Current status and future prospects of pathophysiology-based neuroprotective drugs for the treatment of vascular dementia.

Vascular dementia (VaD) is a progressive neurocognitive clinical syndrome that is caused by a decrease in cerebral blood flow and damage to the neurovascular unit. Given increasing life expectancy, VaD is emerging as one of the leading health problems in society. Despite the high global prevalence of cognitive impairment associated with VaD, diagnosis and treatment still remain limited because of the complexity of mechanisms of neuronal loss. Therefore, advances in our understanding of the pathophysiological mechanisms involved is crucial for the development of new therapeutic strategies. In this review, we highlight the pathophysiology, current pharmacology-based primary and secondary prevention strategies and emerging treatment options for VaD.

The role of neutrophils in mediating stroke injury in the diabetic db/db mouse brain following hypoxia-ischemia.

Diabetic mice exhibit increased mortality and morbidity following stroke. Recent studies from our laboratory have indicated that increased morbidity in diabetic db/db mice relative to their non-diabetic db/+ littermates is associated with increased levels of MMP-9 protease activity, increased blood-brain barrier (BBB) permeability, and greater neutrophil infiltration following hypoxic/ischemic (H/I) insult. Neutrophils are a major source of proteases and reactive oxygen species and studies have reported neutrophil depletion/inhibition is protective in certain models of experimental stroke. The objective of the current study is to determine the role of neutrophils in the increased morbidity seen in db/db mice following acute ischemic stroke. In this study, we found a significant increase in circulating neutrophils in the db/db mice at 4 h post H/I, which bound to endothelial cells in the ipsilateral hemisphere and infiltrated into brain tissue by 24 h of recovery. Depletion of circulating neutrophils resulted in reduced neutrophil concentrations in blood and in the ipsilateral hemispheres of the brain of both db/+ and db/db mice and decreased the levels of MMP-9 within the infarcted area. This resulted in smaller infarct size in the db/db mice compared to non-treated controls but did not affect stroke outcome in db/+ mice. While there was a significant correlation between neutrophil number and the levels of MMP-9 in the ipsilateral hemisphere of control and diabetic mice, surprisingly, neutrophil depletion had no effect on BBB permeability in either group. Thus, the current study suggests that neutrophil depletion reduces MMP-9 protease levels and improves stroke outcome in db/db mice but not in their db/+ counterparts.

Clinical outcomes of moderate to severe acute ischemic stroke in a telemedicine network.

Acute ischemic strokes (AIS) with severe neurologic deficits are associated with poor short- and long-term prognosis. Thrombectomy alone or in combination with thrombolysis is used for reperfusion in patients with moderate-to-severe AIS. However, the best therapeutic approach within the setting of telemedicine networks needs to be elucidated further. The objective of this study was to analyze clinical and imaging based outcomes of moderate to severe stroke following treatment with thrombolysis, thrombectomy or a combination of both in a telemedicine network. Data of this retrospective study was abstracted from the institutional telestroke database. Patients with a National Institute of Health Stroke Scale score (NIH-SS ≥ 10) were included into the study. Primary outcome measure was the difference in NIH-SS at admission compared to discharge from the hospital. Secondary outcome measure was the discharge disposition defined as favorable (discharge to home or rehabilitation) versus unfavorable disposition (discharge to hospice/death). Furthermore, outcome was analyzed based on reperfusion status following thrombectomy using the Thrombolysis in Cerebral Infarction (TICI) scale. The NIH-SS improved in all three groups, independent of treatment subtype, with a trend towards best outcomes following thrombolysis and combined treatment therapy compared to thrombectomy alone. In addition, reperfusion rates were higher in the combination group compared to the thrombectomy only group. The number of favorable discharges was similar in all three groups. The present study stresses the benefits of tele-stroke networks in allowing to early identify and treat even patients with severe strokes and benefit from different treatment modalities.

Transient focal ischemia induces motor deficit but does not impair the cognitive function in middle cerebral artery occlusion model of stroke in rats.

The effect on cognition was evaluated in the transient ischemia model of stroke in rats. Male Wistar rats were subjected to 2 h of transient ischemia by occluding the middle cerebral artery using the 4-0 intraluminal nylon thread. Neurological tests performed after 24 h, 7 and 15 days of middle cerebral artery (MCA) occlusion showed motor impairment as evidenced by decrease in the rota rod performance test. This was also confirmed histologically. However, when the learning and memory tests were performed, no change was observed in the learning and behavior as evidenced by insignificant difference in the retention latency in the passive avoidance test (334+/-31 s) and the transfer latency in the elevated plus maze (11+/-4.5 s) as compared to the sham-operated rats 369+/-34 and 8+/-1.7 s, respectively. The results of the present study demonstrates that transient occlusion of middle cerebral artery does not impair the learning and memory behavior of the rats.

Protective effect of resveratrol against oxidative stress in middle cerebral artery occlusion model of stroke in rats.

Free radicals have been implicated in neuronal injury during ischemia reperfusion in stroke. Trans resveratrol, a potent antioxidant, polyphenolic compound found in grapes and wines has recently been shown to have neuroprotective activity against oxidative stress in in vitro studies. In the present study the effect of chronic treatment of trans resveratrol was evaluated in focal ischemia induced by middle cerebral artery [MCA] occlusion in rats. Male Wistar rats were pretreated with trans resveratrol 20 mg/kg i.p. for 21 days and were subjected to focal ischemia by occlusion of MCA using intraluminal thread. After two hours of MCA occlusion reperfusion was allowed by retracting the thread. Animals were assessed for motor performance after 24 hours and subsequently rats were sacrificed for estimation of markers of oxidative stress [malondialdehyde [MDA] and reduced glutathione] and for evaluation of volume of infarction. Control group received vehicle and similar protocol was followed. Significant motor impairment, with elevated levels of MDA and reduced glutathione was observed in the vehicle treated MCA occluded rats. Treatment with trans resveratrol prevented motor impairment, rise in levels of MDA and reduced glutathione and also significantly decreased the volume of infarct as compared to control. The study provides first evidence of effectiveness of trans resveratrol in focal ischemia most probably by virtue of its antioxidant property.

Protective effect of exogenous administration of alpha-tocopherol in middle cerebral artery occlusion model of cerebral ischemia in rats.

Oxidative stress due to increased free radical generation and depletion of endogenous antioxidants have been implicated in ischemia-reperfusion-induced neuronal injury. The present study was carried out to study the effect of acute administration of alpha-tocopherol in the middle cerebral artery (MCA) occlusion model of stroke in rats. Rats were anesthetized using chloral hydrate (400 mg/kg i.p.) and subjected to 2 h of transient MCA occlusion. alpha-Tocopherol was administered at the dose of 125 and 250 mg/kg orally 1 h prior to the occlusion of MCA. Motor performance test (grip test, foot fault test, rotarod performance test, spontaneous locomotor activity), markers of oxidative stress and 2,3,5-triphenyl tetrazolium chloride staining were carried out 24 h after MCA occlusion. A vehicle-treated group was run parallel. It was observed that alpha-tocopherol at the dose of 125 mg/kg neither improved neurologic deficit, nor decreased the raised level of oxidative stress markers in comparison with the MCA-occluded rats. However, higher dose of alpha-tocopherol (250 mg/kg p.o.) afforded significant protection as evident by increase in motor performance tests and a decrease in the volume of infarction. The raised levels of malondialdehyde after MCA occlusion were also significantly attenuated. The results demonstrate that exogenous administration of alpha-tocopherol is able to reduce the neuronal damage caused during ischemia-reperfusion, which can be attributed to its antioxidant activity.

Protective effect of adenosine against neuronal injury induced by middle cerebral artery occlusion in rats as evidenced by diffusion-weighted imaging.

In the present study, adenosine, an inhibitory neuromodulator, was studied in male Wistar rats subjected to 2 h of transient middle cerebral artery (MCA) occlusion. Adenosine (500 mg/kg ip) was administered twice-once at the time of MCA occlusion and again at the time of reperfusion-and evaluated for its protective effect by using diffusion-weighted imaging (DWI) (30 min after reperfusion). After the DWI experiments, one group of animals was euthanized 2 h after reperfusion for the estimation of oxidative stress markers, while in another group, neurological deficit was assessed 24 h after MCA occlusion. In the adenosine-treated group, percent hemispheric lesion area (%HLA) in DWI was significantly attenuated (11.7+/-5.2) as compared to vehicle-treated group (21.4+/-4.7). The level of malondialdehyde (MDA) (301.8+/-22 nmol/g wet tissue) in the adenosine-treated group was significantly decreased as compared to that in the vehicle-treated MCA-occluded rats (420+/-20 nmol/g wet tissue). An insignificant change was observed in the levels of glutathione in both the vehicle-treated MCA-occluded and the adenosine-treated groups. The neurological deficit was significantly improved in the adenosine-treated group (1.8+/-0.06) as compared to the vehicle-treated (2.9+/-0.38) group. This is the first study to demonstrate the effectiveness of adenosine using DWI in the MCA-occluded rats.

Functional changes in genetically dysmyelinated spinal cord axons of shiverer mice: role of juxtaparanodal Kv1 family K+ channels.

Axonal dysfunction after spinal cord injury (SCI) and other types of neurotrauma is associated with demyelination and exposure of juxtaparanodal K+ channels. In this study, sucrose gap electrophysiology using selective and nonselective K+ channel blockers, confocal immunohistochemistry, and Western blotting were used to study the role of Kv1.1 and Kv1.2 K+ channel subunits in dysmyelination-induced spinal cord axonal dysfunction in shiverer mice, which lack the gene encoding myelin basic protein (MBP) and exhibit incomplete myelin sheath formation on CNS axons. The shiverer spinal cord axons exhibited smaller amplitude of compound action potentials (CAPs), reduced conduction velocity, reduced excitability, and greater degree of high-frequency conduction failure. The "fast" K+ channel blocker 4-aminopyridine, the toxin DTX-I, which targets the Kv1.1 and Kv1.2, but not DTX- K, which has higher selectivity for Kv1.1, increased the amplitude and area of CAPs of shiverer mice spinal cord axons but had insignificant effects in wild-type mice. Confocal immunohistochemistry showed that, unlike wild-type mice, which have a precise juxtaparanodal localization of the Kv1.l and Kv1.2 K+ channel subunits, shiverer mouse axons displayed a dispersed distribution of these subunits along the internodes. In contrast, the Kv1.l and Kv1.2 subunits, Na+ channels remained highly localized to the nodal regions. Western blotting showed an increased expression of Kv 1.1 and 1.2 in the shiverer mouse spinal cord. These results provide evidence that the neurological deficits associated with myelin deficiency reflect the altered distribution and expression of the K+ channel subunits Kv1.l and Kv1.2 along the internodes of spinal cord axons associated with the biophysical consequences caused by alterations in the myelin sheaths.

Structural and functional alterations of spinal cord axons in adult Long Evans Shaker (LES) dysmyelinated rats.

Abnormal formation or loss of myelin is a distinguishing feature of many neurological disorders and contributes to the pathobiology of neurotrauma. In this study we characterize the functional and molecular changes in CNS white matter in Long Evans Shaker (LES) rats. These rats have a spontaneous mutation of the gene encoding myelin basic protein which results in severe dysmyelination of the central nervous system (CNS), providing a unique model for demyelinating/dysmyelinating disorders. To date, the functional and molecular changes in CNS white matter in this model are not well understood. We have used in vivo somatosensory evoked potential (SSEP), in vitro compound action potential (CAP) recording in isolated dorsal columns, confocal immunohistochemistry, Western blotting and real-time PCR to examine the electrophysiological, molecular and cellular changes in spinal cord white matter in LES rats. We observed that dysmyelination is associated with dispersed labeling of Kv1.1 and Kv1.2 K+ channel subunits, as well as Caspr, a protein normally confined to paranodes, along the LES rat spinal cord axons. Abnormal electrophysiological properties including attenuation of CAP amplitude and conduction velocity, high frequency conduction failure and enhanced sensitivity to K+ channel blockers 4-aminopyridine and dendrotoxin-I were observed in spinal cord axons from LES rats. Our results in LES rats clarify some of the key molecular, cellular and functional consequences of dysmyelination and myelin-axon interactions. Further understanding of these issues in this model could provide critical insights for neurological disorders characterized by demyelination.

Enhanced protection by melatonin and meloxicam combination in a middle cerebral artery occlusion model of acute ischemic stroke in rat.

Mixed efficacy of neuroprotective drugs in clinical trials has led to the emergence of the approach of combination therapy in stroke. The present study was carried out to investigate the effect of the combination of melatonin (potent antioxidant) and meloxicam (preferential inhibitor of cyclooxygenase-2 enzyme) against a middle cerebral artery occlusion model of stroke in rats. Male Wistar rats in the weight range of 250-300 g were used. Rats were anesthetized using chloral hydrate (400 mg/kg i.p) and subjected to 2 h of transient middle cerebral artery occlusion. Melatonin was administered at a dose of 20 mg/kg i.p. four times: at the time of middle cerebral artery occlusion, 1.5 h after middle cerebral artery occlusion, at the time of reperfusion, and 1 h after reperfusion. Meloxicam (2.5 mg/kg) was administered 4 h after middle cerebral artery occlusion. Motor performance tests (grip test, foot fault test, rotarod performance test, spontaneous locomotor activity), markers of oxidative stress, and triphenyltetrazolium chloride staining were carried out 24 h after middle cerebral artery occlusion. A vehicle-treated group was run in parallel. It was observed that melatonin treatment improved the motor performance and significantly attenuated the levels of malondialdehyde (MDA) as compared with the middle cerebral artery occluded group. Meloxicam treatment at the dose used neither showed significant improvement on the motor performance nor decreased the levels of MDA significantly as compared with the middle cerebral artery occluded group. However, when the combination of the two drugs was used, better protection was observed as was evident by the significant decrease in the percent foot fault errors, the increase in the time spent on the rotarod, and the increase in the six-point neurological score and grip test score. There was also a significant decrease in the levels of MDA in the combination group. The results of the present study demonstrate that enhanced protection is observed with the use of a combination of melatonin plus meloxicam in the middle cerebral artery occlusion model of acute ischemic stroke in rats.