Sirtuin 1 activation enhances the PGC-1α/mitochondrial antioxidant system pathway in status epilepticus.

Sirtuin 1 (SIRT1) regulates numerous neuronal processes, including metabolism, antioxidation and aging, through activation of peroxisome proliferator-activated receptor coactivator 1-α (PGC-1α), an upstream regulator of mitochondrial biogenesis and function. However, the role of SIRT1 in the oxidative stress induced by seizures has yet to be elucidated. The present study aimed to investigate whether SIRT1 was involved in the activation of the PGC-1α/mitochondrial antioxidant system following status epilepticus (SE) in rats. The data demonstrated that SIRT1 expression and activity were enhanced in the rat hippocampus following SE. SIRT1 inhibition effectively blocked the SE-associated increase in PGC-1α and mitochondrial antioxidant enzymes, including superoxide dismutase 2 (SOD2) and uncoupling protein 2 (UCP2). Additionally, it was also demonstrated that the activation of SIRT1 enhanced mitochondrial electron transport chain complex I activity and increased ATP content. In conclusion, the present results suggest that SIRT1 activation may alleviate mitochondrial oxidative stress induced by seizures partially via PGC-1α signaling.

Abnormal expression of synaptophysin, SNAP-25, and synaptotagmin 1 in the hippocampus of kainic acid-exposed rats with behavioral deficits.

Temporal lobe epilepsy is characterized by spontaneous recurrent seizures (SRS) and associated with behavioral problems. However, the molecular mechanisms underlying these problems are not yet clear. In this study, kainic acid (KA) was systemically administered to immature male Wistar rats to induce SRS. The behavior of the immature rats was evaluated with a water maze, elevated-plus mazes, and open field tests. The expression patterns of synaptophysin, SNAP-25, and synaptotagmin 1 (Syt 1) were examined by reverse-transcriptase polymerase chain reaction (RT-PCR) and Western blot analysis. KA-treated rats with SRS demonstrated learning and memory deficits, reduced anxiety, and increased locomotor activity, compared with placebo-treated rats and KA-treated rats without SRS. No neuronal cell loss was observed in the hippocampus 6 weeks after exposure to KA. However, RT-PCR and Western blot analyses revealed decreased synaptophysin, SNAP-25, and Syt 1 expression in KA-treated rats with SRS. Synaptophysin, SNAP-25, and Syt1 expression levels were found to be positively correlated with learning and memory but negatively correlated with anxiety and locomotor activity. These data suggested that SRS may induce changes in synaptophysin, SNAP-25, and Syt1 expression and may be functionally related to SRS-induced behavioral deficits.

Features of a Chinese family with cerebral cavernous malformation induced by a novel CCM1 gene mutation.

BACKGROUND: Familial cerebral cavernous malformations (CCMs), characterized by hemorrhagic stroke, recurrent headache and epilepsy, are congenital vascular anomalies of the central nervous system. Familial CCMs is an autosomal dominant inherited disorder and three CCM genes have been identified. We report a Chinese family with CCMs and intend to explore clinical, pathological, magnetic resonance imaging (MRI) features and pathogenic gene mutation of this family. METHODS: Totally 25 family members underwent brain MRI examination and clinical check. Two patients with surgical indications had surgical treatment and the specimens were subjected to histopathological and microstructural examination. In addition, polymerase chain reaction (PCR) and direct sequencing were performed with genomic DNA extracted from 25 family members' blood samples for mutation detection. RESULTS: Brain MRI identified abnormal results in seven family members. All of them had multiple intracranial lesions and four cases had skin cavernous hemangioma. T2-weighted sequence showed that the lesions were typically characterized by an area of mixed signal intensity. Gradient-echo (GRE) sequence was more sensitive to find micro-cavernous hemangiomas. There was a wide range in the clinical manifestations as well as the age of onset in the family. The youngest patient was an 8-year-old boy with least intracranial lesions. Histopathological and microstructural examination showed that CCMs were typically discrete multi-sublobes of berry-like lesions, with hemorrhage in various stages of illness evolution. They were formed by abnormally enlarged sinusoids and the thin basement membranes. A novel T deletion mutation in exon 14 of CCM1 gene was identified by mutation detection in the seven patients. But unaffected members and healthy controls did not carry this mutation. CONCLUSIONS: The clinical manifestations were heterogenic within this family. We identified a novel mutation (c.1396delT) was the disease-causing mutation for this family and extended the mutational spectrum of CCMs.

The value of T(2) (*)-weighted gradient echo imaging for detection of familial cerebral cavernous malformation: A study of two families.

The aim of this study was to investigate the value of T(2) (*)-weighted gradient echo imaging (GRE T(2) (*)-WI) for the detection of familial cerebral cavernous malformation (FCCM). Twenty-six members of 2 families with FCCM were examined using computed tomography (CT), conventional magnetic resonance imaging (MRI) and GRE T(2) (*)-WI sequences. We identified 12 cases of FCCM using GRE T(2) (*)-WI sequences. These 12 patients had multiple lesions (mean 23). The lesions were most commonly located in the ganglia. Other areas included the cortex-subcortex, thalamus, cerebellum and brainstem. These lesions appeared as a reticulated core of mixed signal intensity with a surrounding rim of decreased signal intensity representing hemosiderin from previous hemorrhages. The mean numbers of lesions and cases of FCCM identified by various conventional MRI sequences were 5-17 and 3-9, respectively. Conventional MRI examination involved T(1)-weighted imaging (T(1)WI), T(2)-weighted imaging (T(2)WI), T(2)-fluid-attenuated inversion recovery (T(2)Flair), diffusion-weighted imaging (DWI) and spin-echo imaging (SE) sequences, in that order. The numbers of lesions identified by MRI were fewer than those identified by GRE T(2) (*)-WI. CT only identified 3 cases with large lesions combined with hemorrhage and calcification. These findings suggest that GRE T(2) (*)-WI is the first choice when diagnosing FCCM compared with CT and conventional MRI.

Resveratrol pre-treatment reduces early inflammatory responses induced by status epilepticus via mTOR signaling.

Resveratrol is indicated to be involved in neuroprotection and anti-inflammation in epileptic rats. The molecular mechanism is still not fully understood. In this study, we investigated the role of resveratrol in nuclear factor-kappa B associated inflammatory responses induced by status epilepticus. Our data showed that seizures activated mammalian target of rapamycin (mTOR), increased the activity of nuclear factor-kappa B and promoted the expressions of inflammatory molecules including inducible nitric oxide synthase, cyclooxygenase-2 and interleukin-1ß. Futhermore, resveratrol significantly inhibited the activation of nuclear factor-kappa B and the production of proinflammatory molecules via mTOR pathway. Additionally, we also proved that the inhibition of mTOR signal by resveratrol was mostly attributed to AMP-activated kinase (AMPK) activation. Altogether, our results suggest that resveratrol suppresses inflammatory responses induced by seizures partially via AMPK/mTOR pathway.

"Effects of baicalin on protease-activated receptor-1 expression and brain injury in a rat model of intracerebral hemorrhage".

"Baicalin, a major flavonoid compound isolated from the dry roots of Scutellaria baicalensis Georgi, has been shown to be neuroprotective after ischemic brain injury. However, little is known about its effects on brain injury following intracerebral hemorrhage (ICH). In this study, we evaluated the effects of baicalin on ICH-induced brain injury in an ICH rat model. Male Wistar rats were injected intracerebrally with 0.5 U collagenaseVII to induce ICH, while control rats were injected with an equal volume of saline. After ICH induction, the rats were randomly divided into four groups and treated with baicalin at different doses (0, 25, 50 or 100 mg/kg) through peritoneal injection. The control rats were injected with an equal volume of vehicle. Brain tissues around the hemorrhage areas were collected on day 1, 3, 5 and 10 after treatment. Brain water content was analyzed by desiccation method; mRNA and protein levels of brain protease-activated receptor-1 (PAR-1) were determined by RT-PCR and Western blot, respectively; cell apoptosis was evaluated by terminal transferase dUTP nick end labeling staining. The results showed that baicalin effectively attenuated brain edema and inhibited cell apoptosis following ICH in a dose- and time-dependent manner, with concomitant suppression of PAR-1 expression at both the mRNA and protein levels. These findings indicate that baicalin has protective effects on ICH-induced brain injury. The effects of baicalin may involve a mechanism of inhibition of PAR-1 expression."

Alternating hemiplegia of childhood in chinese following long-term treatment with flunarizine or topiramate.

OBJECTIVE: Alternating hemiplegia of childhood (AHC) is a rare and intractable disorder. The etiology and standard therapy of AHC remain unknown. The long-term effects of flunarizine or topiramate on patients with AHC are still not clear. METHODS: Fifteen patients were investigated in this study. Their neurological disturbance and mental retardation after drug therapy were evaluated. RESULTS: Nine patients treated with flunarizine therapy and three children with topimarate treatment presented with shorter duration or less frequency of the hemiplegic attacks. These drug responsive patients also showed improvements on neurological disturbance including eye movement disorder, choreoathetotic movements, dystonia, and ataxia. However, seizure episodes and cognitive impairments were not alleviated in AHC with long-term drug therapy. CONCLUSIONS: The findings from the present study support flunarizine or topitamate as the rational treatment for AHC.

Increased expressions of TLR2 and TLR4 on peripheral blood mononuclear cells from patients with Alzheimer's disease.

BACKGROUND: Toll-like receptors 2 (TLR2) and TLR4 are involved in the microglia-mediated inflammatory response, Aß plaque formation and Aß clearance in Alzheimer's disease (AD). Our previous studies have shown that variants in the TLR2 and TLR4 genes are associated with the risk of AD. Therefore, we hypothesize that there may be significant changes in TLR2 and TLR4 expressions on peripheral blood mononuclear cells (PBMCs) from patients with AD when compared to healthy control subjects. METHODS: Sixty patients with late-onset AD (LOAD) and 60 healthy controls matched for sex and age were recruited. Flow cytometry (FCM) was used to detect expressions of TLR2 and TLR4 proteins and real-time quantitative RT-PCR was performed to determine TLR2 and TLR4 mRNAs. RESULTS: Compared with controls, expressions of TLR2 and TLR4 mRNAs were up-regulated in LOAD patients (TLR2/beta-actin mRNA: 0.390±0.204 versus 0.281±0.167, P<0.01; TLR4/beta-actin mRNA: 0.503±0.195 versus 0.322±0.183, P<0.01). The proteins levels were higher in LOAD patients than in controls (TLR2: 97.12±1.67% versus 41.07±18.44%, P<0.01, TLR4: 66.56±23.74% versus 14.83±4.31, P<0.01). In both cases, either AD or control group, TLR2 and TLR4 mRNAs expressions were positively correlated with the levels of proteins (TLR2: r=0.980 and 0.976,P<0.01; TLR4: r=0.938 and 0.970, P<0.01), respectively. There were significant negative correlations between TLR levels and MMSE score (TLR2: r=-0.32; P=0.01; TLR4: r=-0.29; P=0.02). In addition, CC genotype can increase the expression of TLR4 in AD patients. CONCLUSION: This study gives the first evidence that expressions of TLR2 and TLR4 in PBMCs were markedly elevated in LOAD patients.

Effects of the group II mGlu receptor agonist 2R,4R-APDC on dentate gyrus cell proliferation in the adult rat brain after diffuse brain injury.

OBJECTIVES: Diffuse brain injury (DBI) has been shown to increase the proliferation of granule cell precursors in the adult dentate gyrus (DG). However, the mechanism by which DBI-induced cell proliferation in the DG may enhance seizure susceptibility remains largely unknown. MATERIALS AND METHODS: Using bromodeoxyuridine (BrdU) immunohistochemistry, we examined the effects of group II metabotropic glutamate receptor (mGluR) agonist, 2R,4R-4-aminopyrrolidine-2,4-dicarboxylate (2R,4R-APDC), on cell proliferation in the DG after DBI. RESULTS: It has been found that 2R,4R-APDC significantly blocked DBI-induced increase in the number of BrdU-positive cells in the DG, especially in hilus. In addition, double-label immunofluorescence staining showed that treatment with APDC did not affect the differentiation of newborn cells into neurons or astrocytes. Taken together, our findings indicate that the activation of mGluR system may inhibit the DBI-induced cell proliferation in the DG, but not the differentiation of newborn cells. DISCUSSION: It is suggested that 2R,4R-APDC has potential neuroprotection via inhibiting the aberrant neurogenesis induced by DBI, which is an important pathological basis of seizure or other abnormalities following DBI.

Association of DAPK1 genetic variations with Alzheimer's disease in Han Chinese.

Apoptosis and autophagy are common physiological and pathological processes in the human body. Death-associated kinase protein 1 (DAPK1), which participates in the process of cell death, has attracted people's attention for its potential risk with late-onset Alzheimer's disease (LOAD). A recent study identified two single nucleotide polymorphisms (SNPs) in DAPK1 that show significant association with LOAD in Caucasians. In order to clarify the role of these genetic variations in Chinese population, we examined the genetic variations of DAPK1/rs4877365 and DAPK1/rs4878104 in a group of 400 LOAD patients and 400 healthy controls. All samples were recruited from Northern Han Chinese population. Data collected from this study showed that there were significant differences in genotype (P=0.02) and allele (P=0.007) frequencies of DAPK1/rs4878104 but not in DAPK1/rs4877365 between LOAD patients and controls. The "C" allele of rs4878104 worked as a protective factor of LOAD (P=0.0026, odds ratio/OR=0.75) in Han Chinese. Logistic regression analysis revealed that homozygosity was strongly associated with LOAD under a recessive model (P=0.003, OR=0.23). No significant association was observed between rs4877365 and LOAD. Haplotype analysis identified the G/T haplotype as a risk factor for LOAD (P=0.007, OR=1.33, 95% CI=1.08-1.64). This study provides the evidence that variation in DAPK1 gene influences susceptibility to LOAD in the Northern Han Chinese population.

Recovery of cognitive dysfunction in a case of delayed encephalopathy of carbon monoxide poisoning after treatment with donepezil hydrochloride.

Delayed encephalopathy following carbon monoxide poisoning is a serious complication. Here, we report a patient with delayed encephalopathy who suffered from cognitive disorders and urinary incontinence after a temporal normal period of 15 days after acute intoxication, and his cognitive function recovered gradually following donepezil hydrochloride treatment. Now, he can undertake slight farming work.

Alterations of NR2B and PSD-95 expression in hippocampus of kainic acid-exposed rats with behavioural deficits.

Temporal lobe epilepsy (TLE), characterized by spontaneous recurrent seizure (SRS), is associated with behavioural problems, but the underlying molecular mechanisms have not been clearly identified. In the present study, kainic acid (KA) was administered systemically in adult male Wistar rats to induce SRS. Behavioural performance analyses at 2, 4, and 6 weeks post-status epilepticus (post-SE) showed spatial learning memory deficit, anxiety and increased locomotor activity in rats with long-term SRS compared with rats without SRS after normal saline (NS) or KA-valproate (KA-VPA) treatment. No neuronal cell loss was observed in the hippocampus at 6 weeks post-SE. Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot analyses revealed that down-regulation of NMDA receptor subunit 2B (NR2B) and postsynaptic density protein-95 (PSD-95) expression in adult hippocampus was found at 4 weeks post-SE and a further decrease at 6 weeks post-SE compared with rats without SRS after NS or KA-VPA treatment. Furthermore, the decreased expression of NR2B and PSD-95 was correlated with the representatively behavioural deficit. These findings suggest that long-term SRS might decrease NR2B/PSD-95 expression in adult hippocampus and consequently cause behavioural deficits, including spatial learning memory deficit, anxiety and increased locomotor activity. Maintaining the expression of NR2B/PSD-95 might partially contribute to the normal behaviour in rats with long-term SRS.

Lipid peroxidation was involved in the memory impairment of carbon monoxide-induced delayed neuron damage.

Carbon monoxide (CO)-induced delayed neuron damage is the serious complication, but the underlying mechanisms are poorly understood. This study was designed to investigate the time-dependent changes of the lipid peroxidation (malondialdehyde, MDA) and antioxidative status (glutathione, GSH; glutathione peroxidase, GSH-Px; glutathione reductase, GR; and anti-reactive oxygen species anti-ROS) in nerve tissues for the possible mechanisms exploration. Adult rats were treated with CO by peritoneal injection, and sacrificed after day 0, 1, 3, 7, 14 and 21 of treatment. The results showed that the step-down latency progressively shortened while the numbers of error increased. Comparing with the level of day 0, MDA levels in serum, cerebral cortex and hippocampus significantly increased on day 1, 3 and 7. The level of GSH increased firstly but then decreased. The activities of GR, GSH-Px, and anti-ROS decreased in serum, cerebral cortex and hippocampus of rats after day 1, 3, 7, 14 and 21. Thus, we concluded that CO-mediated delayed neuron damage might be associated with elevation of lipid peroxidation and reduction of antioxidative status. The time-dependent changes of lipid peroxidation and antioxidative status in serum, cerebral cortex and hippocampus, at least in part, are involved in the toxic effects of CO poisoning on neuron.

Expressions of glutathione S-transferase alpha, mu, and pi in brains of medically intractable epileptic patients.

BACKGROUND: Glutathione S-transferases (GSTs) play an important role in metabolizing anti-epileptic drugs (AEDs) in liver. Expressions of GSTs in brain, which may result in poor efficacy of AEDs, have not been well studied. Using clinical cortex specimen from 32 intractable epileptic subjects and 8 non-epileptic controls, the present study investigated the correlation between GSTs and intractable epilepsy. RESULTS: Three different GST isoforms (alpha, mu, and pi) were detected with immunohistochemistry. GST-alpha expression was not seen in any cortex specimens. Sixty three percent (63%) of control and 53% of intractible epileptic specimens showed GST-mu immunoreactivity. No significant difference in intensity of GST-mu staining was observed between these two groups. GST-pi expression was found in endothelial cells and glial cells/astrocytes. Fifty percent (50%) of the control patients and 66% of the epileptic patients were GST-pi positive. The grading of epileptic patients was significantly higher than that of control patients (p < 0.01). CONCLUSION: High levels of GST-pi in endothelial cells and glial cells/astrocyte correlate to medical intractable epilepsy, suggesting that GST-pi contributes to resistance to AED treatment.

[Clinical study on effect of sanqi tongshu capsule in treating ischemic stroke: multicenter clinical trial].

OBJECTIVE: To observe the clinical effect and security of Sanqi Tongshu capsule in treating ischemic strkoe. METHOD: A multicenter, no dummy, open labeled clincal trail was conducted. 1 753 patient were enrolled in this clinical trial. All patients were treated with the Sanqi Tongshu capsule (200 mg, three times a day) for 28 days. The score of Europe Stroke Score (ESS) and Barthel Index (BI) were evaluated before and after treatment. The adverse reaction occurred in the therapeutic course was also observed. RESULT: Finally, 1742 cases and 1676 cases were respectively assessable by Full analysis set (FAS) analysis and Full analysis set (FAS) analysis. Clinical symptoms markedly improved in patients after treatment, and the score of ESS and BI had significantly improved (P<0.05). According to the score of ESS and BI, the total effect analysis indicated that the total effective rate in FAS and PPS were 81.86% and 84.43% respectively. The rate of adverse reaction was 3.44%, and the raction was so slight that didn't need to receive therapy or withdral drug. CONCLUSION: Sanqi Tongshu capsule is effect and secure in treating ischemic stroke without obvious adverse reaction.

2R, 4R-APDC decreases cell proliferation in the dentate gyrus of adult rats: the effect of 2R, 4R-APDC on cell proliferation.

This study investigated the effects of group II metabotropic glutamate receptor agonist, 2R, 4R-4-aminopyrrolidine-2,4-dicarboxylate (2R, 4R-APDC), on cell proliferation in the dentate gyrus of adult rats. 2R, 4R-APDC at a dose of 1 and 10 nmol/day resulted in decreased bromodeoxyuridine immunoreactive cells in the dentate gyrus. In addition, we found that APDC treatment had no effect on the number of BrdU+ and GFAP(+)-labeled cells or BrdU+ and NeuN(+)-labeled cells compared with controls. These data suggest that group II metabotropic glutamate receptor is an important site for glutamate's regulation on cell proliferation in the dentate gyrus, but 2R, 4R-APDC had no effects on newborn cell's ability to differentiate into neurons or astrocytes.

Poly(ADP-ribose) polymerase inhibitor is neuroprotective in epileptic rat via apoptosis-inducing factor and Akt signaling.

3-Aminobenzamide (3-AB), an inhibitor of poly(ADP-ribose) polymerase (PARP), has been proved to have neuroprotective properties. In this study, we examined the role of 3-AB in rat hippocampal neuron death induced by seizures. Our data showed that the seizures resulted in PARP activation and translocation of the apoptosis-inducing factor from the mitochondria to the nucleus, leading to neuron death. These effects could, however, all be abolished by 3-AB. Moreover, we showed that 3-AB facilitated Akt activation and decreased the activity of its downstream target, glycogen synthase kinase-3beta. Altogether, our data suggested that 3-AB might have a therapeutic value in seizure-induced hippocampal neuron damage, probably due to the inhibition of apoptosis and activation of Akt cell survival signaling.