Study On The Expression Of Differential Proteins FGG And FGB In Patients With Premature Coronary Heart Disease And Blood Stasis Syndrome

Objective: The difference of protein expression in plasma between familial premature coronary heart disease with blood stasis syndrome group,non-blood stasis syndrome group and health control group were compared by using Isotopic tags for relative and absolute quantification.Western Blot was used to analyze the differential protein expression.The development mechanism of familial premature coronary heart disease with blood stasis syndrome was elucidated at protein level.Method: 1.From August to December 2019,101 cases of coronary heart disease were collected from the first affiliated hospital of Hunan University of traditional Chinese medicine and Jinrun Hospital of traditional Chinese medicine in Changsha.According to the diagnostic criteria,familial premature coronary heart disease with blood stasis syndrome and familial premature coronary heart disease without blood stasis syndrome were screened,in the remaining population,matched with the experimental group were selected as the normal control group.After sample collection,iTRAQ technique was used to extract protein from plasma of family patients with blood stasis syndrome of premature coronary heart disease,family patients with blood stasis syndrome of premature coronary heart disease and family patients without coronary heart disease and normal control group,after trypsin,the peptide mixture was labeled with different iTRAQ reagents,and the peptides labeled with various iTRAQ reagents were mixed in equal amounts,the differential plasma proteins of the family with premature coronary heart disease(PCHD)and blood stasis syndrome(BSS)were screened by comparative proteomics analysis.2.Western Blot(WB)was used to identify the fibrinogen gamma chain(FGG)and fibrinogen beta chain(FGB),and the effect of FGB and FGG protein expression on the development of blood stasis syndrome of premature coronary heart disease(PCHD)was analyzed.Conclusion: 1.The data were collected by liquid chromatography-tandem mass spectrometry(lc-ms/MS)and the whole proteomics was studied by iTRAQ 8labeling technique.Using Proteome Discoverer 2.2 to search the original spectrum,the qualitative and quantitative information of protein can be obtaineds.All the proteins were identified as 556.After analysis and comparison among the groups,32 proteins(up-regulated 22,down-regulated 10)were expressed in the blood stasis syndrome group of PCHD family compared with the normal group,there were 35 different expression proteins(up-regulated 8,down-regulated 27)between PCHD non-blood stasis syndrome group and PCHD non-blood stasis syndrome group,and 28 different expression proteins(up-regulated 25,down-regulated 3)between PCHD non-blood stasis syndrome group and family,there were 34 differentially expressed proteins(up-regulated14,down-regulated 20)in PCHD patients with blood stasis syndrome and PCHD patients with non-blood stasis syndrome.Gene Ontology function enrichment analysis of differential proteins,Molecular Function(MF)was composed of epidermal structure,heparin binding,extracellular matrix structure,platelet-derived growth factor binding,cell scaffold structure,mucopolysaccharide binding and proteoglycan binding;Cellular Component(CC)mainly involves fibrinogen complex,extracellular matrix,intracellular insoluble membrane,blood particles,keratin filaments and intermediate fibers;The main Biological Process(BP)involved in the keratinocyte differentiation,fibrinolysis,humoral immune response,coagulation regulation,wound healing regulation.A total of 12 pathways were involved in the classification and enrichment analysis of KEGG biopathways.The P value of the differential protein related pathway was significantly associated with Staphylococcus aureus infection(p = 0.0008),complement system(p = 0.0088),and platelet activation(p = 0.0176),many proteins are up-regulated in these pathways.The interaction of different proteins was analyzed,and many proteins were found to be interrelated.It was found that FGG and FGB were involved in complement system and platelet activation of the pathway,and the P value of the two pathways was significant,the expression of FGG and FGB were up-regulated obviously.2.Western blot showed that the expression of FGB and FGG in PCHD blood stasis syndrome group was significantly higher than that in normal group(p <0.05).This result is consistent with the trend of iTRAQ analysis.