Neurodegeneration after transient brain ischemia in aged mice: beneficial effects of bilobalide.

Bilobalide, an active constituent of Ginkgo biloba, has neuroprotective properties in experimental stroke models, but nearly all published studies were carried out in young animals. As ischemic strokes in humans are much more frequent in old age, we investigated bilobalide's effects in aged mice (age 18-22 month) using a model of transient ischemia induced by occlusion of the middle cerebral artery (MCAO) for 60 min. When bilobalide was administered locally into the striatum via microdialysis, a significant reduction of infarct size by almost 70% was observed. Concomitantly, the extensive, twelve-fold increase of extracellular glutamate which was observed in untreated animals was strongly reduced during the infusion of bilobalide. Glucose levels, in contrast, were not affected by bilobalide. In further experiments, bilobalide was given as an intraperitoneal injection (10/mg/kg) 1h before MCAO onset. ATP levels (measured in brain homogenates) were significantly reduced by transient MCAO but pretreatment with bilobalide prevented this loss. In ex vivo experiments with isolated mitochondria from aged mice, we found that the activity of the mitochondrial respiratory chain was only slightly impaired after 60 min of ischemia, and bilobalide showed no benefit in this experiment. However, aged mitochondria proved to be very sensitive to calcium-induced swelling which was significantly increased after ischemia. In this assay, pretreatment with bilobalide lowered the extent of swelling nearly to control levels. In behavioural tests, pretreatment of aged mice with bilobalide significantly improved the outcome in the Rotarod and the Corner test. In conclusion, aged mice show some differences in their response to transient ischemia when compared with young mice. Bilobalide has prominent neuroprotective properties in mice of all ages.

Blood gases and energy metabolites in mouse blood before and after cerebral ischemia: the effects of anesthetics.

The levels of blood gases and energy metabolites strongly influence the outcome of animal experiments, for example in experimental stroke research. While mice have become prominent animal models for cerebral ischemia, little information is available on the effects of anesthetic drugs on blood parameters such as blood gases, glucose and lactate in this species. In this work, we collected arterial and venous blood samples from female CD-1 mice before and after cerebral ischemia induced by middle cerebral artery occlusion (MCAO), and we tested the influence of different anesthetic drugs. We found that all of the injectable anesthetics tested (ketamine/xylazine, chloral hydrate, propofol and pentobarbital) caused a decrease in blood pH and partial pressure of oxygen (pO2) and an increase of partial pressure of carbon dioxide (pCO2), indicating respiratory depression. This was not observed with inhalable anesthetics such as isoflurane, sevoflurane and halothane. Significant and up to two-fold increases of blood glucose concentration were observed under isoflurane, halothane, ketamine/xylazine, chloral hydrate, and propofol anesthesia. Lactate concentration rose significantly by 2-3-fold during inhalation of isoflurane and halothane treatment, but decreased by more than 50% after administration of pentobarbital. Permanent cerebral ischemia induced respiratory acidosis (low pH and pO2, high pCO2) which was most prominent after 24 h. Postsurgical treatment with Ringer-lactate solution (1 mL, intraperitoneal) caused a recovery of blood gases to basal levels after 24 h. Use of isoflurane for surgery caused a minor increase of blood glucose concentrations after one hour, but a strong increase of blood lactate. In contrast, anesthesia with pentobarbital did not affect glucose concentration but strongly reduced blood lactate concentrations one hour after surgery. All values recovered at three hours after MCAO. In conclusion, anesthetic drugs have a strong influence on murine blood parameters, which should be taken into account in experiments in mice.