Protective role of endothelial nitric oxide synthase following pressure-induced insult to the optic nerve.

Although intracranial pressure (ICP) elevation can induce significant structural and functional changes within the central nervous system (CNS), almost complete neuronal recovery is possible if ICP and associated pathogenic factors are restored in the acute phase of the disease process. Nitric oxide synthase (NOS) isoforms have been implicated in the pathogenesis of many CNS diseases and may play an important role in the development of neuronal tolerance in the early stages of pressure elevation. In this paper we use the pig optic nerve, a typical central white matter tract, to study the time-dependent sequence of NOS isoform change following pressure elevation. The timing of NOS isoform change in relationship to structural and functional changes to axons and glial cells is also discussed. This study demonstrates that endothelial cell nitric oxide synthase (ecNOS), an enzyme that plays a protective role in the CNS, is up-regulated in a time-dependent manner after pressure elevation. ecNOS levels increase after axonal and astrocyte injury, suggesting that it might be a compensatory response that is initiated in an effort to preserve CNS function. Inducible NOS (iNOS) and neuronal NOS (nNOS), which are known to have a deleterious effect on the CNS, were not detected in this study. The increase in ecNOS demonstrated in this study is significantly different to the increase in iNOS and nNOS previously demonstrated following traumatic brain injury. Changes in ecNOS levels may therefore be important in the development of neuronal tolerance in the early stages of CNS diseases such as hydrocephalus.

Increased PI3-kinase in presympathetic brain areas of the spontaneously hypertensive rat.

Existing evidence led us to hypothesize that increases in p85alpha, a regulatory subunit of PI3-kinase, in presympathetic brain areas contribute to hypertension. PI3-kinase p85alpha, p110alpha, and p110delta mRNA was 1.5- to 2-fold higher in the paraventricular nucleus (PVN) of spontaneously hypertensive rats (SHR) compared with their controls, Wistar Kyoto rats (WKY). The increase in p85alpha/p110delta was attenuated in SHR treated with captopril, an angiotensin (Ang)-converting enzyme inhibitor, from in utero to 6 months of age. In the rostral ventrolateral medulla (RVLM), p110delta mRNA was approximately 2-fold higher in SHR than in WKY. Moreover, the increases in mRNA were associated with higher PI3-kinase activity in both nuclei. The functional relevance was studied in neuronal cultures because SHR neurons reflect the augmented p85alpha mRNA and PI3-kinase activity. Expression of a p85 dominant-negative mutant decreased norepinephrine (NE) transporter mRNA and [3H]NE uptake by approximately 60% selectively in SHR neurons. In summary, increased p85alpha/p110delta expression in the PVN and RVLM is associated with increased PI3-kinase activity in the SHR. Furthermore, normalized PI3-kinase p85alpha/p110delta expression within the PVN might contribute to the overall effect of captopril, perhaps attributable to a consequent decrease in NE availability.

Effects of magnesium sulfate on Na+,K+ -ATPase and intracranial pressure level after cerebral ischemia.

In the present study, the effects of magnesium sulfate on Na+,K+ -ATPase levels and intracranial pressure (ICP) after cerebral ischemia in rabbits were studied. Thirty New Zealand rabbits were divided into three groups. Group 1 was the control group. In group 2 (untreated group) cerebral ischemia was produced by clamping bilateral common carotid arteries for 60 min but in group 3 magnesium sulfate was administered 100 mg/kg i.v. 10 min after opening the clamps. In group 1, ICP recordings were obtained 5, 60 and 120 min after craniectomy. In groups 2 and 3, ICP recordings were obtained 5 min after craniectomy but before clamping, 60 min after clamping and 60 min after opening the clamps. After taking ICP recordings, brain cortices were resected and Na+,K+ -ATPase activity was determined by subtracting the enzyme activity in the presence of ouabain from the total activity in the absence of ouabain method. There was a significant difference between Na+,K+ -ATPase levels of group 1 and group 2 (P < 0.05). There was no significant difference in Na+,K+ -ATPase levels between group 1 and 3 (P > 0.05), also preischemic ICP values were same in all groups (P > 0.05). Preischemic and postischemic ICP values were significantly different between groups 1 and 2 (P < 0.05), also postischemic (120 min) ICP values were significantly different between group 2 and group 3 (P < 0.05). ICP values correlate well with Na+,K+ -ATPase level. These results demonstrate that cerebral ischemia leads to a decrease of ATPase level in the brain and magnesium sulfate suppresses the decrease of Na+,K+ -ATPase, also magnesium sulfate treatment improves the ICP changes.

Caspase-8 expression and proteolysis in human brain after severe head injury.

Programmed cell death involves a complex and interrelated cascade of cysteine proteases termed caspases that are synthesized as inactive zymogens, which are proteolytically processed to active enzymes. Caspase-8 is an initiator caspase that becomes activated when Fas death receptor-Fas ligand (FasL) coupling on the cell surface leads to coalescence of a "death complex" perpetuating the programmed cell death cascade. In this study, brain tissue samples removed from adult patients during the surgical management of severe intracranial hypertension after traumatic brain injury (TBI; n=17) were compared with postmortem control brain tissue samples (n=6). Caspase-8 mRNA was measured by semiquantitative reverse transcription and polymerase chain reaction, and caspase-8 protein was examined by Western blot and immunocytochemistry. Fas and FasL were also examined using Western blot. Caspase-8 mRNA and protein were increased in TBI patients vs. controls, and caspase-8 protein was predominately expressed in neurons. Proteolysis of caspase-8 to 20-kDa fragments was seen only in TBI patients. Fas was also increased after TBI vs. control and was associated with relative levels of caspase-8, supporting formation of a death complex. These data identify additional steps in the programmed cell death cascade involving Fas death receptors and caspase-8 after TBI in humans.