| Congenital heart disease, which is a major cause of infantile death exceptinfectious disease, is an important component of pediatric cardiovasculardisease and constitutes a major percentage of clinically significant birthdefects. Ventricular septal defect(VSD), which accounts for about25to50per100congenital heart disease, is the main type of birth defects. The etiologyand pathogenesis of VSD may involve lots of multiple factors, multiple genesand multiple pathways, which plays an important role in heart development.Previous studies,researched the pathogenesis of VSD, focused onenvironmental risk factors, genetic, and so on. But studying the environmentalrisk factors and the susceptibility gene mapping is not enough to understand ofthe mechanism of VSD completely, we must find out the proteins which areclosely related to the abnormal differentiations and developments in theprocess of cardiac morphogenesis and study their functions. Proteomicsresearch, coupled with a series of new technologies, has developed rapidlynew capability as an important branch of life science research. In ourpreliminary study,26differentially expressed proteins related with VSD wereidentified, in which13proteins were over-expressed (>1.5fold) and8proteinswere under-expressed (<0.67fold) in VSD group, compared with ASD group, TOF group and healthy group, by using isobaric tags for relative andabsolute quantitation (iTRAQ) combined with LC-MALDI-TOF/TOFtechnique. According to the appraisal to peptides matching and confidence,Fibonectin(FN) probably can be used as a serum candidate biomarker for VSD.In this experiment we used Western blot and enzyme linked immunosorbentassay to validate whether fibronectin can be used as the serum molecularmarker of Ventricular septal defect, and then analyzed fibronectin by using thebioinformatics tools and methods, which provided the foundations to discoverthe pathogenesis of VSD and new insights into potential causes of CHD.Objective:To validate whether Fibronectin can be used as the serummolecular marker of Ventricular septal defect and to analysize fibronectin byusing the bioinformatics tools and methods. Methods:1. serum samples wereobtained from20children with VSD,20children with atrial septaldefect(ASD),20children with tetralogy of Fallot (TOF) and20healthychildren, who were well matched in age, gender and race, respectively.Fibronectin expression level in serum samples was measured by Western blotand enzyme linked immunosorbent assay.2. The bioinformatics was used toanalysis FN’s basic physical and chemical properties, structure, function,metabolic pathway, the network control et al. Results:1. Western blot analysisshowed that Fibronectin was detected in all serum samples of VSD, ASD,TOF and healthy children. However, the Fibronectin expression level in VSDgroup was significantly higher than the other three groups (p<0.05).2. TheELISA indicated that the concentration of fibronectin in serum among thepatients of VSD group was (67.08±27.67)ug·L-1, the ASD group was(37.65±15.07)ug·L-1, the TOF group was (28.08±11.24)ug·L-1, and thehealthy group was (31.15±11.18)ug·L-1. The difference of concentration among the four groups is significant(p<0.05); Further comparison betweenevery two groups, the concentration of VSD patients is difference from that ofthe other three groups respectively(p<0.05), and the difference ofconcentration among the other three groups was not significant(p>0.05).3.The results from bioinformatics analysis showed FN, which contained2386amino acid, MW=262624.5Da, pI=5.46, was a dimeric glycoprotein and aubiquitous and essential component of the extracellular matrix (ECM). Itplayed a major role in cell adhesion, growth, migration, and differentiation.Conclusions: The Fibronectin expression level is higher in VSD patien’s erum.Fibronectin, which plays a major role in cell adhesion, growth, migration, anddifferentiation, can be used as a serum biomarker for VSD and need to befurther validated in large clinical samples study. |
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