation of Kawasaki disease,and the pathways of NFAT2 and VDAC may be potential targets for KD therapy.