OBJECTIVEThe investigated the effects of Dengzhan Xixin on brain water content, blood-brain barrier (BBB) permeability, T2-weighted imaging (T2WI), metabolites and the lesion ratio after cerebral ischemia-reperfusion injuries (IRI).

METHODThe 65 rats were randomly individed into three groups, the sham-operated group, the ischemia-reperfusion group and the Dengzhan Xixin treatment group. The models of ischemia-reperfusion of middle cerebral artery in rats were established by placing an intraluminal suture. The Dengzhan Xixin treatment group were injected 10% Dengzhan Xixin injection 22.5 mg kg(-1) after ischemia 1.5 h. The sh am-operated group (n=5) were sacrificed on 1 to measure brain water content and BBB permeability. The rats of the ischemia-reperfusion group (n=30) and the Dengzhan Xixin treatment group (n=30) were sacrificed at reperfusion for 6 h, 12 h, 1 d, 2 d, 4 d, 7 d, respectively, after ischemia 1.5 h. The additional 35 rats were individed into the same three groups. The changes of T2WI and metabolites in the brain were observed, and rats were sacrificed at reperfusion for 1 d, 2 d, 4 d after ischemia 1.5 h to determine the lesion ratio by TTC.

RESULTIn the ischemia-reperfusion group, brain water content(77.93+/-0.68)% and BBB permeability (3.77+/-0.28) increased after reperfusion for 6 h. The peak time of brain water content was at 4 d (83.82+/-0.49)% and BBB permeability was at 2 d (5.51+/-0.24)%. In the ischemia-reperfusion group and the Dengzhan Xixin treatment group, there were hyperintense signals in the injury region of T2WI. In the ischemia-reperfusion group after reperfusion for 1 d, the ratio of NAA/Cr decreased and Cho/Cr increased. In the Dengzhan Xixin treatment group, the ratio of NAA/Cr increased and Cho/Cr decreased. In the treatment group, the lesion ratio decreased by TTC was 16.78+/-1.45 and in the ischemia-reperfusion group was 21.27+/-1.73 at 2 d.

CONCLUSIONDengzhan Xixin may relieve cerebral ischemia-reperfusion injury by influencing the metabolites of brain, stabilizing BBB and decreasing brain edema.

Animals ; Blood-Brain Barrier ; drug effects ; metabolism ; Brain Ischemia ; complications ; metabolism ; pathology ; Flavonoids ; pharmacology ; Male ; Permeability ; drug effects ; Rats ; Rats, Wistar ; Reperfusion Injury ; complications ; metabolism ; pathology ; Water ; metabolism

Malignant glioma is the most frequent type in brain tumors. The prognosis of this tumor has not been significantly improved for the past decades and the average survival of patients is less than one year. Thus, an effective novel therapy is urgently needed. TNF-related apoptosis inducing ligand (TRAIL), known to have tumor cell-specific killing activity, has been investigated as a novel therapeutic for cancers. We have developed Ad-stTRAIL, an adenovirus delivering secretable trimeric TRAIL for gene therapy and demonstrated the potential to treat malignant gliomas. Currently, this Ad-stTRAIL gene therapy is under phase I clinical trial for malignant gliomas. Here, we report preclinical studies for Ad-stTRAIL carried out using rats. We delivered Ad-stTRAIL intracranially and determined its pharmacokinetics and biodistribution. Most Ad-stTRAIL remained in the delivered site and the relatively low number of viral genomes was detected in the opposite site of brain and cerebrospinal fluid. Similarly, only small portion of the viral particles injected was found in the blood plasma and major organs and tissues, probably due to the brain-blood barrier. Multiple administrations did not lead to accumulation of Ad-stTRAIL at the injection site and organs. Repeated delivery of Ad-stTRAIL did not show any serious side effects. Our data indicate that intracranially delivered Ad-stTRAIL is a safe approach, demonstrating the potential as a novel therapy for treating gliomas.
Adenoviridae/genetics ; Animals ; Blood-Brain Barrier ; Brain/drug effects/*metabolism/pathology ; Brain Neoplasms/genetics/metabolism/pathology/*therapy ; Clinical Trials, Phase I as Topic ; DNA, Viral/metabolism ; Disease Models, Animal ; Drug Delivery Systems ; Drug Evaluation, Preclinical ; *Gene Therapy ; Glioma/genetics/metabolism/pathology/*therapy ; Humans ; Liver/drug effects/metabolism/pathology ; Protein Multimerization/genetics ; Rats ; Spleen/drug effects/metabolism/pathology ; TNF-Related Apoptosis-Inducing Ligand/genetics/*pharmacokinetics

Malignant glioma is the most frequent type in brain tumors. The prognosis of this tumor has not been significantly improved for the past decades and the average survival of patients is less than one year. Thus, an effective novel therapy is urgently needed. TNF-related apoptosis inducing ligand (TRAIL), known to have tumor cell-specific killing activity, has been investigated as a novel therapeutic for cancers. We have developed Ad-stTRAIL, an adenovirus delivering secretable trimeric TRAIL for gene therapy and demonstrated the potential to treat malignant gliomas. Currently, this Ad-stTRAIL gene therapy is under phase I clinical trial for malignant gliomas. Here, we report preclinical studies for Ad-stTRAIL carried out using rats. We delivered Ad-stTRAIL intracranially and determined its pharmacokinetics and biodistribution. Most Ad-stTRAIL remained in the delivered site and the relatively low number of viral genomes was detected in the opposite site of brain and cerebrospinal fluid. Similarly, only small portion of the viral particles injected was found in the blood plasma and major organs and tissues, probably due to the brain-blood barrier. Multiple administrations did not lead to accumulation of Ad-stTRAIL at the injection site and organs. Repeated delivery of Ad-stTRAIL did not show any serious side effects. Our data indicate that intracranially delivered Ad-stTRAIL is a safe approach, demonstrating the potential as a novel therapy for treating gliomas.
Adenoviridae/genetics ; Animals ; Blood-Brain Barrier ; Brain/drug effects/*metabolism/pathology ; Brain Neoplasms/genetics/metabolism/pathology/*therapy ; Clinical Trials, Phase I as Topic ; DNA, Viral/metabolism ; Disease Models, Animal ; Drug Delivery Systems ; Drug Evaluation, Preclinical ; *Gene Therapy ; Glioma/genetics/metabolism/pathology/*therapy ; Humans ; Liver/drug effects/metabolism/pathology ; Protein Multimerization/genetics ; Rats ; Spleen/drug effects/metabolism/pathology ; TNF-Related Apoptosis-Inducing Ligand/genetics/*pharmacokinetics

To study the relationship between tau hyperphosphorylation and the function of glutamate transporter okadaic acid (OA), a protein phosphatase inhibitor, 20 ng in a 0.5 microl volume, was injected into the frontal cortex of rat brain and immunostaining was used to observe the phosphorylation of tau protein and the expression of excitatory amino acid transporter 1 (EAAT1) in the brain following the injection. The results showed that (1) the neurons in the center of the injection region displayed cytoplasmic shrinkage, swelling, nuclear pyknosis, and dislocation at the early stage, and necrosis appeared 3 d after the injection. However, most neurons in the peri-injected areas showed normal morphological characters with immuno positive reaction for AT8, a tau phosphorylated marker; (2) morphological analysis showed that tau hyperphosphorylation caused by OA treatment was mainly observed in the axons and dendrites of neuronal cells at 6 h in the cell body at 1 d, which brought about dystrophic neurites and neurofibrillary tangle (NFT)-like pathological changes; (3) the induction of glutamate transporter EAAT1 was observed in the involved areas corresponding to that with AT8 immunopositive staining, and the number of EAAT1-positive staining cells markedly increased at 12 h (P<0.01), peaked at 1 d (P<0.001), then decreased at 3 d following the injection. Combined with a confocal laser scanning microscopic analysis, double fluorescent immunostaining showed that EAAT1 positive staining appeared in neurons as well as astrocytes in the peri-injected areas of the frontal cortex. These results demonstrate that OA increases glutamate transporter EAAT1 expression in neurons while it induces tau hyperphosphorylation. However, the mechanism and significance of the induction of glutamate transporter EAAT1 expression remain to be further elucidated.
Animals ; Astrocytes ; drug effects ; metabolism ; Axons ; drug effects ; metabolism ; Brain ; cytology ; Dendrites ; drug effects ; metabolism ; Excitatory Amino Acid Transporter 1 ; metabolism ; Neurofibrillary Tangles ; pathology ; Neurons ; drug effects ; metabolism ; Okadaic Acid ; pharmacology ; Phosphorylation ; Rats ; tau Proteins ; metabolism

Animals ; Apoptosis ; drug effects ; Brain ; drug effects ; Caspase 3 ; metabolism ; Curcumin ; pharmacology ; Hypoxia-Ischemia, Brain ; metabolism ; pathology ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the toxicity of intragastrically administered N, N-dimethylformamide (DMF) in female Wistar rats, and to provide experimental data for the overall evaluation of DMF toxicity under different ways of exposure.

METHODSForty female Wistar rats weighing 150∼180 g were randomly divided into four groups: control group (treated with water) and three DMF exposure groups with doses of 50 mg/kg, 100 mg/kg, and 200 mg/kg. After oral administration of DMF once a day for 14 consecutive days, the rats were weighed and sacrificed. The liver, kidney, brain, and uterus were weighed to calculate organ indices. The pathological changes in the liver were examined by HE staining. The protein expression of HSP70 in the liver, kidney, and brain was determined. Finally, peripheral lymphocytes were collected from the arteria cruralis to determine DNA damage by comet assay.

RESULTSFourteen days after DMF exposure, the body weight and organ indices of the kidney, brain, and uterus showed no significant changes. However, the liver index showed concentration-dependent increase in all DMF exposed groups (3.52±0.21, 3.55±0.13, and 3.88±0.22 in the low-, medium-, and high-dose groups, respectively), as compared with the control group (3.24±0.28) (P < 0.05 or P < 0.01). The pathological damage in the liver also showed a concentration-dependent manner. Inflammatory cell infiltration and granular degeneration in centrilobular hepatocytes were observed in the high-dose group. No significant change in protein expression of HSP70 was observed in the liver, kidney, or brain of DMF-exposed rats (P > 0.05). DNA damage was induced by DMF, and the DNA percentage of lymphocyte comet tail, average tail length, and tail moment in exposed groups were all significantly increased as compared with the control group (P < 0.05 or P < 0.01).

CONCLUSIONGavaged DMF can induce liver injury and DNA damage in lymphocytes in rats 14 days after administration. There is no significant change in protein expression of HSP70 in the liver, brain, or kidney after DMF exposure.

Animals ; Brain ; drug effects ; pathology ; DNA Damage ; drug effects ; Dimethylformamide ; toxicity ; Female ; Gastric Lavage ; HSP70 Heat-Shock Proteins ; metabolism ; Kidney ; drug effects ; pathology ; Liver ; drug effects ; pathology ; Lymphocytes ; drug effects ; Rats ; Rats, Wistar ; Toxicity Tests

OBJECTIVETo investigate the effect of curcumin on the injury in hippocampal neurons and the expression of regulated upon activation nonnal T-cell expressed and secreted (RANTES) in hippocamp during cerebral ischemia/reperfusion (I/R) in rats with spontaneous hypertension (SH).

METHODSMale Wistar-Kyoto (WKY) rats and spontaneous hypertension rats (SHR) were randomly divided into five groups (n = 6): sham group (W-Sham and S-Sham group), ischemia/reperfusion group (W-/R and S/R group), curcumin group (S-Cur group) . Each group was splitted into 5 subgroups of 3 h,12 h, 1 d, 3 d and 7 d according to the time interval before reperfusion. Global brain ischemia/reperfusion model was established by 4-VO method. Hematoxylin-eosin staining (HE staining) was used to observe the vertebral cell morphology in hippocampal CA1 region. Nissl staining was applied to detect the average density of cone cells in hippocampal CA1 region. The expression of RANTES in hippocamp was determined by ELISA. The behavior of the rats was evaluated at 7 days after reperfusion. Results: Compared with the sham group rats, the ability of learning and memory was significantly decreased in ischemia/reperfusion group rats, the number of injured neurons were greatly elevated , the protein expression levels of RANTES was significantly increased (P < 0.05). Compared with W-I/R group rats, the ability of learning and memory in S-I/R group rats was greatly reduced, the number of injured neurons increased extremely, the protein expression level of RANTES was significantly enhanced( P <0.05). The number of injured neurons declined significantly in S-Cur group rats, the ability to learn and remember of these rats was improved and the RANTES protein content decreased significantly (P < 0.05).

CONCLUSIONSHR are more susceptible to ischemia/reperfusion induced hippocampal neuronal injury which may be improved by curcu min. Its underlying mechanism is possibly associated with the inhibition of RANTES protein expression level.

Animals ; Brain Ischemia ; metabolism ; pathology ; physiopathology ; Chemokine CCL5 ; metabolism ; Cognition ; drug effects ; Curcumin ; pharmacology ; Hippocampus ; cytology ; metabolism ; pathology ; Hypertension ; metabolism ; pathology ; physiopathology ; Male ; Neurons ; drug effects ; metabolism ; pathology ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Reperfusion Injury ; metabolism

OBJECTIVETo investigate the effect of Cedemex on cAMP and cGMP contents in different brain regions in morphine withdrawal rats precipitated by naloxone.

METHODA physical morphine dependent model of rats was established by subcutaneous injection of morphine in gradually increasing dosage within 7 days. cAMP and cGMP contents of VTA, cortex and hippocampus of the rat brains were determined by radioimmunoassay.

RESULTThe morphine withdrawal symptoms of rats were relieved significantly by ig Cedemex. Compared with the controls, cAMP content in the region of VTA, cortex and hippocampus of the morphine dependent rats were significantly higher (P < 0.05), while cGMP contents in those regions were significantly lower (P < 0.05). cAMP contents in the area of VTA, cortex and hippocampus of the morphine dependent rats were significantly reduced, while cGMP contents were significantly increased by ig Cedemex.

CONCLUSIONCedemex may significantly attenuate the morphine withdrawal symptoms in rats. The mechanism of this effect may be related to adjusting the contents of cAMP and cGMP in some brain regions.

Animals ; Brain ; drug effects ; metabolism ; pathology ; Cerebral Cortex ; drug effects ; metabolism ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Hippocampus ; drug effects ; metabolism ; Morphine ; adverse effects ; Rats ; Substance Withdrawal Syndrome ; metabolism

OBJECTIVETo observe the protective effect of verapamil pretreatment against cerebral ischemia-reperfusion injury in gerbils.

METHODSThirty-three Mongolian gerbils were randomized into the control group (group A, n=6, with sham operation), ischemia group (group B), and 3 verapamil groups (groups C, D, and E, n=7) with intraperitpneal verapamil injection (2 mg/kg) 48, 24 and 12 h before ischemia, respectively. In group A, the bilateral common carotid arteries were only exposed without clamping, and in the other 4 groups, the arteries were clamped for 20 min followed by reperfusion for 50 min. The gerbils were then decapitated and the forebrain cerebral cortex was removed to determine superoxide dismutase (SOD) and glutathione (GSH) activities and measure the contents of malondial dehyde (MDA), endothelin (ET) and calcitonin gene-related peptide (CGRP). The left forebrain cerebral cortex was sampled in each group to observe the ultrastructural changes under electron microscope.

RESULTSIn groups C and D, SOD activities were significantly higher than those in group B (P<0.05), and in group E, the SOD activity elevation was not statistically significant (P>0.05). In groups C, D and E, GSH activity was significantly higher than that in group B (P<0.05). MDA content was significantly lower in groups C and D than in group B (P<0.05), but comparable between groups E and B (P>0.05). ET content was also significantly lower in the pretreatment groups (P<0.05), but CGRP content higher (not statistically so, however) than those in group B. The more serious ultrastructural damage of the cerebral tissue was observed in group B, but only mild damage was found in the verapamil groups.

CONCLUSIONSVerapamil given 12-48 h before cerebral ischemia may protect the gerbils from cerebral ischemia-reperfusion injury by enhancing SOD, GSH activities and decreasing ET content.

Animals ; Brain ; drug effects ; metabolism ; pathology ; Brain Ischemia ; complications ; metabolism ; pathology ; prevention & control ; Endothelins ; metabolism ; Gerbillinae ; Glutathione ; metabolism ; Malondialdehyde ; metabolism ; Receptors, Calcitonin Gene-Related Peptide ; metabolism ; Reperfusion Injury ; complications ; metabolism ; pathology ; prevention & control ; Superoxide Dismutase ; metabolism ; Verapamil ; pharmacology

AIMTo investigate the changes of the brain NSE, S100 and ultrastructure and effect of ligustrazine in rats of chronic hypoxia and hypercapnia.

METHODSThirty rats were randomly divided into three groups: control group (A), hypoxia hypercapnia group (B), hypoxia hypercapnia added ligustrazine group (C). The brain NSE, S100 and ultrastructure were observed in rats using the technique of immunohistochemistry and electronic microscope.

RESULTS(1) The mPAP was significantly higher in rats of group B than that of group A and it was much lower in rats of group C than that of group B. Differences of mCAP were not significant in three groups. (2) Serum NO of group B was significantly lower than that of group A, Serum NO of group C was higher than that of group B. (3) Immunohistochemistry showed the average value of integral light density (LD) of NSE and S100 was significantly much lower in rats of group B than that of group A and it was higher in rats of group C than that of group B. (4) The neuron and astrocyte of group B showed vacuolar degeneration and the myelin sheath showed separate. Damage of neuron is alleviated in rats of group C.

CONCLUSIONThe hypoxia hypercapnia could induce damage of neuron and astrocyte in rats. The ligustrazine may be useful in protecting against hypoxia hypercapnia brain damage.

Animals ; Brain ; drug effects ; metabolism ; pathology ; Hypercapnia ; metabolism ; Hypoxia, Brain ; metabolism ; physiopathology ; Male ; Nitric Oxide ; blood ; Pyrazines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; S100 Proteins ; metabolism