Junjian Zhang

Cerebral Hypoperfusion and Cognitive Impairment: The Pathogenic Role of Vascular Oxidative Stress

ABSTRACT Alzheimers disease (AD) and vascular dementia (VaD) are the most frequent causes of cognitive impairment in the elderly. In the pathogenesis of cognitive impairment, the association of neurodegenerative and vascular factors indicates a major role of hemodynamic abnormalities including cerebral hypoperfusion. There is also ample evidence that oxidative stress of vascular origin leads to profound alterations in cerebrovascular regulation and is crucial to cerebrovascular dysfunction in a variety of conditions that result in chronic hypoperfusion of the brain. In rodents, experimental chronic cerebral hypoperfusion (CCH) can be initiated by occlusion of the major arterial supply. This way CCH brings about mitochondrial dysfunction and protein synthesis inhibition. These effects may destroy the balance of antioxidases and reactive oxygen species (ROS) and produce oxidative damage. At the same time, oxidative injury to vascular endothelial cell, glia, and neuron impairs vascular function and neurovascular coupling, which may result in a vicious cycle of further reduction of cerebral perfusion. In clinical cases of severe cognitive dysfunction, vascular risk factors are commonly present, while cerebral hypoperfusion is often associated with vascular oxidative damage. Thus we hypothesize that cerebral hypoperfusion is one of the key factors in the development of cognitive impairment, in which vascular oxidative stress plays a major role. The approaches against cerebrovascular dysfunction, combined with antioxidants and others, might make a promising contribution to the treatment of cognitive impairment.

Environmental Enrichment Influences BDNF and NR1 Levels in the Hippocampus and Restores Cognitive Impairment in Chronic Cerebral Hypoperfused Rats

An enriched environment (EE) is beneficial in modifying behaviors, particularly in tasks involving complex cognitive functions. However, the impact of EE on cognitive impairment induced by chronic cerebral hypoperfusion (CCH) has not been studied. We investigated the effects of EE on cognitive impairment caused by CCH and examined whether CCH altered the protein levels of brain-derived neurotrophic factor (BDNF) and N-methyl-D-aspartate (NMDA) receptor subunit 1 (NR1) and subunit 2B (NR2B) in the hippocampus of rats and whether EE exposure attenuated the effects. Rats were divided into four groups that received either permanent bilateral ligation of the common carotid arteries (2-vessel occlusion) surgery or sham surgery followed by either EE housing or standard environment housing for 4 weeks. We examined non-spatial recognition memory in the novel object recognition task, spatial learning, and memory ability in the Morris water maze as well as the protein levels of BDNF, NR1, and NR2B in the hippocampus. CCH impaired both spatial and non-spatial cognitive functions, and EE exposure reversed the spatial cognitive performance and improved non-spatial memory performance. CCH resulted in decreased levels of BDNF and NR1 protein in the hippocampus, and EE exposure restored the decreased expression. Our results demonstrate for the first time that EE exposure restores cognitive impairment induced by CCH and up-regulates the decreased protein levels of BDNF and NR1. Inversely, BDNF and NR1 may contribute to the beneficial effects of EE on CCH in rats.

Downregulation of serum brain specific microRNA is associated with inflammation and infarct volume in acute ischemic stroke

Cerebral ischemic injury activates a robust inflammatory response, exacerbating neurological deficit. Several brain specific microRNA (miRNA) molecules have been reported to mediate functioning of the immune system, referred to as NeurimmiR. We aimed to explore possible associations between serum miRNA levels and stroke severity and their involvement in the regulation of inflammatory responses after stroke. Blood samples were obtained from 31 patients with acute ischemic stroke and 11 healthy controls. We evaluated infarct volume using diffusion weighted imaging and neurological deficit using the National Institutes of Health Stroke Scale. Serum levels of three NeurimmiR, miR-124, miR-9 and miR-219 were detected by real-time polymerase chain reaction and serum levels of metalloproteinase-9 (MMP-9), a proinflammation marker in brain injury, were examined by enzyme-linked immunosorbent assay. We found that serum miR-124 was significantly decreased within 24 hours after stroke onset and serum miR-9 was decreased in patients with larger stroke. There were no significant changes in serum miR-219. Both serum miR-124 and miR-9 levels within 24 hours were negatively correlated with infarct volume and plasma high-sensitivity C-reactive protein levels. All three NeurimmiR negatively correlated with MMP-9 levels. Our preliminary findings indicate that serum miR-124, miR-9 and miR-219 are suppressed in acute ischemic stroke thus facilitating neuroinflammation and brain injury.

Comparison of subjective and objective sleep quality in menopausal and non-menopausal women with insomnia

BACKGROUND Insomnia affects midlife women as they approach and experience menopause at a rate higher than most other stages of life. Insomnia is considered one of the climacteric symptoms of menopause, which can be controlled with hormone replacement therapy (HRT). This study examined the relationship between menopause and sleep in women with insomnia and compared the sleep quality of menopausal women with and without HRT. METHODS A total of 74 women (age range=40-59 years old) with insomnia who were either pre or peri/post menopause were evaluated at Laval Universitys Sleep Disorders Center as part of ongoing clinical trials of insomnia therapies. All participants completed daily sleep diaries for a 2-week period and a series of psychological and insomnia questionnaires, followed by three consecutive nights of polysomnographic evaluation (PSG). A detailed medical history interview was taken by the study physician. RESULTS PSG measures showed that menopausal women had significantly longer total wake time (TWT, 84.2 vs. 63.2 min, Cohens d=0.504) and lower sleep efficiency (SE, 81.8% vs. 86.0%, d=0.487) than the non-menopausal women. Women using HRT did not show significantly better sleep compared to those who did not use HRT. No significant difference was observed between menopausal groups on subjectively assessed sleep parameters. CONCLUSION Menopause may contribute to specific aspects of sleep disturbances in midlife women with insomnia. Use of HRT for menopausal symptoms does not seem to attenuate such disturbances, although further investigation using hormonal level dosing and a larger sample size is warranted.

Regional cerebral blood flow and cerebrovascular reactivity in Alzheimer's disease and vascular dementia assessed by arterial spinlabeling magnetic resonance imaging.

Hemodynamic disturbance in cerebral blood flow (CBF) is common in both Alzheimers disease (AD) and vascular dementia (VaD).The aim of this study is to investigate the different patterns of regional cerebral blood flow (rCBF) change and cerebrovascular reactivity (CVR) in these two types of dementia. Mean flow velocity (MFV) of middle cerebral artery and rCBF were measured by Transcranial Doppler ultrasound (TCD) and arterial spin-labeling (ASL) magnetic resonance, separately. CVR was evaluated by MFV or rCBF change in response to 5% CO2 inhalation. The ASL results showed that, rCBF was significantly lower in both the bilateral frontal and temporal lobes in AD group and lower in left frontal and temporal white matter in patients with VaD. CVR calculated by rCBF was impaired more severely in bilateral frontal cortices in AD. Conversely, TCD tests failed to demonstrate significant difference in MFV and CVR between the two groups. It is concluded that the different patterns detected by ASL in resting rCBF change and cerebrovascular reactivity in response to carbogen inhalation may serve as a potential marker to distinguish AD and VaD.

Mild hypothermia reduces ischemic neuron death via altering the expression of p53 and bcl-2

Abstract Objective: Studies exploring roles of p53 and bcl-2 in neuroprotection by hypothermia in focal cerebral ischemia have not provided consistent results. In the present study, we determined whether p53 and bcl-2 are involved in the hypothermia-induced neuroprotection. Methods: Male Sprague–Dawley rats were divided into four groups: normothermic (37–38°C) ischemia, hypothermic (31–32°C) ischemia, hyperthermic (41–42°C) ischemia and sham-operated group. Global cerebral ischemia was established for 20 minutes using the Pulsinelli four-vessel occlusion model and the brain temperature was maintained at defined levels for 60 minutes following the 20 min ischemia. The mortality in rats was evaluated at 72 hour and 168 hour reperfusion. The expression of p53 and bcl-2 proteins was detected at 24, 48 and 72 hours after reperfusion. At the same intervals, neuron necrosis and apoptosis in brain regions was also detected using hematoxylin and eosin (HE) staining and terminal deoxynucleotldyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL). Results: The mortalities of rats in normothemia, hypothermia and hyperthermia groups was 33·3, 16·7 and 50% at 72 hour reperfusion. At 168 hours of reperfusion, the mortality in the three groups was 58·3, 25 and 100%, respectively. In light microscopy studies, necrotic neurons and apoptotic neurons were found in the hippocampus after global cerebral ischemia. Surviving neurons in hippocampus was increased in mild hypothermic ischemia group (p<0·05) and decreased in hyperthermia ischemia group (p<0·01) at 24, 48 and 72 hour reperfusion. TUNEL-positive neurons in hippocampus decreased in hypothermic ischemia group (p<0·05 or p<0·01) and increased in hyperthermic ischemia group (p<0·01) at 24, 48 and 72 hour reperfusion. The expression of p53 and bcl-2 proteins was found in the neurons of cerebral cortex after global cerebral ischemia. P53 decreased and bcl-2 increased in hypothermia group. Conclusion: Hypothermia reduces ischemic neuronal necrosis and apoptosis by reducing p53 and increasing bcl-2 expression. Hyperthermia accelerated ischemic neuronal injury by increasing p53 and reducing bcl-2 expression.

PI3K/Akt Signal Pathway Involved in the Cognitive Impairment Caused by Chronic Cerebral Hypoperfusion in Rats

Chronic cerebral hypoperfusion (CCH) is a common pathophysiological state that usually occurs in conditions such as vascular dementia and Alzheimers disease, both of which are characterized by cognitive impairment. In previous studies we found that learning capacity and memory were gradually impaired with CCH, which altered the expression of synaptophysin, microtubule associated protein-2, growth associated protein-43, brain-derived neurotrophic factor, nerve growth factor, N-methyl-D-aspartate receptor subunit 1, cAMP response element-binding protein and tau hyperphosphorylation in the hippocampus. However, the molecular basis of cognitive impairment in CCH remains obscure. Here we explore the hypothesis that the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signal pathway is involved in this type of cognitive impairment. In order to determine if the expression of PI3K, Akt and phosphorylated Akt (p-Akt) proteins are altered at different stages of CCH with differing levels of cognitive impairment. we performed permanent, bilateral occlusion of the common carotid arteries (2-VO) to induce CCH. Adult male SD rats were randomly divided into sham-operated group, 2-VO 1 week group, 2-VO 4 weeks group and 2-VO 8 weeks group. Behavior tests were utilized to assess cognitive abilities, while western blots were utilized to evaluate protein expression. Rats in the 2-VO groups spent less time exploring novel objects than those in the sham-operated group, and the discrimination ratio of the 2-VO 8 weeks group and the sham-operated group were higher than chance (0.50). Escape latencies in the Morris water maze task in the 2-VO 1 week group were longer than those in the sham-operated group on day 4 and day 5, while escape latencies in the 2-VO 4 weeks group were longer than those in the sham-operated group from day 3 to day 5. Escape latencies in 2-VO 8 weeks group were longer than those in the sham-operated group from day 2 to day 5. NE (northeast) square swimming times in the 2-VO 1 week group, 2-VO 4 weeks group and 2-VO 8 weeks group were shorter than that in the sham-operated group. Western blotting showed that the PI3K expression in the 2-VO 1 week group was lower than that in sham-operated group, while p-Akt expression in the 2-VO 8 weeks group was higher than that in the sham-operated group. There was a linear relationship between the PI3K expression and the discrimination ratio, as well as a linear relationship between the PI3K and NE square swimming time. Thus, we propose that the PI3K/Akt signal pathway is an important cell pathway that is associated with the cognitive impairment following CCH.

An enriched environment reverses the synaptic plasticity deficit induced by chronic cerebral hypoperfusion

Chronic cerebral hypoperfusion (CCH) leads to a long-term, inadequate blood supply in the brain, which eventually causes cognitive impairment. An enriched environment (EE) improves learning and memory by improving synaptic plasticity. The impact of an EE on cognitive impairment induced by CCH is not, however, well known. To investigate this possible effect, we permanently occluded the bilateral common carotid arteries (2-vessel occlusion) in rats to induce CCH and studied EE effects on cognitive impairment and synaptic plasticity following CCH. We found that EE treatment reversed spatial memory deficits induced by CCH. An EE also reversed the deficit in long-term potentiation following CCH, but the input-output curves and paired-pulse facilitation were not affected. CCH led to reduced expression of phosphorylated CREB in the rats, but EE reversed this reduction. In addition, CCH reduced the expression of synaptophysin and microtubule-associated protein 2, whereas EE reversed this reduced expression. Thus, EE reversed CCH-induced spatial cognitive impairment without affecting basal synaptic transmission or the release probability of presynaptic neurotransmitters. The EE effect probably resulted from the regulation of postsynaptic potentiation.

Therapeutic effect of post-ischemic hypothermia duration on cerebral ischemic injury

Abstract Objective: To study the efficacy of post-ischemic mild brain hypothermia lasting for different time intervals on cerebral ischemic reperfusion injury. Method: Male Sprague–Dawley rats were divided into a sham-operated group, normothermia (37–38° C) ischemia group and mild hypothermia (31–32° C) group. The last group was subdivided into four groups: 30 minute hypothermia plus 210 minute normothermia, 60 minute hypothermia plus 180 minute nomothermia,120 minute hypothermia plus 120 minute normothermia, and 240 minute hypothermia (n=8). Global cerebral ischemia was established using the Pulsinelli four-vessel occlusion model. Brain tissue was collected following a 20 minute cerebral ischemia and 240 minute reperfusion, and was used to measure the levels of glutamate (Glu), aspartate (Asp), glycine (Gly), gamma-aminobutyric acid (GABA), dopamine (DA), norepinephrine (NE), serotonin(5-HT) and hydroxyindoleacetic acid (5-HIAA), nitrite (NO2), endothelin-1 (ET1), tumor necrosis factor alpha(TNFα) and interleukin-1beta (IL-1β). Serum was collected to measure the levels of lactate dehydrogenase (LDH), aspartate aminotransferase (AST), creatine kinase (CK) and its brain band isoenzyme (CK-BB). Results: Hypothermia lasting for 60–240 minutes delayed the decrease in these amino acids, postponed the decrease in DA, NE and 5-HT and increase in hydroxyindoleacetic acid (5-HIAA), and decreased the levels of IL-1β, TNFα, ET1 and NO2 in brain tissue. Hypothermia also decreased the levels of LDH, AST, CK and CK-BB in serum as compared to normothermia group (p<0.05 or p<0.01). Hypothermia lasting for 30 minutes delayed the decreases in these amino acids and 5-HT and increase in 5-HIAA in brain tissue (p<0.05), but failed to influence the levels of IL-1β, TNFα, ET1 and NO2 in brain tissue and the amounts of LDH, AST, CK and CK-BB in serum as compared to normothermia ischemia group (p>0.05). Conclusions: Post-ischemic mild brain hypothermia can significantly suppress the excessive release of amino acids, monoamine neurotransmitters and inflammation response in ischemic tissue. It can also stabilize the function of the cell membrane, which is associated with the mechanism of cerebral protection by mild hypothermia. These results suggest that mild hypothermia should be applied immediately after ischemia and last for more than 60 minutes in order to obtain neuroprotective effects.

Enriched Environment Prevents Cognitive Impairment and Tau hyperphosphorylation after Chronic Cerebral Hypoperfusion

Chronic cerebral hypoperfusion (CCH) has been gradually prevalent in the patients over middle age, especially the old over 60 years. It has been proved that CCH is highly related with cognitive impairment. CCH emerges not only in vascular dementia (VaD), but also in Alzheimers disease (AD), which regarded as a critical causative for cognitive impairment in these diseases. Nevertheless, the mechanisms underlying cognitive deficit remain elusive. Moreover, there are no dramatically effective preventions. In the present study, by employing a recognized CCH rat model, we found that CCH induced spatial learning/memory deficits with simultaneously increasing tau hyperphosphorylation at multiple Alzheimer-related phosphorylation sites with activation of glycogen synthase kinase-3β (GSK-3β), Cyclin-dependent kinase (Cdk5), Calcium/calmodulin-dependent protein kinase II (CaMKII), and protein kinase B (Akt), and inhibition of protein phosphatase (PP) 2A (PP-2A). Interestingly, enriched environment (EE) treatment, an effect environment stimuli filled with various novel objects, could prevent rats from the EE-induced memory deficits and alterations of tau hyperphosphorylation. Our data suggested that EE might be potentially used for attenuating the detrimental cognition induced by CCH through regulating tau hyperphosphorylation.