phosphocreatine and Infarction--Middle-Cerebral-Artery

We performed metabolomic analyses of mouse brain using a transient middle cerebral artery occlusion (tMCAO) model with Matrix Assisted Laser Desorption/Ionization (MALDI)-mass spectrometry imaging (MSI) to reveal metabolite changes after cerebral ischemia. We selected and analyzed three metabolites, namely creatine (Cr), phosphocreatine (P-Cr), and ceramides (Cer), because these metabolites contribute to cell life and death. Eight-week-old male C57BL/6J mice were subjected to tMCAO via the intraluminal blockade of the middle cerebral artery (MCA) and reperfusion 60 min after the induction of ischemia. Each mouse was randomly assigned to one of the three groups; the groups were defined by the survival period after reperfusion: control, 1 h, and 24 h. Corrected samples were analyzed using MALDI-MSI. Results of MSI analysis showed the presence of several ionized substances and revealed spatial changes in some metabolites identified as precise substances, including Cr, P-Cr, Cer d18:1/18:0, phosphatidylcholine, L-glutamine, and L-histidine. Cr, P-Cr, and Cer d18:1/18:0 were changed after tMCAO, and P-Cr and Cer d18:1/18:0 accumulated over time in ischemic cores and surrounding areas following ischemia onset. The upregulation of P-Cr and Cer d18:1/18:0 was detected 1 h after tMCAO when no changes were evident on hematoxylin and eosin staining and immunofluorescence assay. P-Cr and Cer d18:1/18:0 can serve as neuroprotective therapies because they are biomarker candidates for cerebral ischemia.

Topics: Animals; Ceramides; Creatine; Disease Models, Animal; Infarction, Middle Cerebral Artery; Male; Metabolomics; Mice; Mice, Inbred C57BL; Phosphocreatine; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

The influence of creatine or its derivates on the cell energy potential may be one of the possibl approaches to induce neuroprotection. Effect of creamide (creatinylglycine ethylic ether fumarate) on the brain injury was studied in the experimental model of the brain ischemia/reperfusion in rats. The experiments were carried out in 14-20 weeks old male Wistar rats weighing 240-300 g, anesthetized by chloral hydrate (430 mg/kg). Creamide was administered intravenously at the doses of 50, 70, 140, and 280 mg/kg. For comparison phosphocreatine was used at the dose of 255 mg/kg. Creamide and phosphocreatine were administered intravenously (in volume of 1 ml within 5 min) 30 min before cerebral middle artery occlusion. Focal cerebral ischemia for 30 min was produced by endovascular suture occlusion with the subsequent 48 h reperfusion period. Creamide administration resulted in dose-dependent decrease of brain ischemic injury. Creamide administered at the doses of 140 and 280 mg/kg was more effective as compared with phosphocreatine (255 mg/kg). The data obtained open new perspectives for further research and development of new creatine-derived drugs with neuroprotective action.

Topics: Animals; Brain; Brain Ischemia; Disease Models, Animal; Dose-Response Relationship, Drug; Infarction, Middle Cerebral Artery; Injections, Intravenous; Male; Neuroprotective Agents; Phosphocreatine; Rats; Rats, Wistar; Reperfusion Injury; Time Factors

Phosphocreatine-Mg-complex acetate (PCr-Mg-CPLX) is a creatine-derived compound that in previous in vitro research was able to increase neuronal creatine independently of the creatine transporter, thus providing hope to cure the hereditary syndrome of creatine transporter deficiency. In previous research we showed that it reproduces in vitro the known neuroprotective effect of creatine against anoxic damage. In the present paper we investigated if PCr-Mg-CPLX reproduces this neuroprotective effect in vivo, too. We used a mouse model of transient middle cerebral artery occlusion. Mice received PCr-Mg-CPLX or a mixture of the two separate compounds phosphocreatine (PCr) and MgSO(4), or vehicle. The injections were done 60 min and 30 min before ischemia. Forty-eight hours after ischemia neurological damage was evaluated with Clark's behavioural tests, then the infarct volume was measured. PCr-Mg-CPLX reduced the infarct volume by 48%, an effect that was not duplicated by the separate administration of PCr and MgSO(4) and the neurological damage was decreased in a statistically significant way. We conclude that PCr-Mg-CPLX affords in vivo neuroprotection when administered before ischemia. These results are comparable to previous research on creatine administration in experimental stroke. PCr-Mg-CPLX maintains creatine-like neuroprotective effects in vivo as well as in vitro. Our study suggests that PCr-Mg-CPLX might have a therapeutic role in the treatment of hereditary creatine transporter deficiency and of conditions where there is a high risk of impending stroke or cerebral ischemic damage, like high-risk transient ischemic attacks, open heart surgery, and carotid surgery.

Topics: Animals; Brain Infarction; Brain Ischemia; Creatine; Cytoprotection; Disease Models, Animal; Infarction, Middle Cerebral Artery; Magnesium; Male; Membrane Transport Proteins; Mice; Nerve Degeneration; Neuroprotective Agents; Phosphocreatine; Treatment Outcome

Reperfusion injury is believed to contribute to the pathophysiology of ischemic cell death, but the precipitating factors have yet to be completely elucidated. The goal of this study was to examine if reflow-induced secondary energy failure is a component in the events that lead to cell death following increasing periods of middle cerebral artery (MCA) occlusion in Wistar rats. Discrete sections within the MCA distribution were dissected and analyzed for high-energy phosphates and glucose. Regional cerebral blood flow was determined by [14C]-iodoantipyrine technique in representative groups. The levels of ATP + P-creatine were initially depressed at the end of the focal ischemia and the concentrations in the penumbra were unchanged for up to 8 h after 2 h of ischemia which contrasts with response in the ischemic core, striatum, and penumbra where the HEP generally recovered to values near those of control only to decrease with increasing periods of reflow. The possibility of a rebound ischemia in secondary energy failure (SEF) was precluded by regional CBF values and concentrations of glucose that were significantly higher than the threshold for an ischemic effect. The depletion of cellular energy stores following SEF strongly indicates that the evolution of infarct during reflow results from loss of ATP and its synthesis.

Topics: Adenosine Triphosphate; Animals; Brain Ischemia; Cell Death; Cerebrovascular Circulation; Energy Metabolism; Infarction, Middle Cerebral Artery; Male; Mitochondrial Diseases; Nerve Degeneration; Phosphocreatine; Rats; Rats, Wistar; Reperfusion Injury; Telencephalon