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化合物CHO3对LPS激活小胶质细胞BV2炎症的影响

化合物CHO3对LPS激活小胶质细胞BV2炎症的影响

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者：宋修云胡金凤陈乃宏中国医学科学院药物研究所

目的化合物CH03是我所自主研究开发的新型小分子化合物,本实验室前期研究发现其对多种诱导剂所致神经细胞损伤均具有显著改善作用。本研究旨在观察化合物CH03对脂多糖（LPS）所致小鼠小胶质细胞瘤细胞BV2激活的抑制作用。方法 100 ng/mL... 详细信息

目的化合物CH03是我所自主研究开发的新型小分子化合物,本实验室前期研究发现其对多种诱导剂所致神经细胞损伤均具有显著改善作用。本研究旨在观察化合物CH03对脂多糖（LPS）所致小鼠小胶质细胞瘤细胞BV2激活的抑制作用。方法 100 ng/mL LPS作用BV2细胞构建神经炎症细胞模型,采用可见光观察、MTT法、ELISA、Gress法、DCF-DA荧光探针、Western Blot、免疫荧光检测化合物CH03（0.1,1,10μmol/L）对BV2细胞激活的影响。结果形态学观察结果表明,LPS模型组细胞呈阿米巴状,1、10μmol/L CH03组,细胞折光度良好,阿米巴状细胞数明显减少。MTT结果表明,与空白对照及LPS组比较,CH03各剂量对细胞存活率均无显著影响,说明CH03对静息及活化状态下的BV-2细胞存活率均无显著影响。ELISA结果显示,LPS组的细胞培养上清中TNF-α、IL-1β含量分别为2059±33pg/mL、169±5pg/mL（空白对照组为54±14pg/mL、58±11pg/mL）,而0.1、1、10μmol/L CH03组TNF-α含量分别为2065±12 pg/mL、1954±28 pg/mL、1506±35 pg/mL,IL-1β含量分别为172±6 pg/mL、130±6 pg/mL、88±8 pg/mL,即1、10μmol/LCH03均能显著抑制炎症因子TNFα、IL-1β的释放。Gress实验结果表明,LPS组细胞培养上清中NO含量为100.0%±0.5%（空白对照组28.9%±0.5%）,0.1、1、10μmol/L CH03组NO含量分别为95.7%±1.6%、89.4%±1.7%、79.1%±4.2%,即CH03可显著抑制LPS诱导的BV2细胞生成NO。DCF-DA荧光探针检测胞内ROS结果显示,1、10μmol/L CH03组细胞绿色荧光强度较LPS组显著降低。Western Blot、免疫细胞荧光结果显示,CH03可剂量依赖性抑制LPS诱导的NF-κB/p65入核,抑制JNK、p38的磷酸化,抑制iNOS、COX-2胞内表达。即CH03可通过抑制NF-κB入核、MAPK通路的活化,抑制炎症蛋白iNOS、COX-2表达,进而抑制炎症因子ROS、NO、TNF-α、IL-1β的生成。结论化合物CH03对LPS所致BV2细胞炎症具有明显保护作用,提示CH03可能具有抗神经炎症的作用,对于对抗神经退行性疾病的研究可能具有重要意义。

关键词：化合物CH03 脂多糖 BV2细胞神经炎症

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HMGB1的两种亚型具有不同调节功能

HMGB1的两种亚型具有不同调节功能

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者：王勇军董莹莹宦有娟丁斐顾晓松南通大学江苏省神经再生重点实验室

目的近年来研究发现,免疫细胞可以被动或主动释放高迁移率族蛋白HMGB1,进而调节炎症反应。同时也有研究认为,HMGB1促进神经元细胞黏附和轴突生长,参与神经系统的发育和成熟。然而这两种矛盾的生物学功能如何在同一分子中得到统一,一直... 详细信息

目的近年来研究发现,免疫细胞可以被动或主动释放高迁移率族蛋白HMGB1,进而调节炎症反应。同时也有研究认为,HMGB1促进神经元细胞黏附和轴突生长,参与神经系统的发育和成熟。然而这两种矛盾的生物学功能如何在同一分子中得到统一,一直令人疑惑。我们以多疣壁虎为实验模型,克隆了HMGB1基因的两个亚型HMGB1a和HMGB1b,本实验拟对这两个不同亚型基因进行功能比较研究。方法壁虎腹腔注射LPS以及断尾再生研究HMGB1a和HMGB1b的表达;以RAW264.7巨噬细胞株和神经母细胞瘤进行体外表达分析以及功能位点的突变分析。结果（1）同源比对发现,HMGB1a和HMGB1b的主要功能域仅有一个氨基酸不同。壁虎断尾再生模型研究发现,HMGB1a和HMGB1b主要定位于脊髓的灰质,断尾损伤后HMGB1b的表达呈明显上调,而HMGB1a的表达丰度在断尾损伤后没有显著性差异;（2）LPS腹腔注射模型显示,在主要免疫调节器官脾脏以及肾和肝中,HMGB1a的表达随着处理时间的延长,呈逐渐上升趋势,而HMGB1b在各组织中表达变化无明显差异;（3）体外实验结果显示,在RAW264.7巨噬细胞株中过表达HMGB1a后,免疫因子TNF-α、IL1-a表达量显著上调,而过表达HMGB1b后免疫因子TNF、IL1a表达量不发生明显变化;对活性位点的氨基酸互换突变后发现,过表达HMGB1a其效应与HMGB1b类似,而过表达HMGB1b后效应与HMGB1a类似;（4）在神经元细胞中的研究发现,过表达HMGB1b后,神经元细胞Bcl-2/Bax显著升高,而过表达HMGB1a则没有显著变化,活性位点的氨基酸进行互换突变后过表达发现,HMGB1a和HMGB1b的功能也发生互换。研究表明,HMGB1a和HMGB1b主要通过TLR2和TLR4受体起作用。结果提示,HMGB1a和HMGB1b分别与免疫调节和神经元抗凋亡有关,哺乳动物中的HMGB1促进神经再生以及介导免疫调节,可能由HMGB1的不同亚型调节。

关键词：壁虎 HMGB1 亚型凋亡炎症

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The neurovascular imaging in cerebral ischemia by laser speckle contrast analysis

The neurovascular imaging in cerebral ischemia by laser spec...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Shan-Bao TONG~*,Peng MIAO,Min-Heng LI,Hong-Yang LU,Qi LIU,Yao LI School of Biomedical Engineering,Shanghai Jiao Tong University,Shanghai 2000240,China

Objective Laser speckle contrast analysis has been used in imaging the cerebral vasculature and cerebral blood flow(CBF).However,in practice,the estimation of the CBF may be biased by the ambient background field, dark current and other anomalies in the circuits of CCD camera,non-uniform distributed laser illumination,cortical curvature,breathing/heart beating of the subjects,and the nonstationary change of CBF under functional stimulation, and the motion artifacts if applied to the behavioring *** In order to obtain a stable and high spatiotemporal resolution CBF imaging in the study of the ischemic brain injury,we developed several techniques to improve the current techniques of laser speckle contrast analysis,including(1) enhanced laser speckle contrast analysis (eLASCA);(2) model Based LSI reconstruction;(3) registered laser speckle contrast analysis(rLASCA);(4) Random process estimator for laser speckle *** finally we integrate these techniques and designed a miniature headstage imaging system which can be used to obtain the CBF imaging when the animal(***) is moving freely. Results By using the above techniques,high resolution CBF information can be obtained in real time for either anesthetized or freely moving *** We developed high-resolution optical imaging technique for CBF monitoring,which can be used for real time monitoring both structural and functional neurovascular changing in ischemic brain injury.

关键词： neurovascular imaging cerebral blood flow laser speckle imaging laser speckle contrast analysis

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A practical model of bone inflammation induced pain by intra-tibial CFA injection in rats

A practical model of bone inflammation induced pain by intra...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Chang-Jiang YANG,Shan HU,Qian LI,Ping HAN,Gen-Cheng WU,Yan-Qing WANG,Qi-Liang MAOYING~* Department of Integrative Medicine and Neurobiology,Shanghai Medical College,Institute of Acupuncture Research, Institutes of Brain Science,State Key Laboratory of Medical Neurobiology,Fudan University,Shanghai,China

Background In prior studies,models of inflammatory pain have been produced through injecting complete Freund's adjuvant(CFA) or capsaicin directly into either the deep somatic tissue or the plantar portion of animal hind *** contrast,bone cancer pain is simulated in animal by injecting tumor cells into the cavity of the femur or *** has been reported that the same stimulus to various tissues types might result in differing patterns of pain response due to differences in afferent *** The present study was designed to establish a rat model of inflammatory bone pain to better understand bone cancer *** Our rat model of inflammatory bone pain was produced through intra-tibial injection of *** in body weight,bone histology,mechanical allodynia and thermal hyperalgesia were *** effect of Celecoxib on inflammatory bone pain was also *** Significant inflammation was seen in the bone marrow two days after intra-tibial CFA injection, including nuclear condensation and fragmentation,massive neutrophilic granulocytes,and prominent fibrinous *** days after injection,marked fibrosis of the bone was detected by histological *** unilateral CFA injection,behavioral studies showed mechanical allodynia to von Frey hair stimulation,but no thermal hyperalgesia was *** showed significant anti-allodynic effects on the inflammatory bone pain induced by our *** CFA is an effective agent for inducing bone inflammation and subsequent pain-related behavior in rat *** described rat model of inflammatory bone pain induced by CFA provides a practical and valuable contrast for bone cancer pain research.

关键词： inflammatory bone pain CFA mechanical allodynia celecoxib rat

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Multiple glial-neuronal dialogues in morphine tolerance:role of P2X7 receptor downstream cascades in the spinal cord

Multiple glial-neuronal dialogues in morphine tolerance:role...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Meng-Ling CHEN~#,Hong CAO~#,Long-Zhen CHENG,Yu-Xia CHU,Yu-Qiu ZHANG,Zhi-Qi ZHAO~* Institute of Neurobiology,Institutes of Brain Science & State Key Laboratory of Medical Neurobiology,Fudan University,Shanghai 200032,China

Objective Morphine is a potent ***,its clinical application is restricted due to morphine *** underlying mechanism is not fully *** are thought to be involved in morphine tolerance, and P2X purinergic receptor（P2XR） has been implicated in neuron- glia communication and chronic pain. This study investigated microglia-expressed P2X receptor（P2XR） and its downstream mechanisms underlying the induction of tolerance to morphine analgesia in *** Behavioral tests,SiRNA,Western Blotting, immunohistochemistry,ELISA and electrophysiology methods were used in this *** Chronic morphine treatment induced morphine tolerance and an increase in the expression of P2XR and IL-18 by microglia,IL-18 receptor（IL-18R） by astrocytes and PKCy by neurons in the spinal cord,respectively,which was blocked by a P2XR antagonist or targeting ***,the release of astroglial D-serine,a ligand for NMDA receptor,was increased by chronic morphine,which was attenuated by the D-serine *** D-serine-mediated presynaptic mechanism may contribute to the potentiation of spinal pain-sensitive neurons by chronic morphine treatment. Conclusion The present study revealed multiple dialogues between glial and neuronal cells via cascades involving P2XRs,IL-18,D-serine,NMDARs and PKCy in morphine tolerance,which may provide novel targets for preventing morphine tolerance.

关键词： morphine tolerance P2X7 receptor IL-18 D-serine PKCy glial-neuronal dialogue

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Neural stem cells in the SVZ niche interact with their neighbors

Neural stem cells in the SVZ niche interact with their neigh...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Qin SHEN Tsinghua University,Center for Stem Cell Biology and Regenerative Medicine

Neural stem cells(NSCs) are found in the adult brain only in specialized microenvironment or niches,mainly the subventricular zone(SVZ) of the lateral ventricle and the hippocampal dentate *** neural stem cell lineage in the SVZ includes type B cells,which express GFAP and contain the slow-dividing neural stem cells,Type C cells that are transit-amplifying cells,and type A neuroblasts which migrate anteriorly along the rostral migratory stream into the olfactory *** and lineage progression of NSCs are regulated by interplays between the NSC intrinsic properties and the stem cell niche *** previously showed that secreted factors from endothelial cells stimulate the self-renewal of neural stem cells and enhance their neurogenic potential in *** confocal imaging analysis,we characterized the 3-D cellular architecture of the SVZ niche,showing that adult SVZ NSCs and their progeny are bordered by the ependymal cells,which line the lateral ventricles,and blood vessels that form a plexus in the *** and progenitor cells are closely apposed to the laminin-containing extracellular matrix surrounding vascular endothelial *** work has identified a vascular niche in the SVZ and revealed an interaction between laminin and its receptor alpha6beta1 integrin which plays a functional role in binding SVZ stem cells within the vascular ***,apical GFAP-expressing cells are admixed in the ependymal layer and have contact with both the lateral ventricle and blood vessels,occupying a unique *** have further investigated molecules and signaling pathways regulating the placement and movement of NSCs in the SVZ niche,which will help understand how stem cell niche factors affect NSC activities and adult neurogenesis.

关键词： Neural stem cells in the SVZ niche interact with their neighbors niche

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Acute administration of fluoxetine normalizes rapid eye movement sleep abnormality,but not depressive behavior in olfactory bulbectomized rats

Acute administration of fluoxetine normalizes rapid eye move...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Zhi-Cai TU~1,Yi-Qun WANG~1,Rui LI~1,Xing-Yuan XU~1,Zhi-Li HUANG~(1,2,3,*) 1 Department of Pharmacology 2 State Key Laboratory of Medical Neurobiology,Shanghai Medical College 3 Institute of Brain Science,Fudan University,Shanghai 200032,China

Objective Depression in humans is associated with sleep abnormalities of three types:altered rapid eye movement(REM) sleep,fragmented sleep,and reduced delta ***,the cause and effects between depression and sleep abnormalities remain *** Bilateral olfactory bulbectomy(OBX) was carried out to make a depressive model in ***-wake profiles were assayed by the cutting-edge sleep bioassay system,and depressive behaviors were identified by open-field and forced swimming *** contents of monoamine and their metabolites in the brain were determined by ***(1) OBX rats exhibited a significant increase in REM sleep,especially the last 6-h during the light-on periods and attenuated non-REM sleep rebounds than sham rats after 6-h sleep deprivation(SD).(2) After acute treatment with fluoxetine(10 mg/kg,i.p.),the OBX-induced increase in REM sleep was abolished immediately,but the hyperactivity and the enhancement of immobility time remained unchanged.(3) Neurochemistry studies revealed that acute administration of fluoxetine increased the contents of serotonin(5-HT) in the hippocampus,thalamus and midbrain;decreased 5-hydroxyindoleacetic acid(5-HIAA) as well as the turnover of 5-HT(5-HIAA/5-HT) in almost all regions of the *** These results indicate that REM sleep abnormalities,but not the depressive behavior in OBX rats were reversed by acute administration of fluoxetine,and that there was no causality between REM abnormality and depression.

关键词： fluoxetine olfactory bulbectomy REM sleep serotonin

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Unconscious extraction of abstract auditory rules of speech:a mismatch negativity study using lexical tones

Unconscious extraction of abstract auditory rules of speech:...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Xiao-Dong WANG~(1,2),Feng GU~(1,2),Kang HE~(1,2),Ling-Hui CHEN~3,Lin CHEN~(1,2,*) 1 CAS Key Laboratory of Brain Function and Disease,School of Life Sciences,University of Science and Technology of China,Hefei 230027,China 2 Auditory Research Laboratory,University of Science and Technology of China,Hefei 230027,China 3 iFlytek Speech Laboratory,School of Information Science and Technology,University of Science and Technology of China,Hefei 230027,China

The abstract auditory rule of speech refers to the abstract and constant relations between speech stimuli, which has currently been under study for its underlying neural mechanisms at an attentive ***,it is unknown whether or not the human brain has a capacity of encoding abstract auditory rules of speech at a preattentive stage *** address this issue,we presented native Chinese speakers continuously with Chinese vowels differing in formant,intensity and level of pitch to construct an auditory input stream,in which most of vowels shared flat lexical tones to form an embedded abstract auditory rule,and occasionally this rule was violated with random occurrence of those with rising or falling lexical *** whole-head electrical recordings of the mismatch negativity,we show that the rule violation evoked a robust preattentive auditory response while none of the subjects acquired explicit knowledge of the rule or became aware of the violation of the *** results reveal a sensory intelligence in perception of Chinese lexical *** that Chinese lexical tones are semantically meaningful,our study further indicates the human's capability of unconscious extraction of abstract auditory rules of speech,which may be critical for speech communication in noisy and adversary auditory environments.

关键词： sensory intelligence lexical tone abstract auditory rule preattentive auditory response mismatch negativity

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Regulation of histone acetylation during consolidation of CPA extiction in vmPFC

Regulation of histone acetylation during consolidation of CP...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Wei-Sheng WANG~1,Shuo KANG~1,Wen-Tao LIU~2,Mu LI~1,Yao LIU~1,Chuan YU~1,Jing-Gen LIU~(1,*) 1 State Key Laboratory of Drug Research,Shanghai Institute of Materia Medica,Chinese Academy of Sciences, Shanghai 201203,China 2 School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210046,China

The drug-associated conditioned stimulus(CSs) come to influence drug-seeking and relapse behaviour through the memories they *** animals undergo extinction training(no drug) will decrease response rates and extinction entails new learning rather than *** the model of conditioned place aversion(CPA) of acute morphine-dependent rats,we found that lesions of ventral medial prefrontal cortex(vmPFC) impaired the consolidation of extinction of *** addition,we found that both extracellular signal-regulated kinases(ERK) pathway and histone acetylation in vmPFC are involved in consolidation of extinction and also found that Trichostatin A,a histone deacetylase(HDAC) inhibitor,can facilitate the extinction of *** further found that manipulation of vmPFC ERK activity by its antagonist U0126 and agonist D-cycloserine not only differentially influenced the behavior of CPA rats but also altered histone ***,we found that extinction of CPA increased BDNF exonⅠbut not exonⅣin the vmPFC,We further found that the increasement is accompanied by a significant increase in histone H3 acetylation and pCREB around the BDNF exon I gene *** also found that inhibition of trk B by intra-vmPFC injections of K252a significantly disrupted the consolidation of ***,our findings suggest that histone acetylation is mediated by ERK pathway and the increasement of BDNF protein expression in vmPFC are required for the consolidation of CPA extinction and the tyrosine kinase inhibitor K252a prevented the consolidation of CPA.

关键词： extinction vmPFC ERK histone acetylation BDNF

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Expression of Nurrl in rat brain and spinal cord development

Expression of Nurrl in rat brain and spinal cord development

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Weibo SHI,Bin CONG,Qian QI,Zuo MIN,Yingmin LI~* Institute of Basic Medicine,Hebei Medical University,Shijiazhuang 050017,China

Nurr1 is a member of the nuclear receptor superfamily and is a regulatory factor of differentiation, migration,and maturation of mesencephalic dopaminergic *** present study was designed to observe the dynamic changes of protein expression of Nurr1 as well as its relationship with proliferating cell nuclear antigen (PCNA) in the rat brain and spinal cord development,and to investigate the significance of Nurr1 in differentiation and migration of nerve ***-embedded sections,immunohistochemistry,immunohistochemical double staining and western blot were *** results demonstrated that in embryo development,the Nurr1-positive cells increased apparently and migrated gradually far from the lateral *** postnatal rats,the positive cells surrounding the lateral ventricle decreased *** expression of Nurr1 in the cerebral cortex peaked at days P1-P5,decreased obviously along with maturation of the cells and became rare in matured cerebral *** staircase-shaped migration from dorsal areas to ventral areas of spinal cord was ***,the positive cells in spinal cord decreased apparently,and there were no positive cells in matured spinal *** comparative observation of Nurr1 and PCNA showed that the two proteins were expressed at different regions and in different cells. Nurr1 was confined to differentiated and migrating immature cells,but not to proliferating *** suggested that Nurr1 might play a regulatory role in the differentiation,migration and maturation of the nerve cells in the rat brain and spinal cord.

关键词： Nurrl differentiation migration PCNA proliferation

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