脳血管障害・虚血 Cerebrovascular Disease and Ischemia
O2-9-5-1 ラット海馬歯状回における遅発性神経細胞死を伴った脳梗塞後うつモデル Neurodegeneration in the hippocampal dentate gyrus after transient focal cerebral ischemia in rats: a possible link to post-stroke depression ○笠原二郎1, 内田裕人1, 藤田有貴1, 手塚健太1 ○Jiro Kasahara1, Hiroto Uchida1, Yuki Fujita1, Kenta Tezuka1 徳島大院 ヘルスバイオサイエンス（薬）神経病態解析1 Dept Neurobiol Theapeu, Grad Sch Pharmaceu Sci, Inst Health Biosci, Univ Tokushima, Tokushima, Japan1 Stroke elicits various higher brain dysfunctions in the chronic phase after infarcted. Post-stroke depression is reported to occur in around 40% of patients, although its pathophysiological details are not established with any animal model yet. Using rats, we have been characterizing neurodegenerative changes in distant areas from infarction including substantia nigra and hippocampus after transient focal cerebral ischemia induced by middle cerebral artery-occlusion (MCAO). In this study, we observed delayed neuronal death of the granular cells (GC) in hippocampal dentate gyrus (DG) started to occur around 20 weeks after MCAO, and it was associated with aggravation of anhedonia examined by the sucrose preference test. The increased neuronal death of the matured GC after MCAO was characterized as apoptosis using antibodies for Bax and cleaved caspase-3. On the other hand, proliferation of the neural stem cells in the subgranular zone (SGZ), characterized by antibodies for nestin, glial fibrillary acidic protein (GFAP) and Ki67, was transiently increased immediately after infarction, and then decreased significantly around after 6 weeks of MCAO. These results may suggest underlying mechanisms of the delayed neuronal death in DG. Further more, we examined the effects of chronic antidepressant treatment (imipramine or fluvoxamine) on the behavioral and the neurodegenerative deficits in DG after MCAO. Both aggravation of anhedonia and progression of GC neurodegeneration with reduced proliferation of neural stem cells in SGZ were reduced significantly by chronic treatment with either antidepressant. All of these results suggest it might be an animal model for post-stroke depression associated with neurodegeneration in hippocampal DG. Thus, our findings in this study may provide a novel pathophysiological and therapeutic insight into post-stroke depression after ischemia.	O2-9-5-2 移植ヒト神経幹細胞は中大脳動脈閉塞モデルラットの内在性神経新生を促進し運動機能を回復する Grafted human neural stem cells enhance several steps of endogenous neurogenesis and improve behavioural recovery after middle cerebral artery occlusion in rats ○峯裕1,2,3,4, , 大木宏一2,3,5 ○Yutaka Mine1,2,3,4, Jemal Tatarishvili2,3, Koichi Oki2,3,5, Emanuela Monni2,3, Zaal Kokaia2,3, Olle Lindvall2,3 独立行政法人　国立病院機構　栃木病院　脳神経外科1, ルンド大学病院　神経内科2, ルンド幹細胞センター3, 慶應義塾大学　医学部　生理学教室4, 済生会中央病院　神経内科5 Department of Neurosurgery, Tochigi National Hospital, National Hospital Organization Japan1, Laboratory of Stem cells and Restorative Neurology, Lund University Hospital, SE-221 84 Lund, Sweden2, Lund Stem Cell Center, Lund University Hospital, SE-221 84 Lund, Sweden3, Department of Physiology, Keio University School of Medicine4, Department of Neurology, Saiseikai Central Hospital5 Neural stem/progenitor cells (NSPCs) in subventricular zone (SVZ) produce new striatal neurons during several months after stroke, which may contribute to recovery. Intracerebral grafts of NSPCs can exert beneficial effects after stroke through neuronal replacement, trophic actions, neuroprotection, and modulation of inflammation. Here we have explored whether human fetal striatum-derived NSPC-grafts influence striatal neurogenesis and promote recovery in stroke-damaged brain. T cell-deficient rats were subjected to 1 h middle cerebral artery occlusion (MCAO). Human fetal NSPCs or vehicle were implanted into ipsilateral striatum 48 h after MCAO, animals were assessed behaviorally, and perfused at 6 or 14 weeks. Grafted human NSPCs survived in all rats, and a subpopulation had differentiated to neuroblasts or mature neurons at 6 and 14 weeks. Numbers of proliferating cells in SVZ and new migrating neuroblasts and mature neurons were higher, and numbers of activated microglia/macrophages were lower in the ischemic striatum of NSPC-grafted compared to vehicle-injected group both at 6 and 14 weeks. A fraction of grafted NSPCs projected axons from striatum to globus pallidus. The NSPC-grafted rats showed improved functional recovery in stepping and cylinder tests from 6 and 12 weeks, respectively. Our data show, for the first time, that intrastriatal implants of human fetal NSPCs exert a long-term enhancement of several steps of striatal neurogensis after stroke. The grafts also suppress striatal inflammation and ameliorate neurological deficits. Our findings support the idea that combination of NSPC-transplantation and stimulation of neurogenesis from endogenous NSPCs may become a valuable strategy for functional restoration after stroke.
O2-9-5-3 細胞治療を目標としたマーモセット一過性中大脳動脈閉塞モデルの確立 Development of less invasive transient middle cerebral artery occlusion model on Common Marmoset for cell therapy ○井上賢1,2, 原晃一3, 疋島啓吾4, 小牧裕司4, 伊藤豊志雄4, 岩田祐士5, 武見充晃6, 牛場潤一7, 塚田秀夫8, 岡野ジェイムス洋尚9, 岡野栄之2 ○Satoshi Inoue1,2, Kohich Hara3, Keigo Hikishima4, Yuji Komaki4, Toshio Itoh4, Hiroshi Iwata5, Mitsuaki Takemi6, Junichi Ushiba7, Hideo Tsukada8, James Hirotaka Okano9, Hideyuki Okano2 慶應大院・医・脳神経外科1, 慶應義塾大学　生理学教室2, 日野市立病院　脳神経外科3, 実験動物中央研究所4, 島津製作所5, 慶應義塾大学　理工学研究科6, 慶應義塾大学　理工学部生命情報学科7, 浜松ホトニクス　中央研究所　PETセンター8, 東京慈恵会医科大学　再生医学研究9 Dept Neurosurgery, Univ of Keio, Tokyo1, Dept Physiol, Univ of Keio, Tokyo2, Hino Municipal Hospital, Tokyo3, Central Institute for Experimental Animals, Kawasaki4, Shimadzu Corporation, Kyoto5, Science and Technology, Univ of Keio, Yokohama6, Science and Technology, Univ of Keio, Yokohama7, Hamamatsu Photonics K.K. PET Center, Hamamatsu8, Regenerative medicine ,Univ of Jikei, Tokyo9 [Objective] We have developed a less invasive transient middle cerebral artery occlusion (MCAO) model on primates, common marmosets, without invasive procedure including craniotomy. Behavioral assessments (motor paresis, sensory disturbance and visual field defect) and MRI were performed with time postoperatively. In addition, Positron emission tomography (PET) was examined (performed) for assessments of ischemic brain. [Method] Female laboratory-bred common marmosets were used. Under anesthesia, wire thread was inserted into right middle cerebral artery (MCA) via carotid artery. Intraoperative MRI or X-ray was done to confirm the MCA occlusion. We used 3 hours occlusion. Behavioral assessments, MRI and PET were performed postoperatively. In PET study, we used BCPP-EF, which is ligand to the complex1 of mitochondria. [Result] At postoperative day 7, MRI T2 weighted image showed brain infarction on right hemisphere including basal ganglia as high intensity lesion. However, after 28 days, the high intensity area was reduced dramatically. In PET, BCPP-EF showed the comparable result. Behavioral tests exhibited neurological deficits with left hemiparesis deteriorating for 4 days after surgery. After that, neurological function improved gradually. Six weeks after surgery, the behavior did not recover completely. Remained motor paresis such as ham hand could be observed by food retrieval test. The immunohistochemical study also showed defect of neural cells on the lesion. [Conclusion] We have developed less invasive marmoset MCAO model. MRI, PET and behavioral tests showed reasonable brain infarction as well as human. The model is considered to be useful for examination of neurogensis and therapeutic efficacy on brain infarction.	O2-9-5-4 A new model of lacunar stroke in the non-human primate ○Sandra Puentes1, Takanobu Kaido2, Takashi Hanakawa3, Taisuke Otsuki2, Kazuhiko Seki1 Dept Neurophysiol, Nat Instit of Neurosci, Tokyo Japan.1, Dept Neurosurg, Nat Center of Neurol and Psych, Tokyo Japan.2, Dept Mol Imag, Nat Center of Neurol and Psych, Tokyo Japan.3 Motor dysfunction including hemiparesis, incoordination and spasticity are commonly generated after stroke. Nevertheless standard rehabilitation interventions typically correlate with modest rather than marked improvements in motor function. To understand in detail the motor disturbances after stroke and establishing effective interventions, animal stroke models that can induce isolated motor impairment relevant to human patients are required. As middle cerebral artery occlusion (MCAO) is among the most common stroke presentations, several animal models have been developed to reproduce this condition. Although experimental MCAO can induce motor impairment, the infarct compromises several brain structures that impede isolated motor evaluation. The goal of this study was to develop an ischemia model in marmosets by occluding the anterior choroidal artery (AChA) to impair the internal capsule (IC). Initially, the vascular distribution of the marmoset brain was examined by intra-arterial colored latex perfusion finding a high similarity between the marmoset and human vascular anatomy. A unique AChA was found in 10 cases, and four cases showed duplicated or triplicated vessels converging in a unique artery. After confirm its anatomical distribution, animals were operated to electrocoagulate the AChA and euthanized few hours after the procedure; chronic experiments also were performed and compared with sham-operated animals 11 days after surgery. Animals were evaluated by using MRI (day 4) and histopathology (day 11). For chronic experiments, AChA occluded animals showed an infarction localized mainly in the IC with minimal damage to the basal ganglia. No damage was found in the IC of the sham-operated animals. We developed a new technique to induce a selective infarction of the posterior limb of the IC. This model will be useful for the development of new strategies for patients with corticospinal tract injury after stroke.

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