• The Korea Journal of Herbology
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• v.27 no.2
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• pp.61-67
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• 2012

Objective : Angelica Gigas Nakai is a popular oriental medicine used for the treatment of vascular diseases. The aim of this study is to investigate neuroprotective effect of the water extract of Anelicae Gigantis Radix Palva (AG) in transient middle cerebral artery occlusion (tMCAO)-induced ischemic rats via the regulation of angiogenesis-related molecules. Methods : Sprague-Dawley rats were intraperitoneally administrated with AG water extract at doses of 10, 25, 50 mg/kg body weight after tMCAO (90 min occlusion). reperfusion for 24 hr infarction volumes were measured by 2,3,5-tetrazolium chloride (TTC) staining. Brain tissues were observed neuronal cell injuries by nissl staining, and also brain-blood barrier (BBB) permeability change by evans blue. Expression of vascular endothelial growth factor (VEGF), angiopoietin-1 (Ang-1) and Tie-2 receptor protein in brain tissues was determined by western blot. Results : AG water extract significantly reduced infarction volume in ischemic brains of rats, degradation of neuronal cell, BBB permeability and expression of VEGF protein dose-dependently. Ang-1 protein was increased dose-dependantly, not significantly. Conclusion : This study suggests that AG water extract shows neuroprotective effect by preventing BBB breakdown, with regulating angiogenesis factor VEGF and Ang-1.

• Korean Journal of Head & Neck Oncology
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• v.9 no.1
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• pp.3-9
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• 1993

The benficial effects for perfusion in the preservation of free flaps have been controversial in the clinical and experimental field until now. This study was undertaken to observe the effect of UW solution. a recently developed. high molecular weight. organ perfusion solution. for protection of ischemic injury in normothermic free myocutaneous flaps. Forty rabbits were used in this sutdy. A 1x2x1cm sized gastrocnemius myocutaneous flap based on the feeding vessel from common femoral artery was made. The author set up the ischemic time for 12 hours in these flaps. The flap was washed out with normal saline(control grop, n=10), urokinase(comparative group I, n=10), UW solution before ischemic time(comparative group II, n=10) and UW solution before ischemic time and pentoxifylline before reperfusion(comparative group III, n=10). Afterthen, reperfusion was made for 12 hours. After this procedure, we checked the degree of ischemia and necrosis of myocutaneous flap by gross finding, electrical stimulation test of muscle, triphenyltetrazolium chloride staining and wet/dry weight ratio. The degree of necrosis of comparative group II and III were lesser than control and urokinase group in gross finding(p<0.05). In the electrical stimulation test of muscle, there was no statistical difference between control($1.76{\pm}1.01$) and urokinase($2.36{\pm}\1.02$) group however the muscular power of comparative group II($3.54{\pm}0.93$) and III($3.49{\pm}1.37gm/mm^2$) demonstrated significantly higher than control group(p<0.05). The ischemic findings were found in seven cases of control group and three cases of urokinase group but there were no ischemic findings in comparative group II and III in TIC stain(p<0.05). In the wet/dry weight ratio of flaps in order to evaluate the tissue edema. there was no statistical difference between control($4.55{\pm}0.29$) and III($3.75{\pm}0.48$) were scored significantly lesser than control and urokinase group (p<0.05). These results suggest that perfusion washout with UW solution improves the viability of normothermic free myocutaneous flap by inhibition of cellular swelling.

• Journal of Sasang Constitutional Medicine
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• v.26 no.2
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• pp.165-179
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• 2014

Objectives To evaluate the neuroprotective effects of Hyangsayangwi-tang (HY), a Korean traditional medicinal prescription in a Parkinson's disease mouse model. Methods Four groups(each of 10 mouse per group) were used in this study. The neuroprotective effect of HY was examined in a Parkinson's disease mouse model. C57BL/6 mouse treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP, 30mg/kg/day), intraperitoneal (i.p.) for 5 days. Slow behavioral responses and memory disorder is the major clinical symptoms of PD. In order to investigate the effect of HY on recovery of behavioral deficits and memory, we examined the motor function and memory by using Morris water maze and Forced swimming test. Ischemic mouse brain stained with TTC(2,3,5 triphenyl tetrazolium chloride) in the MPTP-induced Parkinson's disease to find out ischemia and tissue damage in mouse. The convenient, simple, and accurate high-performance liquid chromatography (HPLC) method was established for simultaneous determination of neurotransmitters in MPTP-HY group. To measure the amount of dopamine in mice brain, striatum-substantia nigra, was examined by Bradford assay. Immunohistochemistry was examined in the MPTP-induced Parkinson's disease (PD) mouse to evaluate the neuroprotective effects of Hyangsayangwi-tang on hippocampal lesion, ST and SNpc. Results and Conclusions Hyangsayangwi-tang (HY) prevents MPTP-induced loss of serotonin, hippocampus and TH-ir cell.

• The Korea Journal of Herbology
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• v.29 no.6
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• pp.85-93
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• 2014

Objectives : We compared with the effects of different parts (root head, root body and hairy root) of Angelica gigas Nakai (Angelicae Gigantis Radix, AG) with on middle cerebral artery occlusion(MCAO)-induced ischemic rats, and on LPS-induced inflammatory response in BV2 microglia. Methods : The 30% ethanol and water extracts of different parts of AG were prepared. Each extract (50 and 100 mg/kg) was administrated intraperitoneally once in MCAO-induced ischemic rats. We measured infarction volumes by TTC staining, and investigated the expression of iNOS, Bax, Bcl-2 and caspase-3 by Western blot. BV2 cells were treated with each extract for 30 min, and then stimulated with LPS. The levels of NO was measured by Griess assay. The expression of iNOS, Cox-2 and proinflammatory cytokines ($TNF-{\alpha}$, $IL-1{\beta}$, and IL-6) were determined RT-PCR and Western blot. The phosphorylation of ERK1/2 and JNK MAPK was determined by Western blot. Results : Among different parts of AG, the 30% ethanol and water extracts of hairy root significantly decreased infarction volume in ischemic brains and inhibited the expression of iNOS, bax and caspase-3. The extracts of hairy root significantly inhibited LPS-induced production of NO, $TNF-{\alpha}$ and IL-6 in BV2 cells, and suppressed the expression of iNOS and COX-2. The hairy root extracts attenuated LPS-induced phosphorylation of ERK1/2 and JNK MAPK in BV2 cells. Conclusions : Our results indicate that the root hairy of AG has a good neuroprotective and anti-inflammatory effects in ischemic stroke compared to other parts.

• The Korean Journal of Food And Nutrition
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• v.28 no.6
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• pp.965-972
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• 2015

This study was conducted to evaluate the neuroprotective effects of Cheonggukjang extract in in-vitro and in-vivo models. T98G-human glioblastoma cells were pretreated with various concentrations (1~10 mg/mL) of Cheonggukjang extract for 24 h and then exposed to $H_2O_2$ (1 mM) for 3 h. The neuroprotective effects of Cheonggukjang extract were measured using a CCK-8 kit assay, total antioxidant capacity (TAC) assay, reactive oxygen species (ROS) assay, and lactate dehydrogenase (LDH) release assay. The early stage focal ischemia rodent model was used as the in-vivo neurotoxicity model. Various concentrations (10~200 mg) of Cheonggukjang extract were administered to the animal models for 1 week. Peripheral blood was analyzed for glutathione peroxidase (GPx) expression by ELISA, and infarct volume reduction was analyzed by TTC staining. Cheonggukjang extract significantly (p<0.05) increased cell viability in T98G cells against $H_2O_2$ as well as against the induced neurotoxicity. Indeed, treatment with the Cheonggukjang extract induced a decrease in ROS and LDH expression and increased TAC significantly (p<0.05). However, Cheonggukjang extract did not induce a decrease in infarct volume or an increase in GPx expression in the in-vivo model. Despite the limitation in neuroprotection, Cheonggukjang extract may be useful for treating ROS injury.

• The Korea Journal of Herbology
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• v.27 no.2
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• pp.77-83
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• 2012

Objectives : This work was designed to investigate the effect of The root of Achyranthes japonica Nakai (AJN) water extract on serum deprivation reperfusion-induced apoptosis in PC-12 cells and transient middle cerebral artery occlusion (tMCAO)-induced ischemic brains of rats. Methods : Apoptosis in PC12 cells was induced by serum deprivation and reperfusion. The cells were treated with AJN water extract at doses of 0.5 and 1.0 mg/ml for 24 hr after inducing the apoptosis. Cell viability was determined by WST-1 assay. The expression of caspase-3 protein was determined by Western blot. Ischemic brains were prepared from tMCAO-induced ischemic rats after oral administration with AJN at dose of 50 and 100 mg/kg, and then brain infarction was measured by TTC staining. Results : AJN significantly increased the cell viability in apoptocic-induced PC-12 cells, and also decreased the expression of caspase-3 protein. Furthermore, the administration of AJN significantly inhibited tMCAO-induced brain infarction in rats. Conclusions : Our results suggest that AJN extract has a neuroprotective property via suppressing the apoptosis in PC12 cells and the infarction of ischemic brains.

• Journal of Society of Preventive Korean Medicine
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• v.13 no.1
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• pp.13-27
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• 2009

This study aimed to validate neuroprotective effect of Chungpaesagan-tang on the early stage of cerebral ischemic damage. Cerebral ischemic damage was induced by the middle cerebral artery occlusion (MCAO) for 2 hours in the Sprague-Dawley rats. Water extract of Chungpaesagan-tang(8.7g/kg) was administered orally twice at 1 and 4 hours after the MCAO. Neurological score was tested at 3 and 24 hours after the MCAO and Chungpaesagan-tang administration. At 24 hours after the MCAO, infarct volume and edema ratio was evaluated with the TTC staining. Apoptotic cell death in cerebral cortex and caudate putamen was observed with cresyl violet staining and TUNEL labeling. Bax expression in the MCAO rat brain was stained with immunohistochemistry. Chungpaesagan-tang improved neurological and behavioral impairment of the MCAO rats and reduced infarct area, infarct volume and brain edema formation. Chungpaesagan-tang attenuated cell death percentage in cortex penumbra and reduced TUNEL positive cells in cortex penumbra and in caudate putamen of the MCAO rats. Chungpaesagan-tang reduced Bax positive neurons in caudate putamen and reduced c-Fos positive neurons in cortex penumbra of the MCAO rats. Chungpaesagan-tang intensified neuronal HSP72 expression in cortex penumbra of the MCAO rats. In results, Chunpaesagan-tang reduces infarct volume and edema formation through anti-apoptotic effect. This result suggests that Chunapaesagan-tang has an adequate neuroprotective effect on the early stage of cerebral ischemic damage.

• YAKHAK HOEJI
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• v.41 no.1
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• pp.117-125
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• 1997

The pharmacological effects of benzopyran potassium channel openers (lemakalim, KR-30450 and KR-30818) on the occlusion/reperfusion-induced myocardial infarction were investigat ed. In anesthetized rats, subjected to 45-min occlusion of the left anterior descending coronary artery (LAD) followed by 90-min reperfusion, the infarct size was measured by calculating the ratio of infarct zone to area at risk (IZ/AAR) with the Evans blue/TTC technique. Rats were intravenously given vehicle (1% DMSO), lemakalim, KR-30450, and KR-30818 alone or in combination with a selective K$_{ATP}$ blacker glibenclamide, 30 min prior to coronary occlusion. Compared to vehicle, lemakalim (30 ${\mu}$g/kg i.v.), the active enantiomer of cromakalim, had a tendancy to decrease infarct size. KR-30450(30 ${\mu}$g/kg, i.v.). the newly synthetized potassium channel openers (PCOs), caused a reduction of infarct size (from 70${\pm}$4%to 57${\pm}$5%). but KR-30818 (30 ${\mu}$g/kg, i.v.), a metabolite of KR-30450. did not modify infarct size. It seem ed likely that glibenclamide (0.3mg/kg, i.v.), given in combination, reduced the effects of these PCOs, especially KR-30450 (30 ${\mu}$g/kg, i.v.) on the infarct size. These results indicate that. in the coronary occluded rat model of ischemia, lemakalim and KR-30450 may exert cardioprotective activity through a reduction of infarct size, the effect being considered related to the opening of K$_{ATP}$ channel.

• The Journal of Korean Medicine
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• v.21 no.2
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• pp.25-36
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• 2000

Objectives : This study demonstrates the effects of Daejo-hwan on learning and memory impairment induced by L-NAME (75 mg/kg) treatment and on cerebral ischemic damage induced by middle cerebral artery (MCA) occlusion in rats. Methods : Daejo-hwan emulsion (73.3 mg/100 g/l ml) was administered to rats along a timed study schedule. The Moms water maze was used for learning and memory test of the rats. The MCA was occluded by using the intraluminal thread method. The brain slices were stained by 2 % triphenyl tetrazolium chloride (TTC) and 1 % cresyl violet solution. Infarct size, neuron cell number and size in penumbra was measured by using computer image analysis system. Results : 1. The escape latency of the Daejo-hwan treated group decreased significantly with respect to the control group. 2.The memory score of the Daejo-hwan treated group showed increase tendency, And the swimming distance was not different between the normal, the control, and the Daejo-hwan treated group. 3. The infarct size of the Daejo-hwan treated group decreased significantly with respect to the control group. 4. The total infarct volume of the Daejo-hwan treated group showed decrease tendency. And the brain edema index of the Daejo-hwan treated group decreased significantly with respect to the control group. 5. The neuron cell number and cell size in penumbra of the Daejo-hwan treated group increased significantly with respect to the control group. Conclusions : According to the above results, it is supposed that Daejo-hwan is clinically applicable to the vascular dementia.

• The Journal of Korean Medicine
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• v.21 no.2
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• pp.68-78
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• 2000

Objective : This research was performed to investigate protective effects of Sophora subprostrata, against ischemic brain damage after a middle cerebral artery(MCA) occlusion. The effect was estimated using histological test, neurobehavioural test, and biochemical test. Methods : Rats(Sprague-Dawley) were divided into four groups: Sham operated group, MCA occluded group, Sophora subprostrata administrated group after MCA occlusion, and Normal group. The MCA was occluded by intraluminal method. Sophora subprostrata was administrated orally twice(l and 4 hours) after middle cerebral artery occlusion. The neurobeavioural test was performed at 3 hours, 6 hours, 9 hours and 24 hours after the surgery by posture reflex test and swimming behavioural test. All groups were sacrificed at 24 hours after the surgery. The brain tissue was stained with 2% triphenyl tetrazolium chioride(TTC) or 1 % cresyl violet solution, to examine effect of Sophora subprostrata on ischemic brain tissue. The blood samples were obtained from the heart of rats. Tumor necrosis factor-a level was measured from sera using Enzyme-Linked Immunoabsorbent Assay(ELISA). Results : The results showed that (1) Sophora subprostrata reduced infarct size and total infarct volume by 54.8% compared to the control group, (2) that neuronal death, which was shown by decrease in cell number and size, was attenuated significantly in the boundary area of the infarction, (3) that serum $TNF-{\alpha}$ㆍlevel was reduced significantly, and finally, there was significant recovery of motor deficit at 3 hours after MCA occluded by Swimming behavioural test. Conclusions :In conclusion, Sophora subprostrata has protective effects against ischemic brain damage at the early stage of ischemia.