Urinary Proteome Analysis of Global Cerebral Ischemia–Reperfusion Injury Rat Model via Data-Independent Acquisition and Parallel Reaction Monitoring Proteomics

Abstract

Cerebral ischemia–reperfusion (I/R) injury is the leading cause of death in severe hypotension caused by cardiac arrest, drowning, and excessive blood loss. Urine can sensitively reflect pathophysiological changes in the brain even at an early stage. In this study, a rat model of global cerebral I/R injury was established via Pulsinelli’s four-vessel occlusion (4-VO) method. Overall, 164 urinary proteins significantly changed in the 4-VO rat urine samples compared to the control samples by data-independent acquisition (DIA) proteomics technique (1.5-fold change, p < 0.05). Gene Ontology annotation showed that the acute-phase response, the ERK1 and ERK2 cascade, endopeptidase activity, blood coagulation, and angiogenesis were overrepresented. After parallel reaction monitoring (PRM) validation, 15 differential proteins having human orthologs were verified as the potential urinary markers associated with cerebral I/R injury. Of these potential biomarkers, 8 proteins were reported to be closely associated with cerebral I/R injury. Nine differential proteins changed even when there were no clinical manifestations or histopathological cerebral damage, including FGG, COMP, TFF2, HG2A, KNG1, CATZ, PTGDS, PRVA, and HEPC. These 9 proteins are potential biomarkers for early screening of cerebral I/R injury to prevent the development of cerebral injury. KNG1, CATZ, PTGDS, PRVA, and HEPC showed an overall trend of upregulation or downregulation at 12 and 48 h after I/R injury, reflecting the progression of cerebral I/R injury. These 5 proteins may serve as potential biomarkers for prognostic evaluation of cerebral I/R injury. These findings provide important clues to inform the monitoring of cerebral I/R injury and further the current understanding of its molecular biological mechanisms.

Integrative proteomic profiling of lung tissues and blood in acute respiratory distress syndrome

“…To further evaluate the clinical value of the six target DEPs, we generated receiver operating characteristic (ROC) curves based on the aforementioned proteomics data from the plasma of ARDS patients with COVID-19 (including 5 nonsurvivors, 7 severe cases, 10 mild cases, and 8 healthy subjects) ( 10 ). The AUC values of six proteins we proposed as potential biomarkers, are ranging from 0.660 to 1.000, with more than half (5 proteins) showing an AUC greater than 0.8 ( Figures 6A-F ).…”

Proteome profiling of hippocampus reveals the neuroprotective effect of mild hypothermia on global cerebral ischemia–reperfusion injury in rats

“…An Orbitrap Fusion Lumos Tribrid mass spectrometer coupled with an EASY-nLC 1000 HPLC system (Thermo Scientific, Germany) was used to analyze the peptide sample, as previously described 15 , 16 . In detail, the digested peptides were loaded onto a trap column (75 µm × 2 cm, 3 µm, C18, 100 A°), and then transferred to a reversed-phase analytical column (50 µm × 250 mm, 2 µm, C18, 100 A°).…”

Proteome profiling of hippocampus reveals the neuroprotective effect of mild hypothermia on global cerebral ischemia-reperfusion injury in rats

“…An Orbitrap Fusion Lumos Tribrid mass spectrometer coupled with an EASY-nLC 1000 HPLC system (Thermo Scienti c, Germany) was used to analysis the pepetide sample, as previously described [15,16]. In detail, the digested peptides were loaded onto a trap column (75 µm × 2 cm, 3 µm, C18, 100 A°), and then transferred to a reversed-phase analytical column (50 µm × 250 mm, 2 µm, C18, 100 A°).…”