Plasma proteomic changes during hypothermic and normothermic cardiopulmonary bypass in aortic surgeries

Abstract

Deep hypothermic circulatory arrest (DHCA) is a protective method against brain ischemia in aortic surgery. However, the possible effects of DHCA on the plasma proteins remain to be determined. In the present study, we used novel high-throughput technology to compare the plasma proteomes during DHCA (22°C) with selective cerebral perfusion (SCP, n=7) to those during normothermic cardiopulmonary bypass (CPB, n=7). Three plasma samples per patient were obtained during CPB: T1, prior to cooling; T2, during hypothermia; T3, after rewarming for the DHCA group and three corresponding points for the normothermic group. A proteomic analysis was performed using isobaric tag for relative and absolute quantification (iTRAQ) labeling tandem mass spectrometry to assess quantitative protein changes. In total, the analysis identified 262 proteins. The bioinformatics analysis revealed a significant upregulation of complement activation at T2 in normothermic CPB, which was suppressed in DHCA. These findings were confirmed by the changes of the terminal complement complex (SC5b-9) levels. At T3, however, the level of SC5b-9 showed a greater increase in DHCA compared to normothermic CPB, while 48 proteins were significantly downregulated in DHCA. The results demonstrated that DHCA and rewarming potentially exert a significant effect on the plasma proteome in patients undergoing aortic surgery.

Does the Rewarmed Heart Restore the Myocardial Proteome to That of the Pre-Cooled State? – A Proteomic Analysis of Surgical Samples –

“…40 However, we could not detect SORBS2 protein in plasma taken after reperfusion in our previous proteomic analysis. 14 The 7 isoforms of 14-3-3 proteins have multiple roles in the apoptotic signaling network and work for signaling integration. 41 14-3-3 proteins can contribute to cell survival by sequestering pro-apoptotic proteins, such as BAD, BAD and c-ABl.…”

“…13, 14 Spotted peptide samples were analyzed by a 5800 MALDI-TOF/TOF MS/MS Analyzer with The 66 identified proteins were matched to a metabolic pathway (ko 01100) and highlighted by black in the pathway map, indicating that identified proteins were intensively located on glycolysis (marked by "G"), TCA cycle (marked by "T") and oxidative phosphorylation (marked by "O") in the map. (B) TCA cycle ("T," ko 00020) map is illustrated and 13 matched proteins/associated enzyme commission numbers (EC) are highlighted in pink.…”

“…13, 14 In brief, equal amounts of the T1, T2, and T3 samples from each patient were denatured and reduced, the cysteines alkylated, and finally digested with trypsin (AB Sciex). Each digest was labeled with a different iTRAQ tag using the iTRAQ reagent 4-plex kit (AB Sciex).…”

“…Proteins satisfying these criteria were regarded as having statistical significance. 13, 14 The KEGG was used to assess whether these significant proteins could be associated with well-defined canonical or signaling pathways. PANTHER software (version 9.0, http:// www.pantherdb.org) was used to test for statistical overrepresentation of gene ontology (GO, http://www.geneontology.…”

“…13, 14 In brief, myocardial samples were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and immunoblotted using rabbit polyclonal IDH3A antibody (Proteintech, Chicago, IL, USA), mouse monoclonal gelsolin (GSN) antibody (Sigma, St. Louis, MO, USA) and anti-rabbit IR dye 680-conjugated IgG (LI-COR, Lincoln, NE, USA). Protein bands were visualized using an Odyssey (LI-COR) infrared imaging system and their intensities measured for densitometric analyses of IDHA3 and GSN.…”