Predictive Value And Potential Protective Effect Of Peripheral Blood Fibronectin In Kawasaki Disease Coronary Artery Aneurysm

BackgroundKawasaki disease(KD)is an acute systemic vasculitis syndrome commonly found in children,which can lead to long-term sequelae of coronary artery injury,such as coronary artery dilation(CAD)and coronary artery aneurysm(CAA).KD patients have been found to have persistent systemic vascular endothelial dysfunction,which may be one of the risk factors for early onset of cardiovascular disease in adulthood,thus bringing huge economic burden to patients.Therefore,it is of great clinical significance to explore new biomarkers to predict KD coronary artery injury(CAL)events.In this study,bioinformatics data mining showed that fibronectin(FN1)may be related to the occurrence of KD coronary artery injury.In consequence,the clinical relationship between the expression level of FN1 and KD was evaluated through clinical samples and in vitro cell experiments to explore the potential value of FN1 as a biomarker for predicting KD coronary artery injury.MethodsIn the first part,GSE data sets related to KD and coronary artery disease were screened from GEO database for obtaining differentially expressed genes(including FN1).To clarify the changing characteristics of FN1 in the course of KD and poor prognosis of coronary artery,we collected plasma from 96 healthy children and 263 patients diagnosed with KD in our hospital.KD patients were divided into different groups according to the course of disease and different coronary outcomes.FN1 levels in peripheral blood(p FN1+ c FN1)were determined by enzyme-linked immunosorbent assay,then the differences of FN1 in KD disease course and coronary prognosis were analyzed.In the second part,the primary human umbilical vein endothelial cells(HUVEC)were extracted from the umbilical cord,and the inflammation damage cell model was constructed by stimulating HUVEC with the cell wall component of Lactobacillus casei(LCWE)to simulate the status of coronary endothelial injury in KD patients.Different concentrations of exogenous FN1 were added to interfere with LCWE-HUVEC.CCK-8 was used to detect the cell proliferation activity and Transwell chamber was used to detect the cell migration ability,so as to evaluate the effect of exogenous FN1 on the proliferation and migration ability of HUVEC with inflammatory injury.Results1.The peripheral blood FN1 level in KD group was higher than that in the control group during the acute and recovery periods,and the difference was statistically significant(P<0.05).2.In KD group,the peripheral blood FN1 level in the acute phase was higher than that in the recovery phase,with a statistically significant difference(P<0.05).3.Compared with NCAL patients,there was no statistical difference in peripheral blood FN1 level of CAD patients in the acute phase(P>0.05),while that of CAA patients was significantly lower(P<0.05).4.There was no significant difference in peripheral blood FN1 level among KD patients with different coronary outcomes during convalescence(P>0.05).5.The peripheral blood FN1 level in acute phase was used to predict the occurrence of KD coronary artery aneurysm event,and the ROC curve showed that AUC was 0.7301(95% CI: 0.6480-0.8121,P<0.05).According to Youden index,the cut-off value was 216.1 mg/L,and its sensitivity and specificity were67.27% and 71.17%,respectively.6.The results of 48 h CCK-8 proliferation experiment showed that OD value in FN1 group was significantly higher than that in control group.In addition,the increase degree of OD value in 310mg/L FN1 group(similar to the average concentration of patients with NCAL in the acute phase)was significantly higher than that in 210mg/L FN1 group(similar to the average concentration of patients with CAA in the acute phase)(P<0.05).7.The results of cell migration experiment showed that,compared with the blank control group,the number of LCWE-HUVEC in 310mg/L FN1 group that migrated from the upper chamber to the lower chamber increased(P<0.05).However,there was no significant difference in the number of cell migration in210mg/L FN1 group(P>0.05).Conclusions1.The FN1 level in peripheral blood lower than 216.1mg/L in the acute stage may be one of the risk factors for the formation of CAA.2.Higher levels of FN1 had a stronger effect on promoting endothelial cell proliferation and migration in the KD acute phase,allowing for faster repair of damaged areas of the endothelial barrier,thereby maintaining its integrity.