• The Journal of Internal Korean Medicine
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• v.24 no.2
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• pp.181-189
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• 2003

The goal of this study is to investigate whether Uncariae Ramulus et Uncus can protect nerve cells against ischemic neuronal damage is caused by middle cerebral artery occlusion (MCAO) in rats' brains and to investigate whether the neuroprotective effect of Uncariae Ramulus et Uncus is concerned with IEGs(immediate early genes) expression. Uncariae Ramulus et Uncus(l00mg/kg) was administered immediately after 2 hours of MCAO and maintained for 7 days. On 7th days after 2 hours of MCAO, the brains of rats were sliced through the hippocampus. c-Fos immunohistochemistry stain and Cresyl violet stain were done for microscopic examination. Each number of viable neurons and c-Fos immunoreactive cells in CA1 was counted. The density of neurons and c-Fos immunoreactive cells were significantly decreased in MCAO-operated ischemic rats compared to that sham-operated rats. Administration of Uncariae Ramulus et Uncus group significantly elevated MCAO-induced decrease in density of neurons, and elevated MCAO-induced decrease in c-Fos immunoreactive cells. These results suggest that the neuroprotective effect of Uncariae Ramulus et Uncus against focal cerebral ischemia. Also, we hypothesized that neuroprotective mechanism of Uncariae Ramulus et Uncus is related to IEGs expression.

• The Korean Journal of Pharmacology
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• v.30 no.3
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• pp.273-289
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• 1994

Reversible brain ischemia was produced by occluding both common carotid arteries for 5 min, and the effects of aminoguanidine (AG), $DL-{\alpha}-difluoromethylornithine$ (DFMO), MK-801, and nimodipine (NM) on the ischemia induced changes of the polyamine, glutamate and acetylcholine levels in the hippocampus CA1 subfield and the specific $[^3H]\;MK-801$ binding to the hippocampus synaptosomal membranes were studied with a histological reference of the cresyl violet stained hippocampus. The basal putrescine level $(PT:\;74.4{\pm}8.8\;nM)$ showed a rapid increase (up to 1.7 fold) for 5 min of ischemia, remained significantly increased for 6 h, and then resumed the further increase to amount gradually up to about 3 fold 96 h after recirculation. However, the level of spermidine was little changed, and the spermine level showed a transient increase during ischemia followed by a sustained decrease to about 40% of the preischemic level after recirculation. The increase of PT level induced by brain ischemia was enhanced with AG or MK-801, but it was reduced by DFMO or NM. The basal glutamate level $(GT:\;0.90{\pm}0.l4\;{\mu}M)$ rapidly increased to a peak level of $8.19{\pm}1.14\;{\mu}M$ within 5 min after onset of the ischemia and then decreased to the preischemic level in about 25 min after recirculation. And NM reduced the ischemia induced increase of GT level by about 25%, but AG, DFMO and MK-801 did not affect the GT increase. The basal acetylcholine level $(ACh:\;118.0{\pm}10.5\;{\mu}M)$ did little change during/after brain ischemia and was little affected by AG or NM. But DFMO and MK-801, respectively, produced the moderate decrease of ACh level. The specific $[^3H]\;MK-801$ binding to the hippocampus synaptosomal membrane was little affected by brain ischemia for 5 min. The control value (78.9 fmole/mg protein) was moderately decreased by AG and MK-801, respectively but was little changed by DFMO or NM. The microscopic findings of the brains extirpated on day 7 after ischemia showed severe neuronal damage of the hippocampus, particularly CA1 subfield. NM and AG moderately attenuated the delayed neuronal damage, and DFMO, on the contrary, aggravated the ischemia induced damage. However, MK-801 did not protect the hippocampus from ischemic damage. These results suggest that unlike to the mode of anti-ischemic action of NM, AG might protect the hippocampus from ischemic injury as being negatively regulatory on the N-methyl-D-aspartate (NMDA) receptor function in the hippocampus.

• The Korean Journal of Pharmacology
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• v.32 no.3
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• pp.323-334
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• 1996

Male Mongolian gerbils $(60{\sim}80g)$ were given DL-difluoromethylornithine (DFMO; 250mg/kg, ip) and methylglyoxal bis(guanylhydrazone) (MGBG; 50 mg/k, ip), respectively, 1 h prior to transient (7 min) occlusion of bilateral common carotid arteries (OBC7) and a daily dose of one of them for 6 days after recirculation, and the polyamine contents, activities of ornithine and S-adenosylmethionine decarboxylases (ODC and SAM-DC), and light microscopic findings of the hippocampus were evaluated. The hippocampal putrescine (PT) levels of the control gerbils treated with saline (STGr), markedly increased after OBC7, showing a peak level at 24 h after recirculation. The peak PT level was reduced in DFMO treated gerbils (DTCr) and in MGBG treated gerbils (MTGr). And 7 days after recirculation, the PT level of DTGr was decreased to about 75% of the PT level in the sham operated group (nonTGr) and to about 55% of the STGr level, respectively. The hippocampal spermidine (SD) level of STGr tended to decline, showing the lowest value at 8 h after recirculation. But the spermidine (SD) level of DTGr was somewhat higher at 8 h after OBC7 than those of STGr and MTGr The hippocampal spermine (SM) levels of all the experimental groups were little changed for 7 days after OBC. OBC7 markedly increased the hippocampal ODC activity. reaching a maximum (about 3 times higher than preischemic level) at 8 h and rapidly recovered to the control value by 24 h in STGr gerbils, and the OBC7-induced increase of ODC activity was significantly attenuated by DFMO or MGBG treatment. Whereas OBC7 induced a rapid decrease of the hippocampal SAMDC activity follwed by gradual recovery to the preischemic level, and the decrease of the SAMDC activity was slightly attenuated by DFMO or MGBG treatment. 7 Days after OBC7 the histological finding of the hippocampal complex stained with cresyl violet showed an extensive delayed neuronal damage in the CA1 region and to a lesser extent, in the dentate gyrus, sparing the CA3 region. And the neuronal death was aggevated by DFMO but significantly attenuated by MGBG. The immunochemical reactivity of hippocampus to anti-GFAP antibody was significantly increased in the CA1 region and to a lesser extent, in the dentate gyrus 7 days after OBC7, but was little changed in the CA3. And the increase of the anti-GFAP immunoreactivity was moderately enhanced by DFMO and significantly suppressed by MGBG. These results suggest that the polyamine metabolism may play a modulatory role in the ischemic brain damage.

• Journal of Korean Medicine Rehabilitation
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• v.20 no.4
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• pp.1-15
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• 2010

Objectives : This study was performed to evaluate the effects of Hwangryunhaedok-tang(Huanglianjiedu-tang HHT) water extract on locomotor dysfunction induced by spinal cord injury(SCI) in rats. Methods : SCI was induced by mechanical contusion following laminectomy of 10th thoracic vertebra in Sprague-Dawley rats. HHT was orally given once a day for 14 days after SCI. Neurological behavior was examined with the Basso-Beattie-Bresnahan locomotor rating scale. Tissue damage and nerve fiber degeneration were examined with cresyl violet and luxol fast blue staining. Using immunohistochemisty, cellular damage to neurons and nerve fibers were examined against Bax and MAP-2. As inflammatory response markers, iNOS and COX-2 expressions were also examined. Results : 1. HHT ameliorated the locomotor dysfunction of the SCI-induced rats. 2. HHT attenuated the reduction of motor neurons in the ventral horn of the SCI-induced rat spinal cord. 3. HHT significantly reduced the number of Bax positive cells in the peri-lesion of the SCI-induced rat spinal cord. 4. HHT attenuated the reduction of MAP-2 positive cells in the peri-lesion of the SCI-induced rat spinal cord. 5. HHT significantly reduced the number of iNOS and COX-2 positive cells in the peri-lesion of the SCI-induced rat spinal cord. Conclusions : These results suggest that HHT improves the locomotor dysfunction of SCI by protecting motor neurons from cell death through anti-inflammatory effect.

• The Journal of Korean Medicine
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• v.35 no.4
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• pp.10-23
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• 2014

Objectives: This study evaluated the effects of Guibi-tang (GBT) on neuronal apoptosis and cognitive impairment induced by beta amyloid ($A{\beta}$), (1-42) injection in the hippocampus of ICR mice. Methods: $A{\beta}$ (1-42) was injected unilaterally into the lateral ventricle using a Hamilton syringe and micropump ($2{\mu}g/3{\mu}{\ell}$, $0.6{\mu}{\ell}/min$). Water extract of GBT was administered orally once a day (500 mg/kg) for 3 weeks after the $A{\beta}$ (1-42) injection. Acquisition of learning and retention of memory were tested using the Morris water maze. Neuronal damage and $A{\beta}$ accumulation in the hippocampus was observed using cresyl violet and Congo red staining. The anti-apoptotic effect of GBT was evaluated using TUNEL labeling in the hippocampus. Results: GBT significantly shortened the escape latencies during acquisition training trials. GBT significantly increased the number of target headings to the platform site, the swimming time spent in the target quadrant, and significantly shortened the time for the 1st target heading during the retention test trial. GBT significantly attenuated the reduction in thickness and number of CA1 neurons, and $A{\beta}$ accumulation in the hippocampus produced by $A{\beta}$ (1-42) injection. GBT significantly reduced the number of TUNEL-labeled neurons in the hippocampus. Conclusion: These results suggest that GBT improved cognitive impairment by reducing neuronal apoptosis and $A{\beta}$ accumulation in the hippocampus. GBT may be a beneficial herbal formulation in treating cognitive impairment including Alzheimer's disease.

• Journal of Acupuncture Research
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• v.21 no.2
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• pp.1-20
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• 2004

Objective : The aim of this study was to investigate effects of Angelica gigas Nakai(AGN) on the ischemic injury by intraluminal filament insertion in the rats. Methods : The ischemia was induced by intraluminal filament insertion into middle cerebral artery. AGN herbal acupuncture into SP10 was carried out during 3 weeks after ischemic injury. Eight-arm radial maze was designed for the behavioral task. AGN herbal acupuncture showed neuroprotective agents in cresyl violet, acetylcholinesterase(AchE), choline acetyltransferase(ChAT) and nerve growth factor(NGF)-stain. Then check the effect of regional cerebral blood flow(rCBF) according to AGN herbal acupuncture in rats. Results : The errors in the eight-arm radial maze task were significantly decreased in normal group compared with control group on 1~6days, AGN2(0.02g/kg) herbal acupuncture group on 1~5days, AGN3(0.1g/kg) on 1~3days, AGN4(0.5g/kg) on 1, 3~6days. The rate of correct choice was significantly increased in AGN1(0.01g/kg) and AGN4 herbal acupuncture groups. The density of neurons in the hippocampal CA1 was the most increased in normal group and AGN1, AGN3, AGN4 herbal acupuncture groups compared with control group. The density of AchE in the hippocampal CA1 had a tendency to increase in all the groups when they were compared with control group, but not significant. The density of ChAT in the hippocampal CA1 was significantly increased in normal group and AGN1, AGN4 herbal acupuncture groups compared with control group. The density of NGF in the hippocampal CA1 was significantly increased AGN4 herbal acupuncture group compared with control group. The rCBF was significantly increased in AGN1, AGN3 and AGN4 herbal acupuncture groups without the change of blood pressure. Conclusions : These results suggest that AGN herbal acupuncture can be used for controlling stroke in early stage as herbal medication.

• Korean Journal of Acupuncture
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• v.24 no.4
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• pp.99-110
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• 2007

Objectives : Epilepsy is one of the most common serious brain disorders that affect people of all ages, and it is characterized by recurrent unprovoked seizures. We examined whether acupuncture can reduce both the incidence of seizures and hippocampal cell death in dentate gyrus (DG) using a mouse model of kainic acid (KA)-induced epilepsy. Methods : ICR mice ($20{\sim}25$ g) were given acupuncture once a day at acupoint HT8 (sobu) bilaterally during 2 days before KA injection. After an intracerebroventricular injection of 0.1${\mu}g$ of KA, acupuncture treatment was subsequently administered once more (total 3 times), and the degree of seizure was observed for 20 min. Three hours after injection, we confirmed the neural cell death using cresyl violet staining and silver impregnation staining, and determined the expressions of c-Fos and glutamate decarboxylase (GAD)-67 using immunohistochemistry techniques in the DG. Results : KA induced epileptic seizure, neural cell death, increased c-Fos expression and decreased GAD-67 expression in the DG. Acupuncture treatment at HT8 reduced the severity of the epileptic seizure and inhibited neural cell death from KA. In addition, acupuncture normalized the expressions of c-Fos and GAD-67 in the same areas. Conclusions : These results demonstrated that acupuncture treatment at HT8 may reduce the KA-induced epileptic seizure and neural cell death in the DG possibly by normalizing c-Fos expressions and the gamma-aminobutyric acid neurons.

• The Journal of Internal Korean Medicine
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• v.31 no.4
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• pp.763-774
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• 2010

Objectives : In condition of brain infarction, irreversible axon damage occurs in central nerve system(CNS), because gliosis becomes physical and mechanical barrier to axonal regeneration. Reactive gliosis induced by ischemic injury such as middle cerebral artery occlusion is involved with up-regulation of GFAP and CD81. The current study is to examine the effect of the Uncariae Ramulus et Uncus on CD81 and GFAP expression in the rat brain following middle cerebral artery occlusion. Methods : In order to study ischemic injuries on brain, infarction was induced by middle cerebral artery occlusion(MCAO) using insertion of a single nylon thread, through the internal carotid artery, into a middle cerebral artery. Cresyl violet staining, cerebral infarction size measurement, immunohistochemistry and microscopic examination were used to detect the expression of CD81 and GFAP and the effect on the infarct size and pyramidal cell death in the brain of the rat with cerebral infarction induced by MCAO. Results : The following results were obtained 1. Measuring the size of cerebral infartion induced by MCAO in the rat after injection of Uncariae Ramulus et Uncus showed the size was decreased. 2. Intravenous injection of Uncariae Ramulus et Uncus showed pyramidal cell death protection in the hippocampus in the MCAO rat. 3. Water extract injection of Uncariae Ramulus et Uncus decreased GFAP expression significantly in the MCAO rat. 4. Uncariae Ramulus et Uncus water extract decreased CD81 expression in the MCAO rat. 5. The administration of water extract of Uncariae Ramulus et Uncus induced up-regulation of c-Fos expression significantly compared with MCAO. 6. The admistration of water extract of Uncariae Ramulus et Uncus increased ERK expression significantly compared with MCAO. Conclusion : We observed that Uncariae Ramulus et Uncus could suppress the reactive gliosis, which disturbs the axonal regeneration in the brain of the rat with cerebral infaction after MCAO by controlling the expression of CD81 and GFAP. The effect may be modulated by the up-regulation of c-Fos and ERK. These results suggest that Uncariae Ramulus et Uncus can be a candidate to regenerate CNS injury.

• Journal of Oriental Neuropsychiatry
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• v.18 no.1
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• pp.153-171
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• 2007

Objective : The aim of this study was to investigate effects of Gagambosim-Tang(GBT) of focal brain ischemic injury induced by intraluminal filament insertion in the rats. Method : The ischemia was induced by intraluminal filament insertion into middle cerebral artery Eight-arm radial maze was designed for the behavioral task. Gagambosim-Tang was orally administrated in SD rats for 21 days. The task was started on the 4th week after focal brain injury, and performed two trials per day for 6days. The effects of Gagambosim-Tang on neuroprotective agents in cresyl violet, choline acetyltranferase(ChAT), nerve growth factor (NGF)-stain and c-Fos with ischemic injury were investigated. Results : The errors in the eight-arm radial maze task were significantly decreased in normal group compared with control group on 2-6days, GBT lX(42.2 mg/ml)orally administrated group on 1days, GBT 6X(253.2 mg/ml) on 3, 5, 6days. The rate of correct choice was increased in normal and GBT 6X groups. The neuroprotective effect in the hippocampal CA1 was increased in normal and GBT 1X, GBT 6X groups compared with control group. The density of ChAT in the hippocampal CA1 was increased in normal and GBT6X groups compared with control group. The density of NGF in the hippocampal CA1 was increased normal and GBT6X groups compared with control group. The number of c-Fos-positive neurons in the hippocampal CA1 was increased in normal and GBT 6X groups compared with control group. Conclusion : These results suggest that Gagambosim-Tang may have protective effect on dementia.

• The Journal of Korean Medicine
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• v.26 no.2 s.62
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• pp.217-230
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• 2005

Objectives: Chungpaesagan-tang (CPSGT), which is frequently used for treating patients of cerebrovascular disease, has not been reported by clinical doctors concerning the effect of neuronal aptosis caused by brain ischemia. To study the effect of CPSGT on focal cerebral ischemia in normal and diabetic rats and SHR, focal cerebral ischemia was induced by transient MCAO, and after onset CPSGT was administrated. Methods: Rats (Sprague-Dawley) were divided into four groups: sham-operated group, MCA-occluded group, CPSGT­administrated group after MCA occlusion, and normal group. The MCA was occluded by intraluminal method. CPSGT was administrated orally twice (l and 4 hours) after middle cerebral artery occlusion. All groups were sacrificed at 24 hours after the surgery. The brain tissue Was stained with $2\%$ triphenyl tetrazolium chloride (TTC) or $1\%$ cresyl violet solution, to examine effect of CPSGT on ischemic brain tissue. The blood samples were obtained from the heart.~. Tumor necrosis $factor-\alpha$ level and interleukin-6 level of serum was measured from sera using enzyme-linked immunoabsorbent assay (ELISA). Then changes of immunohistochemical expression of $TNF-\alpha$ in ischemic damaged areas were observed. Results: In NC+MCAO+CP and DM+MCAO+CP, CPSGT significantly (p<0.01) decreased the number of neuron cells compared to the control group. CPSGT markedly reduced (p<0.01) the infarct size of the forebrain in distance from the interaural line on cerebral ischemia in diabetic rats. CPSGT significantly reduced the $TNF-\alpha$ expression in penumbra region of damaged hemisphere in diabetic rats. Conclusions: CPSGT had a protective effect on cerebral ischemia in SD rats, especially in diabetic rats compared with normal SD rats.