title: A human neural stem cell line provides neuroprotection and improves neurological performance by early intervention of neuroinflammatory system.
abstract: A human neural stem cell line, HB1.F3, demonstrated neuroprotective properties in cerebral ischemia animal models. In this study, we have investigated about the mechanisms of such neuroprotection, mainly focusing on the neuroinflammatory system at an earlier time point of the pathology. Cerebral ischemia model was generated by middle cerebral artery occlusion (MCAO) in adult male Wister rats. HB1.F3 cells were transplanted through jugular vein 6h after MCAO. Forty eight hours after MCAO, transplanted rats showed better neurological performance and decreased TUNEL positive apoptotic cell number in the penumbra. However, haematoxylin and eosin staining and immunostaining showed that, HB1.F3 cells did not affect the necrotic cell death. Twenty four hours after MCAO (18h after HB1.F3 transplantation), infiltrated granulocytes and macrophage/microglia number in the core regions were decreased compared to PBS-treated controls. Immunohistochemical analysis further demonstrated that the transplantation decreased inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 expressing cell number in the core and penumbra, respectively. Double immunofluorescence results revealed that iNOS was mainly expressed in granulocytes and macrophage/microglia in the core region, and COX-2 mainly expressed in neurons, endothelial cells and granulocytes in penumbra. Further analysis showed that although the percentage of iNOS expressing granulocytes and macrophage/microglia was not decreased, COX-2 expressing neurons and vessel number was decreased by the transplantation. In vitro mRNA analysis showed that brain-derived neurotrophic factor (BDNF), basic fibroblast growth factor (βFGF) and bone morphogenic protein (BMP)-4 expression was high in cultured HB1.F3 cells. Thus, our results demonstrated that HB1.F3 cell transplantation provide neuroprotection possibly through the regulation of early inflammatory events in the cerebral ischemia condition.
authors.affiliation: Department of Internal Medicine III, Shimane University School of Medicine, 89-1 Enya Cho, Izumo 693-8501, Japan.
authors.forename: Tatsuzo
authors.initials: T
authors.lastname: Watanabe
authors.affiliation: Department of Laboratory Medicine, Shimane University School of Medicine, 89-1 Enya Cho, Izumo 693-8501, Japan. Electronic address: anagai@med.shimane-u.ac.jp.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Nagai
authors.affiliation: Department of Laboratory Medicine, Shimane University School of Medicine, 89-1 Enya Cho, Izumo 693-8501, Japan.
authors.forename: Abdullah Md
authors.initials: AM
authors.lastname: Sheikh
authors.affiliation: Department of Internal Medicine III, Shimane University School of Medicine, 89-1 Enya Cho, Izumo 693-8501, Japan.
authors.forename: Shingo
authors.initials: S
authors.lastname: Mitaki
authors.affiliation: Department of Internal Medicine III, Shimane University School of Medicine, 89-1 Enya Cho, Izumo 693-8501, Japan.
authors.forename: Kiryo
authors.initials: K
authors.lastname: Wakabayashi
authors.affiliation: Department of Neurology, UBC Hospital, University of British Columbia, Vancouver, Canada; Medical Research Institute, Chung-Ang University College of Medicine, Seoul, Republic of Korea.
authors.forename: Seung U
authors.initials: SU
authors.lastname: Kim
authors.affiliation: Department of Internal Medicine III, Shimane University School of Medicine, 89-1 Enya Cho, Izumo 693-8501, Japan.
authors.forename: Shotai
authors.initials: S
authors.lastname: Kobayashi
authors.affiliation: Department of Internal Medicine III, Shimane University School of Medicine, 89-1 Enya Cho, Izumo 693-8501, Japan.
authors.forename: Shuhei
authors.initials: S
authors.lastname: Yamaguchi
mesh_terms: D000818:Animals; D002545:Brain Ischemia / Q000276:immunology / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D002460:Cell Line; D051546:Cyclooxygenase 2 / Q000378:metabolism; D004195:Disease Models, Animal; D006098:Granulocytes / Q000378:metabolism; D006801:Humans; D020244:Infarction, Middle Cerebral Artery / Q000276:immunology / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D008264:Macrophages / Q000378:metabolism; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D017628:Microglia / Q000378:metabolism / Q000473:pathology; D058953:Neural Stem Cells / Q000166:cytology / Q000637:transplantation*; D015213:Neuroimmunomodulation; D009474:Neurons / Q000378:metabolism / Q000473:pathology; D000066829:Neuroprotection; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
chemical_list: D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat; D051546:Cyclooxygenase 2; C489691:Ptgs2 protein, rat
keywords: HB1.F3 cells; Middle cerebral artery occlusion; Neural stem cell; Neuroinflammation; Neuroprotection