Dynamic protein changes in the perihaemorrhagic zone of Surgically Treated Intracerebral Haemorrhage Patients

Abstract

The secondary injury cascades exacerbating the initial brain injury following intracerebral haemorrhage (ICH) are incompletely understood. We used dual microdialysis (MD) catheters placed in the perihaemorrhagic zone (PHZ) and in seemingly normal cortex (SNX) at time of surgical ICH evacuation in ten patients (range 26–70 years). Routine interstitial MD markers (including glucose and the lactate/pyruvate ratio) were analysed and remaining microdialysate was analysed by two-dimensional gel electrophoresis (2-DE) and nano-liquid chromatography tandem mass spectrometry (nLC-MS/MS). Two time intervals were analysed; median 2–10 hours post-surgery (time A) and median 68–76 hours post-ICH onset (time B). Using 2-DE, we quantified 232 ± 31 different protein spots. Two proteins differed between the MD catheters at time A, and 12 proteins at time B (p < 0.05). Thirteen proteins were significantly altered between time A and time B in the SNX and seven proteins in the PHZ, respectively. Using nLC-MS/MS ca 800 proteins were identified out of which 76 were present in all samples. At time A one protein was upregulated and two downregulated, and at time B, seven proteins were upregulated, and four downregulated in the PHZ compared to the SNX. Microdialysis-based proteomics is feasible for study of secondary injury mechanisms and discovery of biomarkers after ICH.

Proteomic investigation of protein adsorption to cerebral microdialysis membranes in surgically treated intracerebral hemorrhage patients - a pilot study

“…In a recent study of ICH patients, using a similar study design we found a difference in proteins expressed in the perihemorrhagic zone (PHZ) compared to seemingly normal cortex (SNX) [13]. In light of the results of this present study it is likely that several proteins evaded detection by adsorbing to the CMD membranes.…”

“…The proteomic analysis of the microdialysate of the three patients has been previously published [13] along with a detailed description of the sample preparation. Briefly, 40 μL of each sample was applied onto an Albumin & IgG Depletion column (GE Healthcare, Uppsala, Sweden), before being desalted and lyophilized.…”

“…Spots of interest were excised, 68 spots from membrane gel and 62 from the CSF-gel, destained, digested by trypsin and analyzed by mass spectrometry. Results of identified spots on the dialysate gels have been previously published [13]. Some of these results are included in Table 2 of this current study for clarity.…”

A proteomic investigation of protein adsorption to cerebral microdialysis membranes in surgically treated intracerebral hemorrhage patients

“…Microdialysate was collected at a time corresponding to 72 hours after ICH onset. The results of 2-DE of this microdialysate has been previously published (13).…”

“…The proteomic analysis of the microdialysate of the three patients has been previously published (13) along with a detailed description of the sample preparation. Briefly, 40 µL of each sample was applied onto an Albumin & IgG Depletion column (GE Healthcare, Uppsala, Sweden), before being desalted and lyophilized.…”