The Variation Of Microparticles In Circulating Blood During Perioperative Period Of Coronary Artery Bypass Grafting

Objective:Variation of microparticles in CPB-CABG and OPCABG before and after operation were dected, including the counting of microparticles, its source and proteomic analysis. We will compare the effect of two operation methods of microparticles, effect on the microparticles before and after operation.Materials and Methods:20 patients (5 cases are female and 15 cases are male) were selected in our heart center for CABG in our hospital from March to November in 2014. The average age of the patients was 67±13 year, and the average weight was 65±12 kg. CABG inclusion criteria were serious of left main(stenosis>50%) or three vessel pathological changes(three vessel stenosis>75%).The patients were randomly divided into group A(OPCABG) and B(CPB-CABG), each group include 10 patients, include g A0(before OPCABG), Ai(after OPCABG), B0(before CPB-CABG), B,(after CPB-CABG), according to different operations. Venous blood of the patients were collected at 12h before the operation, after the operation immediately, and treatmented all samples in time. The amount of microparticles, endothelial cells MPs(EMPs), platelet MPs(PMPs)and leukocyte MPs(LMPs) were detected by flow cytometer, proteomics of microparticles was detected by mass analyzer, and thevariation of B-natriuretic peptide, C-reactive protein, myoglobin in peroperative period were also detected in this group. Data analysis:The data detected by flow cytometer were displayed as (x±s), paired t-test was used to analyze; data of proteomics collected and analyzed by Thermofisher Proteome Discoverer 1.3 and Mascot:2.3.0.Results:1.There was no significant difference in basic clinical data of patients. No statistical difference compared preoperative BNP, CRP and MB, compared with the preoperative, BNP, CRP and MB were increased after surgery, there were significant difference, compared different operation, CPB-CABG is more significant to increase their than OPCABG.2.Compared to Ao, the number of positive Annexin-V, CD 144 and CD41a antibody in microparticles of Ai were significantly increased, but there is no statistical difference in the increase of positive CD45 antibody. Compared to Bo, the number of positive Annexin-V, CD144 and CD41a antibody in microparticles of B1 were significantly increased, but there was no statistical difference in the increase of positive CD45 antibody. These results indicated that microparticles were elevated by OPCABG and CPB-CABG, especially the microparticles which derived by endothehal cells and platelets, but there is no obviously change of which derived by leukocyte. There was no significant difference compared Bo with Ao. Compared to A1, the number of positive Annexin-V, CD 144 antibody in microparticles of B1 were significantly increased, but there was no statistical difference in the increase of positive CD45 and CD41a antibody. These results indicated that microparticles was elevated more significantly by CPB-CABG than OPCABG.3.There were 201 kinds of proteins identified by iTRAQ technology in the uniprot-human library. Compared to Ao in the uniprot-human Library, there were 75 differential proteins in A1 in total, of which up-regulated proteins were 35, down regulated proteins were 40; Compared to Bo in the uniprot-human Library, there were 101 differential proteins in B1 in total, of which up-regulated proteins were 49, down-regulated proteins were 52. The results show that there were more differential proteins were detected after CPB-CABG than OPCABG. There were 48 kinds of common differentially expressed proteins in two kinds of operation, of which down-regulated proteins were 11, up-regulated proteins were 14. After coronary artery bypass grafting, the expression of fibrinogen gamma chain (P02679) and plasma vascular factor (P04275) were up-regulated which involved in platelet activation process, and the ratio of them were 1.337 and 2.336; the expression of Complement factor I (P05156) and complement C5 (P01031) were up-regulated which involved in the activation of the complement system,and the ratio of them were 2.445 and 1.450; the expression of hageman factor (P00748) and coagulation factor XIII chain (P00488) were up-regulatedwhich involved in the coagulation process, and the ratio of them werel.220 and 1.745; The expression of platelet factor-4 (P02776) and latelet basic protein (P02775)were up-regulated which involved in the inflammatory process, their ratio were 1.479 and 1.274; The expression of adiponectin (Q15848) and apolipoprotein A (P08519) were up-regulated which involved in the signal transduction expression were up-regulated, tand the ratio of them were 1.235 and 1.473. These variation in protein show that microparticles participated in the activation of platelet and complement system, blood clotting, the aggregation of inflammation and signal transduction.Conclusion:1.No matter from quantity, sources and proteomics have significant change, the levels of MPs were elevated as BNP, CRP, MB in the blood after coronary artery bypass graft, and this elevation was more obvious in CPB-CABG.2.After CABG, MPs were elevated by inflammation, coagulation disorder and other pathological process of body, so the level of MPs seem reflect the degree of damage on the body during surgical operation.3.The MPs count, differential count and proteomic analysis showed:elevate the level and change component of MPs can promote systemic inflammatory reaction, which is one of the factors of pathological injury to the body during on-pump and surgical operation, and this effect of elevation may be more obvious than the effect of MPs induced by body injury.The pathological significance of research has great necessity.4.Not only is the MPs a result of trauma during perioperation period of CABG, and also is one of the factors involved in the pathological process, MPs may become a biomarker to predict postoperative complications after on-pump operation.