成年大鼠液压冲击脑损伤诱导室下区Nestin和GFAP的表达

目的　应用免疫组织化学和免疫荧光双标技术结合激光共聚焦显微镜 ,观察脑损伤后不同时期室下区巢蛋白(Nestin)和胶质酸性纤维蛋白 (GFAP)的表达及其细胞类型。方法　用动液压冲击法建立动物模型 ,用LeicaQ5 0 0IW图像处理系统及免疫组化和免疫荧光双标法分析和显示不同时期 (1、3、5d ,1、2、3、4、6、8周 )Nestin和GFAP的表达变化。结果　损伤 2 4h后 ,室下区有大量Nestin和GFAP的表达 ,且激光共聚焦显微镜观察到有Nestin阳性突起与GFAP共存现象 ,7d左右达到高峰 ,然后随着时间的推移其表达逐渐下降。正常组和假手术组 ,室下区也可见到Nestin和GFAP的表达 ,但与脑损伤组相比较低 (P <0 .0 5 )。结论　脑损伤可诱导室下区Nestin和GFAP的表达 ,Nestin阳性突起与GFAP共存细胞可能具有干细胞特性 ,与神经元的修复有关。     

PROPERTIES OF PROLIFERATION AND DIFFERENTIATION OF NEONATAL RAT RETINAL PROGENITOR CELLS IN VITRO

Objective To investigate the properties of proliferation and differentiation of neonatal rat retinal progenitor cells （RPCs） in vitro. Methods RPCs were isolated from neonatal SD rats neural retina and cultured in DMEM/F12 ＋ N2 with EGF and bFGF （suspension medium ） or 10 % FBS without EGF and bFGF （ differentiation medium）. The cells grew as suspended spheres or adherent monolayers, depending on different culture conditions. The neural stem cells or retinal progenitors, neurons, astrocytes, retinal ganglion cells, rod photoreceptors and the proliferating cells were evaluated with immunofluorescence analysis by Nestin or Pax6, Map2, GFAP, Thy-1, Rhodopsin and BrdU antibodies respectively. Results RPCs could propagate and differentiate in suspension or differentiation medium and express the markers of Nestin （92.86%） or Pax6 （86.75%）, Map2 （38.54%）, GFAP （20.93%）, Thy-1 （27. 66%） and Rhodopsin（13. 33%）in suspension medium; however, Nestin （60.27%）, Pax6 （52%）, Map2 （34.94%）, GFAP （38.17%）, Thy-1 （30.84%） and Rhodopsin （34.67%） in differentiation medium. 96.4 % of the population in the neurospheres was BrdU-positive cells. The cells could spontaneously adherent forming some subspheres and retinal specific cell types. Conclusion Neonatal rat RPCs possess the high degree of proliferation and can differentiate into neurons, astrocytes, retinal ganglion cells and rod photoreceptors in vitro. There are different proportions for RPCs to differentiate into specific cell types.     

神经干细胞经脑室移植后在损伤脊髓的早期动态分布

目的观察神经干细胞经脑室移植后在损伤脊髓的早期动态分布。方法取转录有绿色荧光蛋白(GFP)基因的孕16 d SD鼠胚脑海马组织,培养成神经干细胞球,注射到脊髓损伤鼠第四脑室,通过免疫染色和电镜,观察其在脊髓的存活、分化状况。结果移植细胞在脊髓表面形成细胞团,大多分布于损伤区头侧的腹、背侧两面。细胞团面积背侧小于腹侧;数目背侧多于腹侧,并于术后5 d达到高峰。这种分布和增殖形式见于损伤脊髓正常部分和无损伤脊髓(对照组)。1周时细胞侵入损伤区,分化并表达神经胶质纤维酸性蛋白(GFAP)。2~3周时与宿主细胞良好融合。结论通过脑脊液移植细胞对于损伤性脊髓进行干细胞移植是一种实用、有效的方法。     

Identification and culture of neural stem cells isolated from adult rat subventricular zone following fluid percussion brain injury

Objective To analyze proliferation and differentiation of glial fibrillary acid protein (GFAP)- and nestin-positive (GFAP+/nestin+) cells isolated from the subventricular zone following fluid percussion brain injury to determine whether GFAP+/nestin+ cells exhibit characteristics of neural stem cells. Methods Male Sprague-Dawley rats, aged 12 weeks and weighing 200-250 g, were randomly and evenly assigned to normal control group and model group. In the model group, a rat model of fluid percussion brain injury was established. Five days later, subventricular zone tissue was resected from each group and made into single cell suspension. After serum-free neural stem cell medium culture and subsequent serum-induced differentiation, cell type, proliferation and differentiation capacities were determined by immunofluorescence staining and flow cytometry. Results At 3-7 days after fluid percussion brain injury, nestin+/GFAP+ cells in the single cell suspension from the model group significantly outnumbered those from the normal control group (P<0.01). In the model group, an increased number of small neurospheres with smooth cell edge and bulged center formed after primary culture, and were clearly visible with the increase of culture time and medium replacement. After several passages, many clonal spheres were obtained, suggesting strong self-proliferatiing capacity. Neurospheres from the model group differentiated into astrocytes, neurons and oligodendrocytes. Conclusion GFAP+/nestin+ cells isolated from the adult rat subventricular zone after fluid percussion brain injury are thought to be neural stem cells because of their self-renewal and multi-differentiation capacities.     

麻黄碱对脑缺血再灌注损伤大鼠脑内星形胶质细胞GFAP表达的影响

目的探讨大鼠脑缺血再灌注损伤后麻黄碱治疗对星形胶质细胞胶质原纤维酸性蛋白(GFAP)表达水平的影响。方法雄性SD大鼠60只,随机分为假手术组、自然恢复组和治疗组。应用线栓法建立单侧大脑中动脉缺血再灌注模型,术后1、2、3、4周应用免疫组化观察缺血周围区GFAP的表达。结果自然恢复组和治疗组术后1周GFAP表达开始增加,3周后表达平稳。治疗组GFAP的表达水平在前两个时间点高于自然恢复组(P<0.05)。结论麻黄碱对脑缺血再灌注损伤大鼠缺血周围区星形胶质细胞增殖活化有促进作用。     

大鼠脊髓损伤后MAP-2的表达及与运动功能恢复的相关性

目的研究大鼠脊髓损伤后MAP-2表达的变化及与运动功能恢复的相关性。方法健康成年Wistar大鼠36只,随机取6只作为正常对照组,余30只制作成脊髓打击伤动物模型,随机分为5组,每组6只。伤后1、3、7、14、28d取材,应用免疫组织化学方法观察MAP-2的表达,采用计算机图像分析系统,进行定量分析;用改良的Tarlov评分、斜板试验观察大鼠脊髓损伤后运动功能的恢复情况及MAP-2的表达和二者的相关程度。结果大鼠脊髓损伤后MAP-2的表达在损伤后1-14d呈进行性升高,并在损伤后14d达到高峰,损伤后28d较损伤后14d明显下降(P<0.01),但仍明显高于正常对照组(P<0.01),呈高水平表达;大鼠脊髓损伤后1-28d改良的Tarlov评分、斜板试验进行性升高,并在损伤后28d达到高峰,但仍明显低于正常对照组(P<0.01)。MAP-2的表达和改良的Tarlov评分、斜板试验在损伤后呈正相关(|r1|=0.81,P<0.01;|r2|=0.79,P<0.01)。结论大鼠脊髓损伤后MAP-2的表达和运动功能的恢复呈正相关,MAP-2很可能参与了大鼠脊髓损伤后运动功能的恢复。     

CORRELATION OF THE MAGNETIC MOTOR EVOKED POTENTIAL TECHNIQUE WITH PATHOLOGICAL CHANGES ASSOCIATED WITH SPINAL CORD INJURY IN ADULT CATS

Ｍａｇｎｅｔｉｃｍｏｔｏｒｅｖｏｋｅｄｐｏｔｅｎｔｉａｌｉｓａｎｏｎ－ｉｎｖａｓｉｖｅｅｌｅｃｔｒｏｐｈｙｓｉｏｌｏｇｉｃｔｅｃｈｎｉｑｕｅ,ｃｕｒｒｅｎｔｌｙｉｎｕｓｅ,ｉｎｔｈｅｃｌｉｎｉｃａｌｄｉａｇｎｏｓｉｓｏｆｔｈｅｅｘｔｅｎｔｏｆｓｐｉｎ...     

APOPTOSIS AFTER SPINAL CORD INJURY IN RATS

Traditionallythinking,themodeofneurondeathafterspinalcordinjuryisnecrosis.There-centstudiesshowthatapoptosismayplayaroleinthepathophysiologicalprocessofspinalcordin-jury"'-Inthisstudy,apoptosisinspinalcordin-juryinratswasobsevedandanalyzedinordertooffertheoreticalevidenceforpathogenesisofspinalcordinjury.MATERIALSANDMETHODS1Animalsurgeryandtissueprocessing36femaleSprague-Dawleyratsweighing25ol2Ogwererandomlydividedinto9groups,4ineachgroup.onegroupconsistedofnormalnon-operatedrats,inth…     

The β-amyloid protein induces S100β expression in rat hippocampus through a mechanism that involves IL-1

Objective To explore the effect of β-amyloid protein (Aβ) on S100β expression in rat hippocampus and its mechanisms. Methods At 7 days after bilateral stereotaxis injection of different dose of fibrillar Aβ 25-35 and interluekin-1 receptor antagonist (IL-1ra) into the rat CA1 region, the learning and memory abilities of rats were tested with passive avoidance task. Amyloid deposition was detected by using Congo red staining technique. Nissl staining and immunohistochemical techniques were used to analyze the number of neurons, and GFAP and the S100β expression in hippocampal CA1 region , respectively. Results After fibrillar Aβ injection, the step-through latency of rats was significantly shortened compared to that of the control group. The GFAP positive astrocytes were found surrounding amyloid deposition. Neuronal loss occurred in the pyramidal cell layer of CA1 region. The number of S100β positive cells in Aβ-treated group was significantly increased compared with that in the control group. After IL-1ra injection, the number of S100β positive cells was significantly decreased. Conclusion Intrahippocampal injection of Aβ 25-35 could cause similar pathologic changes of Alzheimer's disease. Aβ 25-35 was capable of up-regulating S100β expression in a dose-dependent manner. The injection of IL-1ra could attenuate the effect of Aβ on S100β expression.     

EXPERIMENTAL STUDY ON SPINAL CORD INJURY TREATED WITH THE COMBINATION OF FETAL SPINAL CORD TRANSPLANTATION AND METHYLPREDNISOLONE

ＴｈｅｔｈｅｒａｐｙｏｆＬａｒｇｅＤｏｓａｇｅｏｆＭｅｔｈｙｌｐｒｅｄｉｎ ｓｏｌｏｎｅ(ＭＰ)ｆｏｒＳＣＩｈａｓｄｅｖｅｌｏｐｅｄｃｌｉｎｉｃａｌｌｙｂｅｃａｕｓｅｉｔｃａｎｇｒｅａｔｌｙｉｍｐｏｖｅｔｈｅｎｅｒｖｅｆｕｎｃｔｉｏｎ .Ｉｎｔｈｅｍｅａｎ ｔｉｍｅ,ａｎｉｍａｌｔｅｓｔｓｈａｖｅａｌｒｅａｄｙｆｏｕｎｄｔｈａｔｉｆｆｅｔａｌｓｐｉｎａｌｃｏｒｄｉｓｔｒａｎｓｐｌａｎｔｅｄｔｏｔｈｅｉｎｊｕｒｅｄｓｐｉｎａｌｃｏｒｄａｒｅａ ,ｉｔｃａｎｓｔｉｍｕｌａｔｅｔｈｅａｘｏｎｓｏｆｔｈｅｈｏｓ…     

大鼠急性脊髓损伤中caspase-1的表达及意义

目的研究半胱氨酸天冬氨酸特异性蛋白酶-1(cysteinyl aspartate specific protease-1,caspase-1)在大鼠急性脊髓损伤后的表达及意义。方法取健康成熟SD大鼠36只,随机分为对照组和损伤组,各18只,每组再分8 h、3 d7、d共3个时间点,每个时间点6只。对照组仅做单纯T8、T9椎板切除术,损伤组则按Nystrom法建立大鼠急性脊髓损伤动物模型。HE染色观察脊髓组织病理学变化,免疫组化测定各时间点caspase-1的表达变化。结果对照组大鼠脊髓组织中少量细胞caspase-1表达阳性。脊髓损伤后8 h时,损伤组caspase-1表达高于对照组,3 d时表达最高,7 d时略有降低,但仍高于对照组。脊髓损伤组各时间点caspase-1表达阳性细胞数较对照组明显升高(P<0.01)。结论大鼠脊髓损伤后caspase-1的表达迅速增强,可能是导致脊髓继发性损伤的因素之一。     

EFFECTS OF CEREBRAL CORTICAL CONCIS ON CELL PROLIFERATION OF THE SUBVENTRICULAR ZONE IN ADULT RATS

Mechanical injuries to the external regions ofthe brain including the cerebral cortex and otherparts of the telencephalon are common yet relativelyuntreatable[1].The predicament in recovery frombrain injury is that the adult central nervous systemis generally thought to be incapable of replacingdead neurons.As the SVZis nowknownto be neu-rogenic andis in close proxi mitytothe cerebral cor-tex and other functionally i mportant forebrain nu-clei,hope has risen that its neurogenesis may be co-op…     

PROTECTIVE EFFECTS OF LIGUSTRAZINE ON ISCHEMIA-REPERFUSION INJURY IN RAT KIDNEYS

Ｄｕｒｉｎｇｒｅｎａｌｉｓｃｈｅｍｉａ ｒｅｐｅｒｆｕｓｉｏｎｉｎｊｒｕｙ ,ｎｕ ｍｅｒｏｕｓｆｒｅｅｒａｄｉｃａｌｓａｒｅｐｒｏｄｕｃｅｄａｎｄＭＤＡ ,ａｐｒｏｄ ｕｃｔｏｆｆｒｅｅｒａｄｉｃａｌ,ａｎｄＥＴ 1ｉｎｋｉｄｎｅｙｓｂｏｔｈｉｎｃｒｅａｓｅｉｎｔｈｅｐｒｏｃｅｓｓｏｆｒｅｐｅｒｆｕｓｉｏｎ .ＳＯＤｉｓｃａｂｌｅｏｆｅｌｉｍｉ ｎａｔｉｎｇｆｒｅｅｒａｄｉｃａｌｓ .ＡｄｍｉｎｉｓｔｒａｔｉｏｎｏｆＳＯＤ ｐｒｉｏｒｔｏｉｓｃｈｅｍｉａｏｒａｔｔｈｅｂｅｇｉｎｎｉｎｇｏｆｒｅｐｅｒｆｕｓｉｏ…     

CALCIUM ANTAGONIST AND ENDOTHELIN IN THE ACUTE SPINAL INJURY

Acutespinalinjury(ASI)isakindofdiseasecommonlyseeninthedepartmentoforthopedics,withsecondarypathologicinjurycausingthede-layeddamageoftissuestructure.Manybiopatho-giccausespla_y_roles1nthecomplexcourse,how-ever,intrace11ularcalciumoverloadwasregardedasthemajordamageofthesecondaryinjuryofSCl.Withthedevelopmentofendotheliumbioac-tivitystudy,itseffectsonthecentralnervoussys-temdrawmoreandmoreattention.Andtherewerenosystematicreportsontherelationbetweencalciumandendotheliumt1.2i.Thegoalofthisstu…     

重症急性胰腺炎胰腺损伤与肺损伤的关系

目的探讨重症急性胰腺炎(SAP)引起急性肺损伤(ALI)的机制。方法48只实验组大鼠注射40g/L(4%)牛磺胆酸钠制作SAP模型。实验组及对照组(48只)大鼠分别在1、3、5、7、9、12h点剖杀,取胰腺及肺组织行病理观察、微血管通透性及MPO活性测定;RT-PCR法检测胰腺及肺组织中细胞间黏附分子(ICAM-1)、肿瘤坏死因子(TNF-α)基因表达。结果制模后胰腺及肺损伤逐渐加重,胰腺及肺髓过氧化物酶(MPO)活性及微血管通透性逐渐升高;胰腺TNF-α、ICAM-1基因表达均在1h明显升高,至7h达到峰值;肺组织TNF-α、ICAM-1基因表达则在1h后逐渐上升,至9~12h达到峰值。结论SAP引起ALI过程中存在明显的时间窗口,该窗口内的早期预防性保护可能利于防止ALI的发生。     

神经递质P物质通过调控转录因子Osterix的表达促进成骨细胞分化

目的 研究神经递质P物质调控成骨细胞分化的分子途径.方法 分离骨髓基质干细胞进行原代及传代培养;分别采用空白对照、P物质、P物质NK1受体拮抗剂、P物质+P物质NK1受体拮抗剂进行干预,诱导骨髓基质干细胞向成骨细胞分化;传代培养1～2周后,抽提细胞总RNA,用RT-PCR检测分化过程中Osterix基因的表达.检测结果重复3次,采用单因素方差分析检测结果.结果 骨髓基质干细胞在生长对数增殖期为4～6 d,采用RT-PCR检测发现P物质干预成骨细胞分化,导致成骨细胞分化过程中重要的转录引子Osterix 基因表达,与其他各组比较有显著差异(P<0.05),Osterix基因表达上调,从而刺激前成骨细胞向成骨细胞转化.而P物质+P物质NK1受体拮抗剂共同干预,Osterix基因表达与空白对照组无显著差异(P<0.05),说明P物质通过P物质NK1受体对成骨细胞分化进行调控.结论 P物质可调控前成骨细胞分化过程中转录因子Osterix基因表达促进其向成骨细胞分化.P物质对Osterix基因表达的调控依赖P物质NK1受体.     

β-淀粉样蛋白诱导大鼠海马S100β表达的作用及机制

目的　探讨 β 淀粉样蛋白 (Aβ)对S10 0 β表达的影响及作用机制。 方法　采用Aβ2 5-3 5和IL 1ra ,选择大鼠海马CA1区进行定位注射 ,通过行为学测试、刚果红染色和免疫组化技术结合图像分析的方法 ,对不同时间大鼠的学习记忆、淀粉样沉积、GFAP和海马CA1区S10 0 β表达的变化进行观测。 结果　Aβ2 5-3 5注射后 ,大鼠的学习记忆能力明显下降 ;海马CA1区出现Aβ沉积 ,并有大量GFAP阳性胶质细胞包绕 ;实验组S10 0 β阳性细胞数目在 3、7、2 1d较对照组均有明显增多 ,细胞平均面积只在 2 1d才有明显增大。脑内注射IL 1ra后 3、7d,S10 0 β阳性细胞数目明显低于同组的Aβ大鼠 ,而高于对照组。结论　Aβ2 5-3 5脑内注射可建立具有类似阿尔茨海默病的局部病理改变和学习记忆损害的AD动物模型 ;Aβ2 5-3 5可上调大鼠海马CA1区S10 0 β的表达 ,实验早期以S10 0 β阳性细胞的数目增加为主 ,后期表现为细胞体积的增大 ;IL 1ra脑内注射可明显降低Aβ2 5-3 5上调S10 0 β表达的作用 ,其机制是阻断由Aβ2 5-3 5激活的小胶质细胞释放的IL 1对星形胶质细胞的活化作用。     

ROLE OF ANTISENSE AND DECOY OLIGONUCLEOTIDE OF NF-κB IN VESSEL STENOSIS AND NEOINTIMA FORMATION IN BALLOON-INJURED RAT ARTERY

Objective To examine antisense and decoy oligonucleotides of nuclear factor kappa B in vivo effects on intima proliferation and balloon-injured monocytes chemotactic protein-1 ( MCP-1 ) and extracellular signal regulated kinase-2 ( ERK2 ) expression in the carotid artery of rats. Methods Sprague-Dawley rats underwent balloon-dilation injury of the left carotid artery. Rats are divided into 7 groups ( n = 18 ) and each group includes 6 time points (6 h, 1, 3, 5, 7, 14 d) ( n =3). Uninjured right carotid artery of the same rat was used as controls. Results In model group, sense group and scramble group, vessel intima area , media area and intima/media ratio increased after 5 d and reached the maximum after 7 d. The effect of antisense plus decoy group on intimal hyperplasia was more obvious than that of antisense group and decoy group alone. MCP-1 mRNA expression was increased expression continuously at 3, 5 and 7 d and decreased at 14 d. Compared with model group, sense group and scramble group, antisense group, decoy group and antisense plus decoy group had lowered MCP-1 mRNA expression in each time point ( P ＜ 0. 05 ). NF-κB p65 was dispersed positive stain 6 h after injury and increased after 1 d and peaked at 7 d, but the protein expression was weak at 14 d. ERK2 protein synthesis increased at 1 d and reached the peak at 7 d, while protein expression after 14 d was similar to that at 7 d. Treatment of antisense group,decoy group and antisense plus decoy group inhibited protein synthesis more significantly than those of model group,sense group and scramble group ( P ＜ 0. 05 ). Conclusion NF-κB modulates genes expression and protein synthesis of MCP-1 and ERK2. Celluar proliferation in vessel wall was dynamically changed after balloon angioplasty injury. Antisense and decoy oligonucleotide of NF-κB by local lipofectamine transfer inhibit NF-κB activating gene modulation and neointimal hyperplasia.     

脊髓损伤后大鼠后肢运动功能恢复不同评分标准的比较

目的比较脊髓损伤(SCI)后大鼠后肢运动功能恢复的不同评分标准的优劣。方法40只SD成熟雌性大鼠随机分为正常组(Normal组)、急性脊髓中度损伤组(SCI组)及对照组(CON组),其中SCI组采用改良的Allen打击法,CON组仅行T10椎板切除术。术后1、2、3、4、6周观察大鼠后肢神经功能恢复的情况并记录结果。评价标准分别为:斜板试验评分、改良Tarlov评分及BBB评分。结果SCI组与CON组比较,斜板试验临界角度在1-6周时,均有所减小(P<0.05),尤以第1周时减少更甚(P<0.01);改良Tarlov评分第1、2、3、4周时,分值间的差别非常明显(P<0.01),第6周时,未见变化(P>0.05);而BBB评分各时间点的区分程度非常明显(P<0.01)。结论BBB评分对SCI模型运动功能评价具有明显优势,可作为今后研究的标准评分法。     

EFFECTS OF NERVE GROWTH FACTOR ON ENDOTHELIN AFTER SPINAL CORD INJURY IN RATS

Ｗｉｔｈｔｈｅｄｅｖｅｌｏｐｍｅｎｔｏｆｍｅｄｉｃｉｎｅ ,ｓｐｉｎａｌｃｏｒｄｉｎｊｕｒｙ (ＳＣＩ)ｈａｖｅｂｅｅｎｔｈｅｍｏｓｔｄａｎｇｅｒｏｕｓｆａｃｔｏｒｆｏｒｔｈｅｈｅａｌｔｈ .ＢｕｔｎｅｒｖｅｇｒｏｗｔｈｆａｃｔｏｒｍａｙｂｅｔｈｅｈｏｐｅｏｆｔｒｅａｔｍｅｎｔｆｏｒＳＣＩ .Ｗｅｏｂｓｅｒｖｅｄｔｈｅｃｈａｎｇｅｓｏｆｅｎ ｄｏｔｈｅｌｉｎ(ＥＴ)ｓｅｒｕｍｌｅｖｅｌａｆｔｅｒｔｈｅｉｎｊｕｒｙｏｆｓｐｉｎａｌｃｏｒｄ ,ｔｈｅｒｅｓｕｌｔｓｗｅｒｅｒｅｐｏｒｔｅｄｈｅｒｅｔｏｉｎｖｅｓｔｉ…