Based On ITRAQ Technology To Explore The Correlation Between Blood Stasis Syndrome Of Premature Coronary Heart Disease And ITLN1 And CFH Differential Proteins

Objective:According to the plasma protein expression of pedigree in premature coronary heart disease(PCHD)patients with blood stasis syndrome(BSS),plasma differential proteins with significant differences were screened to explore the influence of differential proteins on the pathogenesis of pedigree in PCHD with BSS,so as to provide potential risk markers for early clinical diagnosis of the disease.Method:(1)The expression of plasma protein was detected by isobaric tags for relative and absolute quantification(iTRAQ)for the pedigree in PCHD with BSS group,the pedigree in PCHD with non-BSS group and the healthy control group,the differential plasma protein profiles of the pedigree in PCHD with BSS were obtained by comparative analysis among groups,and the GO function and KEGG analysis.(2)Western Blot(WB)was used to verify intelectin 1(ITLN1),and complement factor H(CFH)of the pedigree in PCHD with BSS,the possible physiological or pathological relationship between ITLN1 and CFH and the pedigree in PCHD with BSS was analyzed.Results:(1)iTRAQ technique showed that there were 129 differential proteins in each group,including 32 differential proteins including 22 up-regulated proteins and 10 downregulated proteins in the pedigree in PCHD with BSS group compared with the healthy control group.(2)The results of GO function showed that the biological function of the differential protein between the pedigree in PCHD with BSS group and the healthy control group was related to cornification,cornified envelope and structural constituent of epidermis.(3)KEGG analysis showed that there were 20 different proteins and 13 related pathways in each group.The significant P values of different protein-related pathways in the pedigree in PCHD with BSS group and the healthy control group were Staphylococcus aureus infection(P=0.000),Complement and coagulation cascades(P=0.000),Platelet activation(P=0.001),Glycolysis/Gluconeogenesis(P=0.007),ECM-receptor interaction(P=0.011)and Protein digestion and absorption(P=0.013).The protein expression of CFH(P=0.003)and ITLN1(P=0.015)was significantly lower than that of the healthy control group according to P value,protein function,pathway and the physiological and pathological characteristics of the pedigree in PCHD with BSS.(4)The results of the WB experiment showed that ITLN1 and CFH protein expression in the pedigree in PCHD with BSS group were lower than those in the healthy control group(P<0.01),which was consistent with the results of iTRAQ technique.Conclusion:(1)There were 32 differential plasma proteins,including 10 down-regulated proteins,which suggested that the down-regulation of these proteins might be one of the mechanisms leading to the occurrence and development of PCHD with BSS.(2)Analysis of Go function and KEGG showed that the biological functions of plasma differential proteins were mostly related to keratinization,keratinization envelope and the components of epidermal structure,these pathways are related to vascular endothelial injury,thrombosis,inflammation and lipid metabolism,suggesting that these biological functions and pathways are related to the progression of PCHD with BSS.(3)The expression level of differential proteins ITLN1 and CFH were decreased(P<0.01),and they interacted with PITX3 protein.This may be because the inhibition of ITN1 and CFH leads to low expression,resulting in the decreased expression of Pitx3,affecting the development of dopaminergic neurons,and weakening the protective effect of arterial thrombosis,that causes the occurrence and development of PCHD with BSS,therefore,the low expression of ITLN1 and CFH may be a risk marker for the occurrence and development of the pedigree in PCHD with BSS.