• 대한본초학회지
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• 제24권1호
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• pp.179-189
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• 2009

Objectives : Coptidis Rhizoma (Coptis japonica MAKINO; CR) is a well known crude drug as antimicrobial, antibacterial, anti-inflammatory, antioxidant activity. However, there is no study of the effect of CR on brain infarction and it's mechanism. The aim of this study was to investigate the effects on ischemic stroke induced by photothrombotic infarction by evaluating the functional & neuronal recovery after brain infarction. Materials & Methods : Male Sprague-Dawley rats (250-300 g) were induced photothrombotic brain infarction on sensorimotor cortex, and brain infarction volume by image J software (NIH, USA) after Nissl stain, also single pellet reaching task as a functional motor recovery were observed. After orally pretreated by CR (500 mg/kg) or normal saline as a sham control before 7 days from the time of photothrombotic infarction, rats were sacrificed. After then we analysed anti-inflammatory cytokines (TNF-$\alpha$, IL-6, IL-1$\beta$), by RT-PCR and ELISA method, and immunohistochemistry (GFAP, connexin-43) as a marker of neural plasticity. Results : CR (100, 250, 500 mg/kg) decreased the infarction volume dose-dependently, however the effect of 500mg/kg of CR (CR 500) showed the best (P=0.051). Also, CR 500 decreased the infarction volume time-dependently, the most effective time was 3-7 days after stroke. Photothrombosis increased inflammatory cytokines after infarction, CR 500 suppressed significantly mRNA expression of IL-1$\beta$, IL-6 and TNF-$\alpha$. In serum, CR 500 decreased the amount of IL-1$\beta$, 12h, 24h and 48h respectively (p < 0.05), also decreased that of IL-6 and TNF-$\alpha$, 12h respectively (p < 0.05) after infarction. The more astrocytes were observed and neural plasticity was facilitated in the rat brain of CR 500 than that of sham control in immunohistochemistry. Conclusions : This results suggest that CR decrease infarction volume and improve functional motor recovery in acute stage in photothrombotic ischemic infarction model in the mechanism of anti-inflammation and promoting neural plasticity.

• 동의생리병리학회지
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• 제23권3호
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• pp.645-661
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• 2009

The water extract of Dohongsamul-Tang(DHSMT) has been traditionally used to stroke and brain injuries in Oriental Medicine. The present study was designed to investigate the effects of DHSMT on the gene expression profile of cerebral infarction by cDNA microarray in photothrombotic ischemia mouse model. Photothrombotic ischemia was induced in stereotactically held male BALB/c mice using rose bengal and cold light. MRI was performed 24 hours after inducing photothrombosis using 1.5 T MRI and 47 mm surface coil to obtain T2-weighted, and contrast-enhanced images. After MRI test, animal was sacrificed and the brain sections were stained for hematoxylin and eosin and immunohistochemistry. MRI and histological analysis revealed that lesion of thrombotic ischemia was well induced in the cortex with the evidence of biological courses of infarction. The target area of thrombotic infarction was 1 mm anterior to bregma and 3 mm lateral to midline with 2 mm in diameter, which were decreased by administration of DHSMT. To assess gene expression pattern of cerebral infarction, mRNA was isolated and reacted with microarray chip(Agilant's DNA Microarray 44K). Scatter and MA plot analysis were performed to clustering of each functional genes. M value [M=log2(R/G), A={log2(R ${\times}$ G)}/2] was between -0.5 and +0.5 with 40% difference. After pretreatment with DHSMT, the expression levels of mRNA of many genes involved in various signaling pathway such as apoptosis, cell cycle, cell proliferation, response to oxidative stress, immune response, angiogenesis, and inflammatory cytokine were markedly inhibited in photothrombotic ischemia lesion compared to the control group. These results suggest that DHSMT prevent ischemic death of brain on photothrombotic ischemia model of mice through modulation of gene expression at the transcriptional level.

• Journal of Korean Neurosurgical Society
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• 제54권6호
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• pp.467-476
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• 2013

Objective : This study aimed to evaluate the hypotheses that administration routes [intra-arterial (IA) vs. intravenous (IV)] affect the early stage migration of transplanted human bone marrow-derived mesenchymal stem cells (hBM-MSCs) in acute brain infarction. Methods : Male Sprague-Dawley rats (n=40) were subjected to photothrombotic infarction. Three days after photothrombotic infarction, rats were randomly allocated to one of four experimental groups [IA group : n=12, IV group : n=12, superparamagnetic iron oxide (SPIO) group : n=8, control group : n=8]. All groups were subdivided into 1, 6, 24, and 48 hours groups according to time point of sacrifice. Magnetic resonance imaging (MRI) consisting of T2 weighted image (T2WI), $T2^*$ weighted image ($T2^*WI$), susceptibility weighted image (SWI), and diffusion weighted image of rat brain were obtained prior to and at 1, 6, 24, and 48 hours post-implantation. After final MRI, rats were sacrificed and grafted cells were analyzed in brain and lung specimen using Prussian blue and immunohistochemical staining. Results : Grafted cells appeared as dark signal intensity regions at the peri-lesional zone. In IA group, dark signals in peri-lesional zone were more prominent compared with IV group. SWI showed largest dark signal followed by $T2^*WI$ and T2WI in both IA and IV groups. On Prussian blue staining, IA administration showed substantially increased migration and a large number of transplanted hBM-MSCs in the target brain than IV administration. The Prussian blue-positive cells were not detected in SPIO and control groups. Conclusion : In a rat photothrombotic model of ischemic stroke, selective IA administration of human mesenchymal stem cells is more effective than IV administration. MRI and histological analyses revealed the time course of cell migration, and the numbers and distribution of hBM-MSCs delivered into the brain.

• 동의생리병리학회지
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• 제26권6호
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• pp.894-901
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• 2012

The effect of Sopung-tang(SPT) and trans-cranial direct current stimulation(tDCS) was investigated in photothrombotic brain infarction(PTI) rats. Sprague-Dawley 80 rats, were divided into four groups. group I was experiental control group(n=20), group II was PTI induced and oral administration of SPT(n=20), group III was PTI induced and tDCS administration(n=20) and group IV was PTI induced and SPT and tDCS administration for 28 days on stroke rats(n=20). Analysis the neurological function test, 25 point behavior functional score test, and immunohistochemistric finding of GDNF expression, and electron microscopy assessment In motor behavior test, the outcome of group IV was significantly difference than the other group, and In immunohistochemistric finding, group II, III, IV were increase GDNF expression on 28 days, In electron microscopy finding, the all groups were degenerated of cell organelles, and synaptic plasticity were improvement of group II, III, IV(especially group IV) These results suggest that, 28days application of SPT and tDCS was the motor function and histopathologic, micro-morphological improvement of motor function recovery and positive influence on synaptic plasticity.

• 대한의생명과학회지
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• 제29권4호
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• pp.263-273
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• 2023

This study aimed to investigate effect of scalp acupuncture (SA) and repetitive transcranial magnetic stimulation (rTMS) intervention on neuromotor function in photothrombotic cerebral infarction (PCI) rat model. Sixty male SD rats were used. PCI was induced on M1 cortex of right frontal lobe. SA was performed at the Qianding (GV21), Xuanli (GB6) acupoints of ipsilesional M1. Low-frequency rTMS was delivered to contralesional M1. All rats were randomly divided into 4 groups: group A, normal (n, 15); group B, PCI without any stimulation intervention (n, 15); group C, PCI with SA (n, 15); group D, PCI with rTMS (n, 15). Rota-rod test and Ladder rung walking test (LWT) were done weekly for 8 weeks after PCI. SA or rTMS was started from post-PCI 4^th day as protocol for 8 weeks. H/E stain and IHC were done. Western blot and qRT-PCR study were performed for MAP2 and BDNF from ipsilesional M1 peri-infarction tissue. Brain MRI study was conducted to quantify the volume of cerebral infarction. As a result, left forelimb and hindlimb function significantly improved more in group C and D than control group, with expressed more BDNF and MAP2. And brain MRI showed focal infarction of right M1 after PCI, and infarction volume progressively decreased in group C and D than group B from post-PCI 5^th to 8^th week. SA or rTMS was more effective than no intervention group on neuromotor function of PCI rat model. The functional recovery was associated with stimulation intervention-related neurogenesis.

• 한국콘텐츠학회논문지
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• 제10권8호
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• pp.249-256
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• 2010

사람 중간엽 줄기세포(hMSCs)를 초상자성산화철(SPIO)로 표지하여, 체외에서 세포 농도에 따른 신호의 변화를 알아보고, 흰쥐 광 혈전 뇌경색 모델에 이식하여 자기공명영상 기법 중, T2강조영상과 $T2^*$강조영상 그리고 자화강조영상의 줄기세포 검출능을 분석하고, 병리 소견과 비교하고자 하였다. 체외실험은 SPIO(Feridex I.V.$^{(R)}$)로 표지한 줄기세포 $1.56{\times}10^4$, $3.13{\times}10^4$, $6.25{\times}10^4$, $1.25{\times}10^5$, $2.5{\times}10^5$, $5{\times}10^5$ 개/$m{\ell}$, SPIO로 표지 하지 않은 $5{\times}10^5$ 개/$m{\ell}$의 세포를 Control로 준비하여 $0.5m{\ell}$튜브에 담아 T2WI, $T2^*WI$, SWI sequence로 자기공명영상을 얻었다. 체내실험은 8마리의 흰쥐를 대상으로 정수리점(bregma)에서 우측으로 2.5mm, 뒤쪽으로 2.5mm 부위에 광 혈전 뇌경색을 만들고, SPIO-hMSC $2.5{\times}10^5$ 개/$m{\ell}$, $1m{\ell}$을 꼬리 정맥에 주입하여 실험하였다. 자기공명영상은 주입 전과 주입 후 1, 3, 7 그리고, 14일째 T2WI, $T2^*$WI, SWI 영상을 얻고 정량적으로 평가하였다. 자기공명영상검사 후 흰쥐는 조직검사를 위해 희생시켰다. 체외실험 결과 CNR은 SWI, $T2^*WI$, T2WI 순으로의 높게 나왔고, 세포 농도에 따라서는 $2.5{\times}10^5$ 개/$m{\ell}$의 농도에서 가장 높게 나왔다. 체내실험에서 normal, infarction, SPIO 상호간의 CNR을 분석해본 결과 normal-infarction의 CNR은 T2WI가 가장 높게 나왔고, normal-SPIO 그리고 infarction-SPIO에서 CNR은 SWI, $T2^*WI$, T2WI순으로 나왔다. 따라서 흰쥐를 이용한 실험에서 T2WI sequence는 infarction을 잘 표현해 주고, SWI는 SPIO로 표지한 줄기세포를 normal과 infarction으로 부터 잘 구별해 주었다. 현재 임상에서 SWI는 출혈, 석회화 병변등을 쉽게 찾아내는 sequence로 사용 되고 있지만 향후에 줄기세포 치료가 인간의 질병을 치료하는 주요방안으로 자리 잡을 때에도 이식된 줄기 세포를 관찰 할 수 있는 좋은 도구가 될 것으로 기대 된다.

• Journal of Korean Neurosurgical Society
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• 제40권3호
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• pp.180-185
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• 2006

Objective : Middle cerebral artery occlusion[MCAO] has widely been used to produce ischemic brain lesions. The lesions induced by MCAO tend to be variable in size because of the variance in the collateral blood supply found in the mouse brain. To establish a less invasive and reproducible focal ischemia model in mice, we modified the technique used for rat photo thrombosis model. Methods : Male C57BL/6 mice were subjected to focal cerebral ischemia by photothrombosis of cortical microvessels. Cerebral infarction was produced by intraperitoneal injection of Rose Bengal, a photosensitive dye and by focal illumination through the skull. Motor impairment was assessed by the accelerating rotarod and staircase tests. The brain was perfusion-fixed for histological determination of infarct volume four weeks after stroke. Results : The lesion was located in the frontal and parietal cortex and the underlying white matter was partly affected. A relatively constant infarct volume was achieved one month after photothrombosis. The presence of the photothrombotic lesion was associated with severe impairment of the motor performance measured by the rotarod and staircase tests. Conclusion : Photothrombotic infarction in mice is highly reproducible in size and location. This procedure can provide a simple method to produce cerebral infarction in a unilateral motor cortex lesion. In addition, it can provide a suitable model for study of potential neuroprotective and therapeutic agents in human stroke.

• 대한디지털의료영상학회논문지
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• 제13권2호
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• pp.55-58
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• 2011

It is important to study using experimental animals for research about stroke. Magnetic Resonance Imaging(MRI) is avaluable diagnosticmethods for stroke diagnosis. The purpose of this research is to know the Magnetic Resonance Imaging(MRI) and histopathological characteristics findings after induction of photothrombotic cerebral infarction in rat brain. Male Sprague-Dawley rats were anesthetized, Rose Bengal dye(20 mg/kg) was intravenously injected. The right sensonrimotor cortex of rat brain was exposed to cold light of 7 mm diameter at a position of 1 mm anterior and 3.5 mm lateral to bregma for 20 min. The post-infarction effects were monitored by T1 weighted and T2 weighted images of brain MRI. Histopathological changes were observed after Hematoxylin & Eosin staining. The lesion appeared clearly high signal intensity area on T2 weighted images(the major axis $7.04{\pm}0.11$ mm, the minor axis $3.08{\pm}0.04$ mm) and also H&E staining was same result. In conclusion, MRI was avaluable diagnostic methods for diagnosis and serial changes of stroke.

• The Korean Journal of Physiology and Pharmacology
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• 제22권1호
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• pp.101-110
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• 2018

In this study, we aimed to investigate the neuroprotective effects of caffeic acid phenethyl ester (CAPE), an active component of propolis purified from honeybee hives, on photothrombotic cortical ischemic injury in mice. Permanent focal ischemia was achieved in the medial frontal and somatosensory cortices of anesthetized male C57BL/6 mice by irradiation of the skull with cold light laser in combination with systemic administration of rose bengal. The animals were treated with CAPE (0.5-5 mg/kg, i.p.) twice 1 and 6 h after ischemic insult. CAPE significantly reduced the infarct size as well as the expression of tumor necrosis $factor-{\alpha}$, hypoxiainducible $factor-1{\alpha}$ monocyte chemoattractant protein-1, $interleukin-1{\alpha}$, and indoleamine 2,3-dioxygenase in the cerebral cortex ipsilateral to the photothrombosis. Moreover, it induced an increase in heme oxygenase-1 immunoreactivity and interleukin-10 expression. These results suggest that CAPE exerts a remarkable neuroprotective effect on ischemic brain injury via its anti-inflammatory properties, thereby providing a benefit to the therapy of cerebral infarction.