oxypurinol and Brain-Ischemia

Oxygen free radicals, generated by cerebral ischemia, have been widely implicated in the damage of vascular endothelium. Endothelial cells have been proposed as a significant source of oxygen free radicals. In the present study, we developed an anoxia-reoxygenation (AX/RO) model using pure cultures of cerebral endothelial cells (CECs) isolated from piglet cortex to measure CEC oxygen free radical production and determine its role in AX/RO-induced CEC injury. CEC injury, as measured by lactate dehydrogenase efflux into the culture medium, increased progressively with the duration of anoxic exposure, becoming significant after 10 h. Reoxygenation significantly increased CEC anoxic injury in a time-dependent manner. A 55% increase in oxygen free radical production, determined by fluorescence detection of dihydroethidium oxidation, was measured at the end of 4-h reoxygenation in CECs subjected to AX/RO conditions that killed 40% of the cells. Blockade of oxygen free radical production with superoxide dismutase (SOD; 250 and 1000 U/ml) or oxypurinol (50 and 200 microM), a potent xanthine oxidase inhibitor, reduced this injury by 32-36% and 30-39%, respectively. Results from our in vitro model indicate that CECs produce significant amounts of oxygen free radicals following ischemia, primarily from the xanthine oxidase pathway. These radicals ultimately have a cytotoxic effect on the very cells that produced them. Thus, reductions in oxygen free radical-mediated vascular injury may contribute to improvements in neurophysiologic outcome following treatment with oxygen free radical inhibitors and scavengers.

Topics: Animals; Brain Ischemia; Cells, Cultured; Cerebrovascular Circulation; Endothelium, Vascular; Enzyme Inhibitors; Microcirculation; Oxypurinol; Reperfusion Injury; Superoxide Dismutase; Superoxides; Swine; Xanthine Oxidase

The superoxide (O2.-) scavenging activity and neuroprotective effects of oxypurinol, a xanthine oxidase inhibitor, were compared with those of alpha-phenyl-N-tert-butyl nitrone (PBN). The rate constant for the reaction of oxypurinol with O2.- at pH 7.4 was 1.71 x 10(3) M(-1) s(-1) which was more than 100-fold that of PBN (1.65 x 10 M(-1) s(-1)). Oxypurinol inhibited the release of O2.- from stimulated neutrophils better than did PBN. However, oxypurinol did not attenuate the ischemic neuronal damage in gerbils, while PBN did. These results indicate that neither xanthine oxidase inhibiting activity nor O2.- scavenging activity correlates to the therapeutic efficacy of neuroprotective agents in ischemic-reperfusion injury.

Topics: Animals; Brain Ischemia; Cyclic N-Oxides; Electron Spin Resonance Spectroscopy; Enzyme Inhibitors; Gerbillinae; Kinetics; Male; Neurons; Neutrophils; Oxypurinol; Spin Labels; Superoxides; Tetradecanoylphorbol Acetate; Xanthine Oxidase

Topics: Animals; Brain Ischemia; Cerebral Cortex; Electron Spin Resonance Spectroscopy; Hydroxides; Male; Oxypurinol; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Reperfusion; Time Factors

When measured within 2 days of a unilateral occlusion of the middle cerebral artery (MCA) combined with tandem occlusion of the ipsilateral common carotid artery in rats, contralateral neurological deficits were detectable, with brain swelling and a consistent degree of neocortical infarction in the ipsilateral hemisphere. Oxypurinol (40 mg/kg i.p. administered 0.5 h prior to, and 24 h after, the onset of focal ischemia) significantly reduced the development of the ischemic infarct (P less than 0.001); attenuated tissue swelling (P less than 0.01) and ameliorated the neurological deficits (P less than 0.05). These findings suggest that this compound may be useful for the prevention or treatment of ischemic brain injuries, such as those occurring during stroke.

Topics: Animals; Brain Edema; Brain Ischemia; Cerebral Infarction; Male; Multivariate Analysis; Nervous System; Nervous System Diseases; Oxypurinol; Physical Stimulation; Rats; Rats, Inbred F344