• 한국콘텐츠학회논문지
• /
• 제19권7호
• /
• pp.186-191
• /
• 2019

서론: 최근 노년층에서 뇌졸중 발병률이 증가하고 있다. 현재 뇌졸중 치료제 및 방법이 많이 개발되어 있으나 치료 후에도 후유증 등이 많이 남게 된다. 그래서 아직도 많은 과학자나 임상 의사들이 이를 치료하기 위한 약물 및 방법을 연구하고 있는 실정이다. 많은 연구 중 뇌졸중 치료 연구를 위한 표준화된 실험 동물연구는 드물며, 표준화된 Nylon thread를 이용한 중대뇌동맥 폐쇄모델(MCAO, middle cerebral artery occlusion)의 성공률에 대한 연구는 거의 없다 방법: 본연구는 $0.18{\pm}0.02mm$의 지름을 가진 5-0 Nylon thread를 중대뇌동맥에 삽입하였다. 60분 동안 삽입한 후에 봉합해 놓았던 부위를 다시 절개하여 Nylon thread를 빼내고, 막았던 혈관의 매듭을 풀어주어, 다시 혈액이 공급되게 하였다. 그로부터 23시간 후에 뇌를 내어 1mm 두께로 자른 후 1.5% TTC(2',3',5'-triphenyl-tetrazolium chloride)로 15분간 염색하고, 4% PFA(paraformaldehyde)로 15분 동안 고정하였다. 결과: Nylon thread를 삽입하여, MCA occlusion 50마리, ICA occlusion 14마리, 제대로 된 MCAO model보다 좀 더 깊게 들어간 distal MCAO model 36마리, 너무 깊은 MCA나 ACA까지 들어가서 상보적인 괴사를 나타내는 occlusion model 1마리, 그리고 경색이 일어나지 않은 마우스 50마리를 확인하였다. 결론: 이에 본 연구에서는 Nylon Thread를 생쥐의 무게에 따라 32~36g 인 생쥐는 9mm로 삽입하여주고, 37~40g인 생쥐는 9mm+0.5mm의 깊이로 삽입하여서 1hr의 occlusion과 23hr의 reperfusion을 주어 생쥐를 TTC 염색을 통하여 괴사가 일어난 부분을 확인하였고, 생쥐에서 가역적인 뇌혈관 경색으로 151말중 101마리에서 뇌경색을 유도 할 수 있었다(66.9%).

• 대한한방내과학회지
• /
• 제30권4호
• /
• pp.674-684
• /
• 2009

Objectives : In conditions of brain infarction, irreversible axon damage occurs in the central nerve system (CNS), because gliosis becomes a physical and a 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. This study was undertaken to examine the effect of the Gongjin-dan (GJD) on CD81 and GFAP expression and its pathway in the rat brain following middle cerebral artery occlusion (MCAO). Methods : In order to study ischemic injuries on the brain, infarction was induced by 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. Also, c-Fos and ERK expression were measured to investigate the signaling pathway after GJD administration in MCAO rats. Results : Measuring the size of cerebral infarction induced by MCAO in the rat after injection of GJD showed the size had decreased. GJD administration showed pyramidal cell death protection in the hippocampus in the MCAO rat. GJD administration decreased GF AP expression in the MCAO rat. GJD administration decreased CD81 expression in the MCAO rat. GJD administration induced up-regulation of c-FOS expression compared with MCAO. GJD administration induced down-regulation of ERK expression compared with MCAO. Conclusion : We observed that GJD could suppress the reactive gliosis, which disturbs the axonal regeneration in the brain of a rat with cerebral infarction after MCAO by controlling the expression of CD81 and GFAP. The effect may be modulated by the regulation of c-Fos and ERK. These results suggest that GJD can be a candidate to regenerate CNS injury.

• Journal of Acupuncture Research
• /
• 제36권4호
• /
• pp.220-229
• /
• 2019

Background: The therapeutic potential of Bee Venom Pharmacopuncture (BVP) on acute ischemic cerebral infraction was determined in mice in vivo and in vitro. Methods: Analysis of acute ischemic cerebral infraction was performed using 7 week old male ICR mice (n = 20) and microglial BV-2 cells. Bee venom ($5{\mu}g/kg$) was injected into the caudal vein of middle cerebral artery occlusion (MCAo) mice (1 hour after reperfusion, 3 hours after MCAo probe insertion), and also used to treat LPS-stimulated microglial BV-2 cells (1, 2, $5{\mu}g/mL$). Markers of inflammation were monitored. Results: NO declined statistically significantly in BVP treated MCAo mice compared to the untreated MCAo group (p < 0.05). Compared to the MCAo group, the BVP-treated MCAo group showed a decreased production volume of malondialdehyde, but an increased glutathione/oxidized glutathione ratio. Compared to the untreated MCAo group, the BVP treated MCAo group showed a statistically significant decline in TNF and $IL-1{\beta}$ levels (p < 0.05). BVP inhibited the levels of p65, p50, $p-I{\kappa}B-{\alpha}$, and levels of p-ERK1/2, p-JNK2, p-P38 declined. Conclusion: BVP is effective at dampening the inflammatory response in vivo and in vitro and may supplement rt-PA treatment.

• 대한한의학회지
• /
• 제20권4호
• /
• pp.82-92
• /
• 2000

The purpose of the present study was to explore the proper method for animal stroke model of Oriental medicine To this end, brain ischemia was induced by distal middle cerebral artery occlusion(dMCAO) and proximal middle cerebral artery occlusion(pMCAO) and evaluated with the method of Triphenyl Tetrazolium Chloride (TTC) staining and Swimming Behavior Test. Results demonstrated that first, infarct size and volume of pMCAO group were significantly bigger that those of dMCAO group. Second, analysis of swimming behavior test revealed that the percentage of left turning angles of pMCAO was significantly bigger than that of dMCAO. Third, during swimming behavior test, there were peculiar traces of small successive circles that represent motor dysfunction and conscious disturbance among dMCAO group. The results of the study thus indicate that non-invasive intraluminal method of pMCAO was the appropriate animal stroke model for Oriental medicine in the light of brain ischemia as hemiplesia and conscious disturbance.

• 대한한의학회지
• /
• 제30권2호
• /
• pp.117-126
• /
• 2009

Objectives: This study was performed to investigate the effects of Semen Persicae (SP) on infarct volume, COX-2 protein expression in the middle cerebral artery occlusion (MCAo) rats. Methods: Twenty-eight rats were randomly assigned to four groups (MCAo experimental group, MCAo control group, sham experimental group, control group). The middle cerebral artery (MCA) was occluded in the MCAo group by proximal focal cerebral ischemia rat model, while the MCA was not occluded in the sham group. SP extraction was administrated for 4 days to each experimental group. Neuroprotective effects were investigated by measurement of brain damage using 2, 3, 5-triphenyltetrazolium chloride staining and analysis of COX-2 protein expression by western blotting. Results: The occurrence of infarct volume in the SP oral administration group decreased compared to the control group. COX-2 protein expression in the SP oral administration group decreased compared to the control group. Conclusions: The present study demonstrates the effect of SP in reducing infarct volume and decreasing COX-2 expression.

• 대한한의학회지
• /
• 제25권1호
• /
• pp.117-128
• /
• 2004

Objectives : This research was performed to investigate effect of Samul-tang-gamibang against focal cerebral ischemic damage after middle cerebral artery occlusion(MCAO). Methods : This research was used rats which were against focal cerebral ischemic damage by MCAO. It was used Zea Longa's theory and Belayev's methods to give rise to focal cerebral ischemic damage by MCAO. After 7days later, we drew out the brain and then had frozen and dyeing it and we had taken a picture to measure of the damaged area in each brain section. We determined the Neurological Index and tested the Foot-fault test and Roatated test to appraise the fall of motion ability result from cerebral ischemic damage. Results : The results of the experiment are as follows. 1. Samul-tang-gamibang reduced infarct size of sample group compared to control group at 7 day after MCAO. 2. Samul-tang-gamibang reduced infarct volume of sample group compared to control group at 7 day after MCAO. 3. Samul-tang-gamibang reduced foot-fault index of sample group compared to control group at 5,7 day after MCAO. Conclusions : Samul-tang-gamibang has protective effects against ischemic brain damage and had significant reduced infarct size and infarct volume of Rt-MCAO.

• Biomolecules & Therapeutics
• /
• 제8권2호
• /
• pp.160-166
• /
• 2000

The present study examined influence of various ischemic duration on extent of focal ischemic brain injury induced by middle cerebral artery occlusion (MCAO) in rats. The MCAO was produced by insertion of a 17 mm silicone-coated 4-0 nylon surgical thread to the origin of MCA through the internal carotid artery for 30, 60, 90, 120 min (transient) or 24 hr (permanent) in male Sprague-Dawley rats under isoflurane anesthesia. Reperfusion in transient MCAO models was achieved by pulling the thread out of the internal carotid artery. Only rats showing neurological deficits characterized by left hemiparesis and/or circling to the left, were included in cerebral ischemic groups. The rats were sacrificed 24 hr after MCAO and seven serial coronal slices of the brain were stained with 2,3,5-triphenyltetrazolium chloride. Infarct size was measured using a computerized image analyzer. Ischemic damage was common in the frontoparietal cortex (somatosensory area) and the lateral segment of the striatum while damage to the medial segment of the striatum depended on the duration of the occlusion. In the 30-min MCAO grouts, however, infarcted region was primarily confined to the striatum and it was difficult to clearly delineate the region since there was mixed population of live and dead cells in the nucleus. Infarct volume was generally increased depending on the duration of MCAO, showing the most severe damage in the permanent MCAO group. However, there was no significant difference in infarct size between the 90-min and 120-min MCAO groups. % Edema also tended to increase depending on the duration of MCAO. The results suggest that the various focal ischemic rat models established in the present study can be used to evaluate in vivo neuroprotective activities of candidate compounds or to elucidate pathophysiological mechanisms of ischemic neuronal cell death.

• 대한한방내과학회지
• /
• 제31권4호
• /
• pp.763-774
• /
• 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.

• 대한약침학회지
• /
• 제14권3호
• /
• pp.5-17
• /
• 2011

Objectives : Gliosis becomes physical and mechanical barrier to axonal regeneration. Reactive gliosis induced by middle cerebral artery occlusion is involved with up-regulation of CD81 and GFAP (Glial fibrillary acidic protein). The current study is to examine the effect of the Angelica gigas Nakai(intravenous injection. 100 mg/kg twice in a day) on CD81 and GFAP of the rat in the brain after middle cerebral artery occlusion. Methods : Cerebral infarction was induced by middle cerebral artery occlusion. And after intravenous injection of water extract of Angelica gigas Nakai, the size of cerebral infarction was measured. Examination of optical microscope were also used to detect the expression of CD81 and GFAP in the brain of the rat. Results : The following results were obtained : We found that size of cerebral infarcion induced by MCAO (Middle Cerebral Artery Occlusion) in rats were decreased after intravenous injection of Angelica gigas Nakai. We injected the extract of Angelica gigas Nakai to the MCAO in rats, and the optical microscope study showed that Angelica gigas Nakai had effect on protecting the cells of hippocampus. We found that GFAP, CD81 and ERK of the brain in rats with cerebral infarction after MCAO were meaningfully decreased after intravenous injecting Angelica gigas Nakai. We found that c-Fos expression of the brain in rats with cerebral infarction after MCAO were significantly increased after intravenous injecting Angelica gigas Nakai. Conclusions : These results indicate that Angelica gigas Nakai could suppress the reactive gliosis, which disturbs the astrocyte regeneration in the brain of the rat with cerebral infarction after MCAO by controlling the expression of CD81 and GFAP. And the effect may be modulated by the up-regulation of c-Fos and ERK.

• 대한한의학회지
• /
• 제27권2호
• /
• pp.70-85
• /
• 2006

Objectives; This study examined the neuroprotective effect of Hyulbuchookautang (血府逐瘀湯, HBCAT)against neural damage following focal cerebral infarction. Methods : Sprague-Dawley Rats were induced with focal cerebral infarction by temporal middle cerebral artery occlusion (MCAO). The rats were divided into 2 groups. We treated extract of HBCAT to one group after operation (sample group), and the other group wasn't treated after operation (control group). We observed neurological scores and TIC-stained infarct area, total infarct volume in brain sections and Bax-positive neurons, HSP70- positive neurons in brain regions. Results : HBCAT treatment at 3 days after MCAO reduced neurological scores induced by MCAO. HBCAT treatment at 5 days after MCAO reduced TTC-stained infarct area in brain sections induced by MCAO. HBCAT treatment at 5 days after MCAO reduced total infarct volume in brain sections induced by MCAO. HBCAT treatment after MCAO reduced Bax-positive neurons in cortex infarct core and cortex infarct penumbra and caudo-putamen of brain regions induced by MCAO. HBCAT treatment after MCAO reduced HSP70- positive neurons in cortex infarct penumbra of brain regions induced by MCAO. Conclusions : These results suggest that HBCAT has a neuroprotective effect against focal cerebral ischemia.