外傷 Injury
P3-2-228 Ndrg2は脳損傷後のアストロサイト活性化を促進する Ndrg2 promotes astroglial activation after brain injury ○宝田美佳1, 毛塚大1, 武市敏明2, 北尾康子1, 堀修1 ○Mika Takarada-Iemata1, Dai Kezuka1, Toshiaki Takeichi2, Yasuko Kitao1, Osamu Hori1 金沢大院・医・神経分子標的学1, 金沢医大・医・法医学2 Dept Neuroanat, Kanazawa Univ, Kanazawa, Japan1, Dept Legal Med, Kanazawa Med Univ, Kanazawa, Japan2 N-myc downstream-regulated gene 2 (Ndrg2) is a member of recently identified differentiation-related genes, and is predominantly expressed in astrocytes in the central nervous system. We recently reported that Ndrg2 is upregulated in the mouse model of Parkinson's disease, and may regulate astroglial activation during neurodegeneration (Takeichi et al., 2011). This study was aimed to elucidate the possible function and the importance of Ndrg2 in astroglial activation using stab wound injury model. We established Ndrg2 global knockout (KO) mouse by crossing Ndrg2-floxed mice with CAG-Cre mice. The absence of Ndrg2 expression in Ndrg2 KO mice was confirmed by RT-PCR and Western blot analysis. In wild-type (WT) mice, enhanced level of expression of Ndrg2 was observed in GFAP-positive reactive astrocytes surrounding the wounded area from 1 to 7 days after injury. In contrast, in Ndrg2 KO mice, expressions of GFAP and other astrogliosis markers such as Lcn2, Serpina3n, and phosphorylated STAT3 were at lower levels compared to those in WT mice. Consistent with these results, immunohistochemical analysis revealed that glial scar formation was attenuated in Ndrg2 KO mice after injury. Furthermore, qPCR and immunohistochemical analysis revealed that induction of astroglial activators such as IL-6 were impaired in Ndrg2 KO mice at 1 day after injury. These results suggest that Ndrg2 may contribute to astroglial activation through the induction of astroglial activators.	P3-2-229 脊髄損傷マウスに対し改善効果を持つ漢方生薬の研究 Investigation of crude drugs effective for spinal cord injured mice ○田辺紀生1, 久保山友晴1, 宮永賢2, 数馬恒平2, 紺野勝弘2, 東田千尋1 ○Norio Tanabe1, Tomoharu Kuboyama1, Satoshi Miyanaga2, Kouhei Kazuma2, Katsuhiro Konno2, Chihiro Tohda1 富山大・和漢研・神経機能学1, 富山大・和漢研・和漢薬製剤開発2 Div of Neuromedical Science, Inst of Natural Med, Univ of Toyama1, Div of Kampo-Pharmaceutics, Inst of Natural Med, Univ of Toyama2 After spinal cord injury (SCI), astrocytes are increased and secrete chondroitin sulfate proteoglycan (CSPG) in the injured region. Since the CSPG inhibits axonal extension, broken neuronal network is hardly reconstructed, and motor dysfunction is not restored. Therefore, axonal extension in the inhibitory environment is one of essential factors for recovery from motor dysfunction. In this study, we aimed to identify drugs that were able to induce axonal extension on the inhibitory CPSG substrate and improved motor function in SCI mice. Water extracts of crude drugs used in Kampo medicine (110 kinds) were screened for axonal extension activity. The water extract of each crude drug (1 or 10 μg/ml) or vehicle solution was added to the cortical neurons (ddY mice, E14) that were cultured on the CSPG. Four days after, axonal lengths were evaluated by immunocytochemistry for pNF-H. Although axonal extension was inhibited on the CSPG substrate, water extracts of crude drug S, crude drug A and crude drug F strongly induced axonal extension even on CSPG. Next, we investigated effects of those 3 kinds of extracts using SCI mice (ddY, female, 8 weeks old) suffered from contusion injury. Crude drugs S, A and F (300 mg/kg/day) or vehicle solution was orally administrated to SCI mice 1 h after the injury and once a day for following 30 days. Crude drugs S and F significantly recovered the motor function of hindlimbs in SCI mice, evaluated by Basso Mouse Scale and Body Support Scale, but crude drug A showed no effect. In this study, we firstly demonstrated improvement effects of crude drugs S and F on SCI mice. For aiming to develop new therapeutic medicines for SCI, we are now identifying active constituents in S and F and clarifying underlying mechanisms. In addition, the possibility of application of those crude drugs as Kampo formula is also studied.
P3-2-230 瀰漫性軸索損傷後の認知機能障害および精神症状 Cognitive impairments and psychiatric symptoms in patients with diffuse axonal injury ○生方志浦1,2, 上田敬太1, 久保田学1, 吉住美保1, 種村留美2, 澤本伸克3, 福山秀直3 ○Shiho Ubukata1,2, Keita Ueda1, Manabu Kubota1, Miho Yoshizumi1, Rumi Tanemura2, Nobukatsu Sawamoto3, Hidenao Fukuyama3 京都大院・医・精神医学1, 神戸大院・保・リハ科学2, 京都大院・医・高次脳機能センター3 Dept Psychiatry, Univ of Kyoto, Kyoto1, Dept Helthscience, Univ of Kobe, Kobe2, Human Brain Research Center, Univ of Kyoto, Kyoto3 Focal brain injury and diffuse axonal injury (DAI) are two typical types of injury pattern in traumatic brain injury. It is well known that patients with frontal lobe injury show psychiatric symptoms and impairments in cognition. On the other hand, there are few studies that investigate the neurocognitive and psychiatric sequelae after DAI. In this study, the neuropsychological sequelae after DAI were examined in comparison with those of frontal focal lesion. Nine patients with DAI (mean age = 32.1, SD = 10.2 years), 11 patient with focal lesion (mean age = 44.8, SD = 15.7), and 12 healthy control subjects underwent magnetic resonance imaging (MRI). All subjects completed neuropsychological assessments and measures for psychiatric symptoms. We performed voxel-based morphometry (VBM) to investigate regional brain alterations. There are significant differences in basic cognition between those with focal lesion and those with DAI. But as to social cognition, both groups were impaired to the same degree. Patients with DAI showed some psychiatric symptoms such as apathy, depression and compulsive symptom. And to our surprise, significant difference was found only in apathy scale and focal lesion patients showed severer apathetic symptom. We will be discussing these results, including the relationship between neuropsychiatric sequelae and brain morphology.	P3-2-231 エダラボンは頭部外傷による軸索損傷と酸化ストレスを阻止し、記憶障害を改善する Attenuation of axonal injury and oxidative stress by edaravone protects against cognitive impairments after traumatic brain injury ○東洋一郎1, 大田学1, 八幡俊男1, 北原正大1, 藤本康倫1, 清水恵司1 ○Youichirou Higashi1, Manabu Ota1, Toshio Yawata1, Masahiro Kitahara1, Yasunori Fujimoto1, Keiji Shimizu1 高知大学 医学部 脳神経外科1 Dept. Neurosurg, Kochi Univ, Kochi1 Traumatic axonal injury (TAI), a feature of traumatic brain injury (TBI), progressively evolves over hours through impaired axonal transport and may be a major contributor to cognitive dysfunction. In spite of various studies suggesting that pharmacologic or physiologic interventions might reduce TAI, clinical neuroprotective treatments are still unavailable. Edaravone, a free radical scavenger, has been shown to exert neuroprotective effects in animal models of several brain disorders. In this study, to evaluate whether edaravone suppresses TAI following TBI, mice were subjected to weight drop injury and had either edaravone (3.0 mg/kg) or saline administered intravenously immediately after the impact. Axonal injury and oxidative stress were assessed using immunohistochemistry with antibodies against amyloid precursor protein, a marker of impaired axonal transport, and with 8-hydroxy-2-deoxyguanosine, a marker of oxidative DNA damage. Edaravone significantly suppressed axonal injury and oxidative stress in the cortex, corpus callosum, and hippocampus 24h after the injury. The neuroprotective effects of edaravone were observed in mice receiving 1.0, 3.0, or 10.0 mg/kg of edaravone immediately after impact, but not after 0.3 mg/kg of edaravone. With treatment 1h after the impact, axonal injury was also significantly suppressed and this therapeutic effect persisted up to 6h after the impact. Furthermore, behavioral tests performed 9 days after injury showed memory deficits in saline-treated traumatized mice, which were not evident in the edaravone-treated group. These results suggested that edaravone protects against memory deficits following TBI and that this protection is mediated by suppression of TAI and oxidative stress.
P3-2-232 Secretion of ATP from Schwann cells through lysosomal exocytosis during Wallerian degeneration ○Younho Shin1, Seo Jin Lee1, Byung Sun Park1, Youngbuhm Huh1, Junyang Jung1 Kyung Hee University1 The present study demonstrates that adenosine triphosphate (ATP) is released from Schwann cells through lysosomal exocytosis during Wallerian degeneration and in response to stimulation. In primary Schwann cell cultures, ATP was stored in lysosomal vesicles. ATP could then induce Ca(2+)-dependent lysosomal exocytosis. Among three stimulants of lysosomal exocytosis (glutamate, NH(4)Cl and zymosan), only NH(4)Cl was sufficient to induce ATP release from ex vivo sciatic nerve explants at 3days in vitro. Lysosomal exocytosis inhibitors (metformin, chlorpromazine and vacuolin-1) reversed the effect of NH(4)Cl-enhanced ATP release, replicating the state of explants treated with NH(4)Cl in the absence of lysosomal exocytosis inhibitors. Furthermore, we observed ATP release through lysosomal exocytosis during Wallerian degeneration in sciatic explant cultures using the recently identified vesicular nucleotide transporter (VNUT). From these experiments, we conclude that the exocytosis of lysosomes in Schwann cells during Wallerian degeneration is Ca(2+)-dependent, and that it induces ATP release from Schwann cells.

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