Study Of Naotaifang-containing Serum And Plasma By Comparative Protieomics

Objective:Through comparative study of Naotaifang containing serum and plasma protein (peptide) group, we can found that Naotaifang-containing plasma differs from serum on protein expression of anticoagulation. And we can identifiy Naotaifang characteristic protein of fibrinolytic and anticoagulation.Comparative analyzing protein expression profile of Naotaifang-containing plasma and serum samples,explore the selection of blood samples which accurately reflect the chinese herbal actual situation in vivo during pharmacological studies.Method:1.20 healthy adult SD rats were randomly divided into control group,Naotaifang treatment group by weight.The group were established in accordance with the dose by oral administration for 7 days.Before surgery, only feeded rats with water but no food.After the last administration,injected with 10% chloral hydrate intraperitoneal anesthesia,and then got blood through common carotid artery, made serum and plasma samples.2.Randomly selecting 5 rats from each group,made their serum and plasma to comparatively study their proteomics.Found the different proteins among "Naotaifang-containing plasma and normal plasma","Naotaifang-containing serum and normal serum","Naotaifang-containing serum and plasma" groups by 2-DE, and used ESI-MS/MS to identifiy important differences in proteins.Screened Naotaifang characteristic protein with fibrinolytic and anticoagulant.Results:Got 20 protein maps by 2-DE,which were normal serum,normal plasma,Naotaifang-containing serum,Naotaifang-containing plasma.Used 300DPI scan,PDQuest 7.3.0 software and got 664 protein loci.Further analysis of differences in protein loci with each group are as follows:1.Comparatively analyzed the 2-DE gel pictures of Naotaifang-containing plasma and normal plasma,20 differential proteins of 5 plasma samples were identified, among which 15 types of proteins express up-regulation and 5 down-regulation.2.Comparatively analyzed the 2-DE gel pictures of Naotaifang-containing serum and normal serum,19 differential proteins of 5 serum samples were identified,among which 15 types of proteins express up-regulation and 4 down-regulation.3.Comparatively analyzed the 2-DE gel pictures of Naotaifang-containing serum and Naotaifang-containing plasma,24 differential proteins of 5 samples were identified,among which 9 types of proteins express up-regulation and 15 down-regulation.4.Comparatively analyzed the 2-DE gel pictures of normal serum and normal plasma,7 differential proteins of 5 samples were identified,among which 4 types of proteins express up-regulation and 3 down-regulation.5.Select 10 high expressed proteins to MALDI-TOF-MS between Naotaifang-containing serum and Naotaifang-containing plasma, successfully identified six proteins (namely:inter-alpha trypsin inhibitor, heavy chain 3,Group specific component, complement factor B,Receptor Complexed With A Heterodimeric Fc, complement factor B,isoform CRA-d, Transferrin).Conclusion:The protein may be related with fibrinolysis and anticoagulant,which significant changes between Naotaifang-containing serum and Naotaifang-containing plasma. These proteins are involved in angiogenesis,inflammation and other pathological regulation of physiological processes.They are of great significance about which are the effective target and signal transduction pathway of Naotaifang.