胀亡-持续性局灶脑缺血星形胶质细胞的死亡方式

目的动态观察大鼠持续性局灶脑缺血后星形胶质细胞的形态学变化,探讨星形胶质细胞的死亡方式.方法将48只SD大鼠随机分为免疫组化组、免疫组化对照组、电镜实验组和电镜实验对照组.采用颈内动脉线栓并环扎的方法建立持续性局灶脑缺血模型.免疫组化组和电镜实验组大鼠按照缺血时间（3h、6h、12h、24h、48h）平均随机分为5个亚组.对照组为术后24h取脑.免疫组化标本采用HE染色及TUNEL-GFAP双染.电镜标本在中心区、边缘区分别取材、染色后,透射电镜观察.结果随着时间的延长,脑缺血中心区及边缘区星形胶质细胞逐渐出现核肿胀、染色质分散、边聚、空泡变,核膜破溃,染色质溢出.随着缺血时间延长,中心区GFAP阳性细胞数逐渐减少,直至基本消失.缺血边缘区则可见到GFAP阳性细胞数量逐渐增多.TUNEL-GFAP双染未见阳性细胞.结论持续局灶脑缺血后,星形胶质细胞未见凋亡,推测胀亡是星形胶质细胞主要的死亡方式.     

大鼠持续脑缺血后星形胶质细胞的时空分布及其变化

目的 本实验研究星形胶质细胞在持续脑缺血后的时空分布及其变化。方法 选用SD雄性大鼠54只，随机分为分成假手术组和手术组。手术组又按缺血时间（3、6、12、24及48 h）分为5个亚组。假手术组和每亚组各9只大鼠。采用颈内动脉线栓加环扎法复制大鼠持续脑缺血模型，相应时间点灌注取脑，常规固定包埋切片，HE和TdT介导的dUTP缺口末端标记（TUNEL）、胶质纤维酸性蛋白（GFAP）染色。结果 随着持续脑缺血时间的延长，中心区神经细胞及星形胶质细胞的数量逐渐减少，缺血边缘区凋亡的神经细胞数量逐渐增多，而星形胶质细胞在12 h达到最低点之后，数量逐渐增多，并呈空泡状改变。结论 在持续缺血12 h之后，缺血中心区与半暗带区星形胶质细胞数量变化不一致，在半暗带其数量逐渐增多，提示星形胶质细胞在缺血后的不同阶段，其作用可能不同。     

脑室注射痴呆大鼠海马胶质纤维酸性蛋白表达与学习记忆能力的关系

目的探讨星形胶质细胞在老年性痴呆大鼠海马中的表达与老年性痴呆大鼠学习记忆能力减退的关系。方法雄性SD大鼠20只,随机分为痴呆组与假手术组;用Morris水迷宫检测大鼠的学习、记忆能力;用免疫组化技术定量检测大鼠海马CA1区胶质纤维酸性蛋白(GFAP)的表达;分析海马星形胶质细胞变化与学习、记忆能力的关系。结果假手术组海马CA1区锥体细胞排列紧密有序,细胞核大而圆、染色浅、核仁明显、未见明显胞浆浓染、核固缩等神经元变性受损征象。而痴呆大鼠海马CA1锥体神经细胞排列疏松、数目减少、细胞形态异常,许多细胞出现体积缩小、核浓染、核固缩;痴呆鼠海马CA1区GFAP阳性细胞数目明显增多,胞体肥大,突起增粗、变长现象明显,而假手术组海马CA1区仅见少量GFAP阳性细胞,突起较少、短,染色较淡。计数和测量海马CA1区GFAP阳性细胞数目、总面积、平均光密度,痴呆组与假手术组相比均明显增加,有显著意义(P<0.05);水迷宫测试显示痴呆组大鼠隐藏平台获得时间比假手术组明显延长,空间探索时间明显缩短,具有显著意义(P<0.05);显示痴呆组大鼠学习记忆能力与假手术组比较均明显下降;将痴呆大鼠学习成绩与海马CA1区GFAP表达数目之间进行相关分析,两者间存在负相关关系,认为星形胶质细胞参与了学习记忆过程。结论提示海马星形胶质细胞的过度表达可能影响痴呆大鼠的学习记忆能力。     

雪莲提取物对脑缺血再灌注损伤小鼠海马区星形胶质细胞胶质纤维酸性蛋白和小胶质细胞离子钙接头蛋白的影响

目的探讨雪莲提取物对缺血再灌注损伤小鼠海马区星形胶质细胞胶质纤维酸性蛋白(GFAP)和小胶质细胞离子钙接头蛋白(Iba1)表达的影响。方法 40只昆明小鼠随机分为假手术组、缺血再灌注组和雪莲提取物低、中、高剂量组。采用线栓法制作大脑中动脉闭塞模型。雪莲提取物低、中、高剂量组小鼠术前7 d分别连续腹腔注射0.2 g/(kg·d)、0.4 g/(kg·d)、0.8 g/(kg·d)雪莲注射液。假手术组不闭塞大脑中动脉,给予等体积中剂量雪莲注射液。缺血再灌注组再灌时腹腔注射等体积生理盐水。再灌注24 h后采用免疫组化染色法检测GFAP和Iba1的表达。结果与假手术组比较,缺血再灌注组及雪莲提取物低、中剂量组GFAP阳性细胞数明显增加,缺血再灌注组及雪莲提取物低、中、高剂量组IBA1阳性细胞数明显增加(均P0.01);与缺血再灌注组比较,雪莲提取物低、中、高剂量组GFAP及IBA1阳性细胞数明显减少(均P0.01);与雪莲提取物低剂量组比较,雪莲提取物高剂量组GFAP阳性细胞数明显减少,雪莲提取物中、高剂量组IBA1阳性细胞数明显减少(P0.05~0.01)。结论雪莲提取物可抑制缺血再灌注损伤小鼠海马区星形胶质细胞GFAP和小胶质细胞Iba1的过度表达,有神经保护作用。     

大鼠轻型颅脑损伤后星形胶质细胞与神经元的病理改变

目的 探讨轻型颅脑损伤（TBI）后神经元及星形胶质细胞改变的病理生理过程。方法 将24只成年SD大鼠随机分为轻型TBI组（n=18）和假手术组（n=6）,轻型TBI组又分为伤后3 h（n=6）、伤后24 h（n=6）、伤后72 h（n=6）三亚组。采用液压冲击法制作轻型TBI模型。采用胶质纤维酸性蛋白（GFAP）染色检测星形胶质细胞,采用Fluoro-Jade B（FJ-B）荧光染色检测变性神经元。结果 与假手术组相比,轻型TBI后3 h、24 h、72 h邻近顶叶皮质、海马CA2/3区GFAP阳性细胞数量均明显减少（P<0.05）;缺失区周围星形胶质细胞肿胀增生明显。FJ-B阳性神经元在损伤后3 h无明显增加（P>0.05）,伤后24 h皮层区FJ-B阳性神经元显著增加（P<0.05）,伤后72 h海马区FJ-B阳性神经元显著增加（P<0.05）。伤后72 h伤侧皮层区与海马区GFAP阳性细胞数和FJ-B阳性细胞数呈显著负相关（r=-0.8285,P<0.05）。结论 轻型TBI后星形胶质细胞超急性期（3 h）即出现损害和胶质反应,神经元则在急性期（24 h）至亚急性期（72 h）出现明显损害,星形胶质细胞缺失程度可以反应神经元损伤程度。     

大鼠全脑缺血再灌注后额叶神经细胞凋亡与P53蛋白表达的实验研究

目的 探讨大鼠全脑缺血再灌注后不同时间对额叶神经细胞凋亡及P~(53)蛋白表达的影响。方法采用改良的Pulsineli 4-血管阻断(4-VO)方法建立SD大鼠急性全脑缺血模型,随机分为3组:正常组(n=7);假手术组(n=49);手术组(n=49)。缺血15min,分别于再灌注1、6、12、24、48、72h和7d断头取脑,采用TUNEL方法检测神经细胞凋亡,SP免疫组化方法观察额叶P~(53)蛋白的表达。结果 全脑缺血再灌注24h,可见少量TUNEL阳性细胞,再灌注48h可见较多TUNEL阳性细胞,72h出现大量TUNEL阳性细胞,7d明显减少。免疫组化染色:缺血组于再灌注24h可见少量P~(53)蛋白表达,48h达高峰,72h有所下降,7d明显下降,这种表达主要在细胞核内。结论 急性全脑缺血再灌注后的迟发性神经元坏死是以凋亡的方式发生的,全脑缺血再灌注后,额叶P~(53)蛋白表达增加,神经细胞凋亡和P~(53)蛋白的表达在一定时间内呈正相关。     

大鼠全脑缺血再灌注后额叶神经细胞凋亡与P^53蛋白表达的实验

目的 探讨大鼠全脑缺血再灌注后不同时间对额叶神经细胞凋亡及P^53蛋白表达的影响。方法 采用改良的Pulsineli 4-血管阻断(4-VO)方法建立SD大鼠急性全脑缺血模型，随机分为3组：正常组（n=7)；假手术组（n=49)；手术组（n=49)。缺血15min，分别于再灌注1、6、12、24、48、72h和7d断头取脑，采用TUNEL方法检测神经细胞凋亡，SP免疫组化方法观察额叶P^53蛋白的表达。结果 全脑缺血再灌注24h，可见少量TUNEL阳性细胞，再灌注48h可见较多TUNEL阳性细胞，72h出现大量TUNEL阳性细胞，7d明显减少。免疫组化染色：缺血组于再灌注24h可见少量P^53蛋白表达，48h达高峰，72h有所下降，7d明显下降，这种表达主要在细胞核内。结论 急性全脑缺血再灌注后的迟发性神经元坏死是以凋亡的方式发生的，全脑缺血再灌注后，额叶P^53蛋白表达增加，神经细胞凋亡和P^53蛋白的表达在一定时间内呈正相关。     

二咖啡酰奎宁酸对脑缺血后星形胶质细胞Porimin指标的影响观察

目的观察二咖啡酰奎宁酸对脑缺血后星形胶质细胞Porimin指标的影响。方法 32只雄性SD健康清洁级大鼠,分为空白组、对照组及药物组。应用大脑中动脉阻塞模型(MCAO)在对照组和药物组诱导脑缺血发作后,在对照组中予生理盐水,在药物组中予二咖啡酰奎宁酸。在MCAO后6h、24h分别进行行为学神经功能评分,HE染色、Cresyl Violet染色和GFAP、Porimin、DAPI免疫荧光染色。结果对照组和药物组中HE染色及Cresyl Violet均显示缺血中心区脑组织坏死,缺血半暗带区细胞水肿,可见肿胀的星形胶质细胞,对照组细胞损伤重于药物组。免疫荧光检测示药物组Porimin与GFAP、DAPI三标阳性细胞数目显著低于对照组(P=0.01)。结论二咖啡酰奎宁酸能降低脑缺血后星形细胞porimin指标的表达,提示二咖啡酰奎宁酸在脑缺血时对星形胶质细胞起一定积极的保护作用。     

大鼠脑缺血后海马CA1区胶质纤维酸性蛋白表达与迟发性神经元死亡的关系

目的观察大鼠大脑缺血再灌注后海马CA1区胶质纤维酸性蛋白(GFAP)的表达与迟发性神经元死亡的关系。方法采用大鼠大脑中动脉阻塞再灌注模型(MCAO),将大鼠随机分为MCAO后3d、7d、30d组及假手术组,应用免疫荧光与TUNEL染色法分别观察脑缺血再灌注后不同时间点缺血侧海马CA1区GFAP表达情况和迟发性神经元死亡(DND)的变化。结果(1)3d组海马DND阳性(DND 组)的MCAO大鼠、海马DND阴性(DND-组)的MCAO大鼠与假手术组大鼠比较,缺血侧海马CA1区GFAP染色的平均光密度无显著性差异(P>0.05),但GFAP阳性细胞的形态发生变化;(2)7d组大鼠缺血侧海马CA1区GFAP阳性细胞大量活化增殖,表现为胞体变大,突起增多;DND( )、DND(-)组海马CA1区GFAP染色的平均光密度较假手术组增高(P<0.01),且DND(-)组的GFAP平均光密度较DND( )组明显增高(P<0.01);(3)30d组大鼠缺血侧海马CA1区GFAP表达呈瘢痕样改变,DND( )、DND(-)组与假手术组比较其GFAP染色的平均光密度明显增高(P<0.05),且DND( )组的GFAP平均光密度较DND(-)组明显增高(P<0.05)。结论大鼠MCAO后星形胶质细胞反应性变化的差异可能与海马CA1区迟发性神经元死亡的发生有关。     

缺血缺氧对体外培养星形胶质细胞细胞周期和增殖的影响

目的 观察缺血缺氧损伤对星形胶质细胞细胞周期和增殖的影响。方法 用流式细胞仪检测缺血缺氧后不同时问点星形胶质细胞细胞周期变化，并用荧光免疫细胞化学技术测定胶质细胞纤维酸性蛋白(GFAP)和增殖细胞核抗原(PCNA)的表达水平。结果 体外缺血缺氧损伤后星形胶质细胞S期较正常组明显增高，6h达高峰，而随后则呈下降趋势。PCNA阳性反应损伤后表达均增加，6h表达最高；在缺血缺氧早期，GFAP阳性染色增强，6h最高；缺血缺氧12h后GFAP阳性染色变弱。结论 缺血缺氧损伤后星形胶质细胞活化进入增殖期；PCNA参与了损伤后星形胶质细胞的修复和增殖；细胞周期事件与星形胶质细胞的活化密切相关。     

大鼠脑干前庭核团向大脑前庭皮层的投射研究

目的观察Wistar大鼠脑干前庭核团是否存在直接向大脑皮质的纤维投射。方法健康Wistar大鼠20只,随机分为实验组(10只)和对照组(10只)。实验组脑干前庭内、外侧核团注射绿色荧光标记的顺行示踪剂刀豆凝集素,对照组脑干前庭内、外侧核团注射生理盐水。5d后处死大鼠,行大脑连续冰冻切片,荧光显微镜下观察荧光细胞在大脑皮质的分布及形态。结果绿色荧光标记的神经元主要位于大脑皮质的前肢感觉区、后肢感觉区和本体感觉区。结论 Wistar大鼠脑干前庭核团存在直接向大脑皮质的纤维投射,部分和本体感觉区相重叠。     

Effect of lidocaine on retinal aquaporin-4 expression after ischemia/reperfusion injury in the rat★

BACKGROUND: Several studies have demonstrated that high doses of lidocaine can reduce edema in rats with brain injury by down-regulating aquaporin-4 (AQP4) expression. The hypothesis for the present study is that lidocaine could retinal edema that is associated with AQP4 expression.OBJECTIVE: This study was designed to investigate the interventional effects of lidocaine on retinal AQP4 expression and retinal edema following ischemia/reperfusion injury in the rat.DESIGN, TIME AND SETTING: This study, a randomized, controlled, animal experiment, was performed at the Basic Research Institute, Chongqing Medical University from September 2006 to May 2007.MATERIALS: Seventy-five, healthy, adult, female, Sprague-Dawley rats were included. A total of 50 rats were used to establish a retinal ischemia/reperfusion injury model using an anterior chamber enhancing perfusion unit. Rabbit anti-rat AQP4 antibody was purchased from Santa Cruz Biotechnology, USA.METHODS: All 75 rats were randomly divided into three groups, with 25 rats in each: control, model, and lidocaine. At each time point (1, 6, 12, 24, and 48 hours after modeling, five rats for each time point), each rat in the lidocaine group was intraperitoneally administered lidocaine with an initial dose of 30 mg/kg, followed by subsequent doses of 15 mg/kg every six hours. The entire treatment process lasted three days for each rat. At each above-mentioned time point, rats in the model group were modeled, but not administered any substances. Rats in the control group received the same treatments as in the lidocaine group except that lidocaine was replaceld by physiological saline.MAIN OUTCOME MEASURES: Following hematoxylin-eosin staining, rat retinal tissue was observed to investigate retinal edema degree through the use of an optical microscope and transmission electron microscope. Retinal AQP4 expression was determined by immunohistochemistry.RESULTS: At each above-mentioned time point, AQP4 expression was significantly increased in the model group compared to the control group (P<0.05); this change was consistent with the degree of retinal edema. In the lidocaine group, retinal AQP4 expression was significantly decreased (P<0.05), and retinal edema was reduced, compared with the model group.CONCLUSION: Lidocaine inhibits rat retinal AQP4 expression following ischemia/reperfusion injury, leading to a reduction of retinal edema.     

Effect of lidocaine on retinal aquaporin-4 expression after ischemia/reperfusion injury in the rat★

BACKGROUND: Several studies have demonstrated that high doses of lidocaine can reduce edema in rats with brain injury by down-regulating aquaporin-4 (AQP4) expression. The hypothesis for the present study is that lidocaine could retinal edema that is associated with AQP4 expression. OBJECTIVE: This study was designed to investigate the interventional effects of lidocaine on retinal AQP4 expression and retinal edema following ischemia/reperfusion injury in the rat. DESIGN, TIME AND SETTING: This study, a randomized, controlled, animal experiment, was performed at the Basic Research Institute, Chongqing Medical University from September 2006 to May 2007. MATERIALS: Seventy-five, healthy, adult, female, Sprague-Dawley rats were included. A total of 50 rats were used to establish a retinal ischemia/reperfusion injury model using an anterior chamber enhancing perfusion unit. Rabbit anti-rat AQP4 antibody was purchased from Santa Cruz Biotechnology, USA. METHODS: All 75 rats were randomly divided into three groups, with 25 rats in each: control, model, and lidocaine. At each time point (1, 6, 12, 24, and 48 hours after modeling, five rats for each time point), each rat in the lidocaine group was intraperitoneally administered lidocaine with an initial dose of 30 mg/kg, followed by subsequent doses of 15 mg/kg every six hours. The entire treatment process lasted three days for each rat. At each above-mentioned time point, rats in the model group were modeled, but not administered any substances. Rats in the control group received the same treatments as in the lidocaine group except that lidocaine was replaceld by physiological saline. MAIN OUTCOME MEASURES: Following hematoxylin-eosin staining, rat retinal tissue was observed to investigate retinal edema degree through the use of an optical microscope and transmission electron microscope. Retinal AQP4 expression was determined by immunohistochemistry. RESULTS: At each above-mentioned time point, AQP4 expression was significantly increased in the model group compared to the control group (P < 0.05); this change was consistent with the degree of retinal edema. In the lidocaine group, retinal AQP4 expression was significantly decreased (P < 0.05), and retinal edema was reduced, compared with the model group. CONCLUSION: Lidocaine inhibits rat retinal AQP4 expression following ischemia/reperfusion injury, leading to a reduction of retinal edema. Key Words: ischemia/reperfusion; lidocaine; aquaporin-4     

狗枣猕猴桃叶提取物黄酮对大鼠局灶性脑缺血损伤组织病理结构的影响

目的研究提取物黄酮对局灶性脑缺血大鼠损伤组织病理及其超微结构的影响。方法Nagasawa线栓法将大鼠制成局灶性大脑中动脉缺血模型。将Wistar雄性大鼠75只随机分为:假手术组、盐水对照组、小剂量给药组、中剂量给药组、大剂量给药组,每组15只。小剂量给药组、中剂量给药组、大剂量给药组于术前30min及术后2h分别给予不同剂量狗枣猕猴桃叶提取物黄酮腹腔内注射。应用TTC染色及图像分析系统测定局部脑缺血24h后的梗死灶体积,HE染色及透射电镜观察大鼠局灶脑缺血后组织病理及超微结构的变化。结果与盐水对照组相比,药物干预组局部缺血后在相同时间内梗死灶的体积较小(P<0.05),超微结构改变也较轻,在实验剂量范围内其病理变化与给药剂量呈负相关。结论狗枣猕猴桃叶提取物黄酮对缺血后大鼠神经元形态和超微结构具有保护作用。     

脑积水对脑室周围神经干细胞影响的实验研究

目的探讨脑积水对大鼠脑室周围神经干细胞影响。方法6wWistar大鼠70只，随机分为实验组35只，对照组35只，应用显微外科技术向枕大池内注入25％无菌高岭土混悬液0．05ml，分别在高岭土注射后3d，1w，2w，3w，4w处死动物，迅速取脑组织，测侧脑室指数，用免疫组织化学方法动态检测脑室周围Nestin阳性细胞的表达，同样方法向枕大池内注入生理盐水作为对照组。结果实验组28只大鼠成功诱发脑积水并完成实验全过程，对照组30只完成实验全过程。对照组大鼠各对应时间点，侧脑室指数基本相同，实验组大鼠脑室进行性扩大；脑室周围Nestin阳性细胞：对照组细胞数平均193±20．3个并维持，脑积水造模后3d达到最大值，为对照组308％±1％（P〈0．001），1w脑室周围Nestin阳性细胞下降到对照组196％±1％，2w降到对照组水平210±22．4个，3w脑室周围Nestin阳性细胞几乎看不到并持续。结论脑积水引起脑室进行性扩大，脑室周围神经干细胞可能受脑室扩张机械性压迫引起缺血，缺氧影响，参与脑积水病理损害和修复过程。     

骨髓间充质干细胞移植对脑缺血大鼠突触可塑性影响的实验研究

目的：探讨骨髓间充质干细胞（MSCs）移植对脑缺血大鼠神经功能恢复及突触可塑性的影响。方法：采用大鼠大脑中动脉缺血模型，分为假手术组、模型组、PBS组和MSCs组，研究脑缺血24h后移植MSCs的大鼠神经功能缺损评分（NSS）；分别测定梗死灶周围脑组织突触素（SYN）和脑源性神经营养因子（BDNF）mRNA的表达；电镜及免疫电镜下观察突触结构的变化。结果：与模型组及PBS组大鼠相比，MSCs组的NSS评分较低，SYN及BDNF mRNA的表达则明显较高；电镜检查示MSCs组大鼠突触界面曲率较大，突触后致密物质的厚度增加，突触间隙宽度变窄，突触活性带长度增加；免疫电镜示BrdU阳性细胞和宿主脑神经元形成非成熟的突触样结构。结论：MSCs移植可能通过神经营养效应调节脑缺血周围神经细胞的可塑性改善脑缺血大鼠的神经功能。     

Neuronal injury and brain-derived neurotrophic factor expression in a rat model of amygdala kindling seizures Differences in brain regions and kindling courses

BACKGROUND:Studies have demonstrated that brain-derived neurotrophic factor (BDNF) has a dual effect on epilepsy. However, the relationship between epilepsy-induced brain injury and BDNF remains poorly understood.OBJECTIVE:According to ultrastructural and molecular parameters, to detect the degree of neuronal injury and BDNF expression changes at different brain regions and different kindling times to determine the effects of BDNF on epilepsy-induced brain injury.DESIGN, TIME AND SETTING:A randomized, controlled, animal experiment based on neuropathology and molecular biology was performed at the Department of Physiology and Department of Pathology, Basic Medical College of Jilin University in 2003.MATERIALS:UltraSensitiveTM SP kit for immunohistochemistry (Fuzhou Maxim Biotechnology, China), BDNF antibody (concentrated type, Wuhan Boster Biological Technology, China), JEM-1000SX transmission electron microscopy (JEOL, Japan), and BH-2 light microscope (Olympus, Japan) were used in the present study.METHODS:Wistar rats were randomly assigned to control (n = 6), sham-surgery (n = 6), and model (n = 60) groups. The control group rats were not treated; an electrode was embedded into the amygdala in rats from the sham-surgery and model groups; an amygdala kindling epilepsy model was established in the model group.MAIN OUTCOME MEASURES:Pathological changes in the temporal lobe and hippocampus were observed by light and electron microscopy at 1, 3, 7, 14, and 21 days following kindling, and BDNF expression in the various brain regions was determined by immunohistochemistry.RESULTS:In the model group, temporal lobe cortical and hippocampal neurons were swollen and the nuclei were laterally deviated. There were also some apoptotic neurons 3 days after kindling. The nucleoli disappeared and the nuclei appeared broken or lysed, as well as slight microglia hyperplasia, at 7 days. Electron microscopic observation displayed chromatin aggregation in the nuclei and slight mitochondrion swelling 3 days after kindling. Injury changes were aggravated at 7 days, characterized by broken cytoplasmic membrane and pyknosis. With the development of seizure, the number of BDNF-positive neurons in the hippocampus and temporal lobe increased and peaked at 7 days. Moreover, hippocampal and cortical temporal lobe injury continued. Following termination of electrical stimulation after 7 days of kindling, BDNF expression decreased, but continued to be expressed, up to 21 days of kindling. In addition, the number of temporal and hippocampal BDNF-positive neurons was greater than the control group.CONCLUSION:Brain injury and BDNF expression peaked at 7 days after kindling, and hippocampal changes were significant.     

急性癫痫大鼠海马CA3区Bcl-2表达及线粒体改变

BACKGROUND： Previous studies have shown that the mitochondrial structure and function are damaged in animal models of epilepsy. In addition, the Bcl-2 protein is capable of regulating mitochondrial stability. OBJECTIVE： To observe and validate changes in mitochondrial structure and Bcl-2 expression, and to analyze these characteristics in the hippocampal CA3 region of rat models of epilepsy. DESIGN, TIME AND SETTING： This randomized, controlled, animal experiment was performed at the Laboratory of Electron Microscopy and Department of Histology and Embryology, Luzhou Medical College between 2007 and 2008. MATERIALS： Coriamyrtin was provided by the Pharmacy Factory of West China University of Medical Sciences. The primary and secondary antibodies were provided by Zhongshan Goldenbridge Biotechnology, Beijing. METHODS： A total of 44 adult, male, Sprague Dawley rats were randomly divided into control （n = 11） and epilepsy （n = 33） groups. Rats in the epilepsy group were induced by coriamyrtin （50 μ g/kg）, which was injected into the lateral ventricles. The rats were then observed at 3, 6, and 24 hours after epilepsy induction, with 11 rats at each time point. Epilepsy was not induced in rats from the control group. MAIN OUTCOME MEASURES： Pathological changes in the hippocampal CA3 region were observed by light microscopy; Bcl-2 expression was analyzed by immunohistochemistry; and mitochondrial changes in the hippocampus were observed under transmission electron microscopy. RESULTS： （1） The control group displayed very little Bcl-2 protein expression in the hippocampal CA3 region. However, after 3 hours of epilepsy, expression was visible. By 6 hours, expression peaked and then subsequently decreased after 24 hours, but remained higher than the control group （P 〈 0.05）. （2） Mitochondria were damaged to varying degrees in the epilepsy groups. For example, mitochondria edema, cristae space increase, and disappearance of mitochondria were apparent. Moreover, mitochondrial damage occurred prior to pathological changes in the neurons and nucleolus. CONCLUSION： Bcl-2 expression and mitochondrial damage increased in the hippocampal CA3 region in rats with epilepsy. Moreover, mitochondrial damage occurred prior to increased Bcl-2 expression and nucleolus damage.     

谷氨酸转运体在全脑缺血性癫痫中作用的研究

目的比较三种谷氨酸转运体在全脑缺血性癫痫中的动态变化特征,为癫痫治疗提供有意义靶点.方法SD大鼠以胸部压迫8分钟造成全脑缺血性癫痫模型,分对照组、假手术组、全脑缺血无癫痫组和全脑缺血癫痫组.后两组又根据脑缺血后时间分为6h,24h,48h,72h,5d,7d组.应用免疫组化法研究谷氨酸转运体EAAT-1,EAAT-2,EAAT-3在海马CA1及皮质区表达;研究病理形态变化,同时测定大鼠脑电图改变.结果大鼠癫痫发生率为64%,全脑缺血癫痫大鼠神经损害较无癫痫组严重.与全脑缺血无癫痫大鼠比较,癫痫大鼠海马CA1及皮质区EAAT-2显著持续降低及EAAT-3表达明显升高.结论大鼠癫痫发生与脑缺血严重程度密切相关.海马CA1及皮层区EAAT-2、EAAT-3表达变化是抗癫痫治疗的作用靶点.     

复发缓解型EAE大鼠长期、自然病程的视神经病理改变

目的建立小剂量复合抗原成分诱导的EAE长病程动物模型,通过多种方法观察EAE大鼠的视神经病理改变。方法雌性Wistar大鼠72只,6~8周龄,体重180~200g,其中12只为对照组,余60只诱导EAE模型,进行EAE动物的神经功能评分(按照Kono五级评分法[1]进行),观察大鼠每次发病时的病情、病程变化。于第一次发病后第7d和半年时行眼底照相检查,并取视神经行HE染色光镜观察和透射电镜观察其病理改变。结果 EAE组大鼠视神经光镜主要表现为有不同程度的炎症反应;电镜主要表现为髓鞘稀疏,其髓鞘板层松解,轴突萎缩、变性,结构模糊不清。EAE组第一次发病后第7d及半年时眼底照相提示视乳头萎缩,边界不清楚。半年时视乳头萎缩更明显。结论 EAE大鼠存在明显的视神经病变,主要表现为视神经炎症反应、脱髓鞘改变和轴索损伤。