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Batroxobin plus hypothermia for protection of cerebral ischemia/reperfusion injury models in gerbils

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Neural Regeneration Research 2006年第5期1卷 405-407页

作者： Lin Zhang Pixing Zhang Yinming Zeng Qun Chen Department of Anesthesiology Affiliated Hospital of Qingdao University Medicat CollegeQingdao 266003 Shandong Province China First Clinical College China Medical University Shenyang 110001 Liaoning Province China Department of Anesthesiology Eighth People＇s Hospital of Qingdao City Qingdao 266000 Shandong Province China Jiangsu Key Lab of Anesthesiology Xuzhou 221002 Jiangsu Province China

BACKGROUND： Hypothermia plays a protective role in cerebral ischemialreperfusion injury. Dose combination with batroxobin, an active drug for treating cerebrovascular disease, will enhance its protection？ OBJECTIVE ： To explore the effects of hypothermia, batroxobin, hypotherrnia combined with batroxobin on complete cerebral ischemia/reperfusion injury in gerbils. DESIGN ： A randomized block comparison observation.SETTING ： Jiangsu Key Lab of Anesthesiology. MATERIALS ： Experimental animal： Sixty Mongolia gerbils weighing 50-80 g, male or female, were provided by the Animals Center of Xuzhou Medical College. Drugs and agents： Batroxobin was provided by Dongling Pharmaceutical Industry Organization （Japan）. Superoxide dismutase （SOD） and malondiadehyde （MDA） kits were offered by Nanjing Jiancheng Bioengineedng Institute. Other reagents were all import or national analytical pure grade. HITACHI R22A refrigerated high-speed centrifuge, and HARRIS ultra-hypotherrnia refrigerator were used. METHODS ： The experiments were completed in Jiangsu Key Lab of Anesthesiology from May 2004 to January 2005. ① The animals were divided into 6 groups by random member table method： sham-operated group （n =6）, ischemia control group （n =6）, norrnothermia group （n =12）, hypothermia group （n =12）, batroxobin group （n =12） and hypothermia＋batroxobin group （n =12）. Gerbil rats were abdominally anesthetized with sodium pentobarbital. The neck skin was incised to separate bilateral common carotid arteries. Complete cerebral ischemia models were established by occluding bilateral common carotid arteries with artery clamp for 10 minutes, then the clamp was loosened to perfuse the arteries, Iso-electric level of brain electric wave showed the models were established successfully. The gerbils in the batroxobin group and hypotherrnia＋batroxobin group were abdominally injected with batroxobin （8 BU/kg） while reperfusion, and isovolumetric saline was administered to the gerbils in the oth

关键词：巴曲酶/药理学丙二醛/代谢超氧化物歧化酶/代谢低温疾病模型动物脑缺血/病理生理学脑缺血/病理学沙鼠亚科再灌注损伤/代谢大鼠动物男(雄)性女(雌)性

西洛他唑鼻腔给药对小鼠慢性脑缺血损伤的保护作用

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浙江大学学报（医学版） 2011年第2期40卷 169-175页

作者：张琦叶夷露李青张纬萍姜莉莉魏尔清俞月萍浙江医学高等专科学校药理教研室浙江杭州310053 浙江大学医学院药理学系浙江杭州310058 温州医学院药学院浙江温州325035

目的:观察西洛他唑鼻腔给药对小鼠慢性脑缺血损伤的保护作用。方法:以大脑中动脉栓塞(MCAO)诱导小鼠局灶性脑缺血,分别于术后1、4、7 h给予西洛他唑(鼻腔给药或腹腔注射给药),从第2天起,每天2次,持续到缺血后14 d。缺血后1、3、7、14、2... 详细信息

目的:观察西洛他唑鼻腔给药对小鼠慢性脑缺血损伤的保护作用。方法:以大脑中动脉栓塞(MCAO)诱导小鼠局灶性脑缺血,分别于术后1、4、7 h给予西洛他唑(鼻腔给药或腹腔注射给药),从第2天起,每天2次,持续到缺血后14 d。缺血后1、3、7、14、21、28、35 d进行神经症状评分和斜板试验,并记录小鼠的生存率。实验结束后,测定脑损伤体积和神经元数量。结果:西洛他唑鼻腔给药0.3 mg/kg能显著提高小鼠MCAO术后35 d的生存率。西洛他唑鼻腔给药(0.3、1mg/kg)和西洛他唑腹腔给药(10 mg/kg)能改善缺血后神经症状评分,降低脑梗死体积,减轻脑萎缩,并不同程度地增加缺血侧不同脑区的神经元密度。结论:西洛他唑鼻腔给药对小鼠慢性缺血性脑损伤具有保护作用,可作为脑缺血治疗的一种新方法。

关键词：脑缺血/药物疗法脑缺血/病理学四唑类/治疗应用疾病模型,动物西洛他唑鼻腔给药

Characteristics of global cerebral ischemia models constructed by modified four-vessel occlusion in rats

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Neural Regeneration Research 2006年第5期1卷 436-440页

作者： Jinbao Li Lai Jiang Hua Xu Yuanchang Xiong Xiaoming Deng Department of Anesthesiology Changhai Hospital Second Military Medical University of ChinesePLA Shanghai 200433 China

BACKGROUND： Pulsinelli et al developed a kind of rat models with four-vessel occlusion-induced global cerebral ischemia, Because the histo-pathological changes and severe cerebral ischemia reproducibility of this model are good and the stability of this model in circulation respiration is superior to that of other models, so four-vessel occlusion method has become a classic modeling method for global cerebral ischemia model, This model has been improved in some laboratories to meet different requirements in different studies. OBJECTIVE： To establish a highly reproducible rat model of reversible forebrain ischemia by modifying four-vessel occlusion model introduced by Pulsinelli et al, and to investigate its neurophysiological and pathological changes and the characteristics of modified operation. DESIGN ： Completely randomized grouping, controlled tria. SETTING ： Department of Anesthesiology,Changhai Hospital, Second Military Medical University of Chinese PLA. MATERIALS ： A total of 65 male healthy SD rats, weighing 250-300 g, were provided by the Experimental Animal Center of Second Military Medical University of Chinese PLA. VSM hemodynamic monitor and temperature monitor （Thermal ert TH-5, U.S.A） were used. METHODS ： The trial was conducted in the Department of Anesthesiology, Changhai Hospital, Second Military Medical University of Chinese PLA from January 2005 to March 2006. ① Experimental grouping： Sixty-five rats were randomly divided into the following 7 groups： sham-operation group （n =9）： given the same operation, without occlusion of vessels; 5 minutes ischemia group （n =9）： ischemia 5 minutes and reperfusion 72 hours; 10 minutes ischemia group （n =8）： ischemia 10 minutes and reperfusion 72 hours; 15 minutes ischemia group （n =9）： ischemia 15 minutes and reperfusion 72 hours; 20 minutes ischemia group （n =8）： ischemia 20 minutes and reperfusion 72 hours; 30 minutes ischemia group （n =7）; ischemia 30 minutes and reperfusion 72 hours; ischemia con

关键词：疾病模型动物脑缺血/病理生理学脑缺血/病理学血压再灌注损伤/病理生理学大鼠动物男(雄)性

Changes of cerebral blood flow in rats with acute cerebral ischemia and the effect of nitric oxide donor S-nitroso-N-acetyl-penicillamine

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Neural Regeneration Research 2006年第5期1卷 393-396页

作者： Feng Gao Zhiqiang Yi Guijun Lin Department of Neurosurgery Aerospace Central HospitalPeking University Beijing 100049 China

BACKGROUND ： Previous studies show that nitric oxide donor can increase cerebral blood flow and improve the function of neurons in cerebral ischemia, but the change does not happen in all the models of cerebral ischemia. OBJECTIVE ： To observe the effects of nitric oxide donor S-nitroso-N-acetyl-penicillamine （SNAP） on the cerebral blood flow, cyclic guanosine monophosphate （cGMP） content in cerebral cortex, infarct volume and blood pressure in acute ischemic rat brain.DESIGN ： A randomized and control animal experiment SEI-FING ： Department of Neurosurgery, Aerospace Central Hospital, Peking University.MATERIALS： Twenty-eight male Wistar rats of SPF grade, weighing 250-300 g, aged 10-12 weeks were randomly divided into control group （n =14） and SNAP-treated group （n =14）. SNAP （5 mg/bottle） was provided by Beijing Chemical Reagent Company. Laser Doppler Flowmeter （FLO C1; Omegawave Inc., Tokyo, Japan） and immunoassay kit （Amersham Pharmacia Biotech, UK） were applied. METHODS： ① Model establishment： In the control group, models of cerebral ischemia were induced by ligating right common, internal and external carotid arteries; In the SNAP-treated group, models of cerebral ischemia were induced by ligating right common and external carotid arteries, followed by occluding middle cerebral artery and ligating internal carotid artery. ②Administration： In the SNAP-treated group, SNAP （100 μg/kg） was intravenously infused within 2 minutes, whereas in the control group, phosphate buffered saline （PBS, 1 mL） was intravenously infused （0.5 mL per minute）. Six rats were used to measure the volume of cerebral infarction, and the other 8 rats were used to determine other indexes in each group respectively. ③ Determination of indexes： Regional cerebral blood flow （rCBF） was continuously measured by laser-Doppler flowmetry in the ischemic penumbra and contralateral cortex under the continuous monitoring of blood pressure, cGMP concentrations in brain tissue were determined

关键词：钙调蛋白结合蛋白质类/代谢环GMP/代谢疾病模型动物脑缺血/代谢脑缺血/病理学青霉胺/药理学一氧化氮/代谢大鼠动物老年人男(雄)性

Effects of hypoxic preconditioning on the changes of expression of neuroglobin mRNA and labeled positive cells following cerebral ischemia in gerbils

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Neural Regeneration Research 2006年第5期1卷 397-400页

作者： Yong Zhang Yanqun Chang Zhenfang Liu Qingxi Fu Xiao Zhang Department of Neurology Shandong Provincial HospitalShandong University Jinan250021 Shandong ProvinceChina Depayment of Anesthesiology Shandong Provincial HospitalShandong University Jinan250021 Shandong ProvinceChina

BACKGROUND： Neuroglobin （NGB）, as newly discovered the third member of the globin family binding oxygen, mainly exists in brain of human beings and vertebrates, and it is closely correlated with the oxygen supply in brain. OBJECTIVE： To observe the changes of the expression of neuroglobin and number of positive cells labeled immunohistochemically following cerebral ischemia in gerbils after hypoxic preconditioning. DESIGN ： A complete randomized grouping design and controlled experiment.SETTING ： Department of Neurology, Department of Anesthesia, Shandong Provincial Hospital, Shandong University. MATERIALS ： Sixty-six adult male Mongolian gerbils of clean degree, about 50-65 g, at an average of 57.5 g were provided by the Experimental Animal Center of Capital Medical University [certificate number of animal quality：SCXK（Beijing）2000-0012]. TRNzil （Tianwei Shidai Company, Beijing）, polymerase chain reaction （PCR） primers （synthetized by Invitrogen Company, Shanghai）; reverse transcription-PCR （RT-PCR） one-step kit （Toyobo Company）; PCR instrument （GeneAmp PCR System 240）; mice brain NGB monoclonal antibody （Academy of Military Medical Sciences）; DAB （Zhongshan Company, Beijing）. METHEDS ： The study was completed from December 2004 to June 2005 in Shandong Provincial Hospital. ① The 66 gerbils were randomly divided into sham-operated group （n =6）, cerebral ischemia group （n =30） and hypoxic preconditioning group （n =30）. The gerbils in the hypoxic preconditioning group were put in the environment which contained 02 （0.08 in volume fraction） and N2 （0.92 in volume fraction） at temperature of 25 ℃ for 2 hours. After 5 hours, the gerbils in the hypoxic preconditioning group and cerebral ischemia group were anesthetized, then bilateral common carotid arteries were ligated. In the sham-operated group, bilateral common carotid arteries were only isolated without ligation and hypoxic preconditioning. ② At 1, 5, 10, 30 and 60 minutes after cerebral ischemia, the exp

关键词： RNA 信使/遗传学疾病模型动物脑缺血/代谢脑缺血/病理学缺血预处理沙鼠亚科一氧化氮/代谢成年人动物男(雄)性人类小鼠

全脑缺血再灌注小鼠额叶和海马c-fos基因的表达

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中国临床康复 2006年第26期10卷 63-65,i0001页

作者：王红艳李威孙博谦姜立刚李海平北华大学附属医院神经科吉林省吉林市132011

目的:探讨c-fos基因表达在全脑缺血再灌注后不同时间的变化规律。方法:实验动物全部采用由白求恩医科大学实验动物中心提供的雄性昆明小鼠,6～7周龄,体质量25～28g。①动物分组:第1批:40只小鼠随机分为假手术组5只,脑缺血再灌注组35只,... 详细信息

目的:探讨c-fos基因表达在全脑缺血再灌注后不同时间的变化规律。方法:实验动物全部采用由白求恩医科大学实验动物中心提供的雄性昆明小鼠,6～7周龄,体质量25～28g。①动物分组:第1批:40只小鼠随机分为假手术组5只,脑缺血再灌注组35只,根据再灌注不同时间提取标本,分别为术后1h,1d,3d,7d,2周,4周,6周,每个时间点5只。第2批:40只小鼠用于反转录-聚合酶链反应,分组同上。②模型制备及检测方法:制备全脑缺血再灌注损伤模型,假手术组不阻断颈总动脉亦不放血。采用卒中指数对各组小鼠神经病学症状进行评价。于小鼠双侧颈总动脉阻断再灌注后1h,1d,3d,7d,2周,4周,6周取小鼠额叶皮质、海马区脑组织采用免疫组织化学染色、反转录-聚合酶链反应技术,对c-fos基因表达变化进行检测。结果:80只小鼠均进入结果分析,无脱落。①免疫组化检测c-fos基因表达产物Fos蛋白结果:假手术组Fos蛋白有较弱表达,免疫组化染色假手术组未见Fos蛋白阳性神经元;再灌注1h在额叶皮质与海马区即可见Fos蛋白表达升高,与假手术组差异显著,再灌注1dFos蛋白表达达最高峰,并持续至2周(P脑缺血再灌注可诱导中枢神经系统c-fos基因的持续表达。

关键词：脑缺血/病理学再灌注基因,fos 原癌基因蛋白质c-fos/血液

高压氧对大鼠脑缺血区线粒体氧自由基影响的实验研究

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浙江大学学报（医学版） 2008年第5期37卷 437-443页

作者：楼敏王季华钱琼秋闻树群丁美萍浙江大学医学院附属第二医院神经内科浙江杭州310009 浙江大学园艺系浙江杭州310029

目的:研究高压氧(HBO)对大鼠脑缺血区线粒体氧自由基及抗自由基酶的影响。方法:参照改良的Koizumi方法,在激光多普勒血流仪监测局部脑血流的条件下,建立线栓法大鼠局灶性脑缺血模型,缺血90min后再灌注,缺血后3h高压氧治疗(3个标准大气压... 详细信息

目的:研究高压氧(HBO)对大鼠脑缺血区线粒体氧自由基及抗自由基酶的影响。方法:参照改良的Koizumi方法,在激光多普勒血流仪监测局部脑血流的条件下,建立线栓法大鼠局灶性脑缺血模型,缺血90min后再灌注,缺血后3h高压氧治疗(3个标准大气压,1h),缺血后24h分别取缺血核心区和半暗区脑组织,差速离心提取线粒体,进行超氧阴离子自由基(O2.)生成速率、H2O2含量、丙二醛(MDA)含量测定,以及线粒体超氧化歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)和还原型谷胱甘肽(GSH)含量测定。结果:脑缺血再灌注24h后缺血半暗区和核心区线粒体的H2O2、O2.、MDA的含量较正常对照组明显增加,而SOD、GSH-PX活性以及GSH的含量较正常对照组明显下降(P缺血半暗区线粒体O2.含量增加(P缺血核心区线粒体O2.含量增加(P脑缺血区的H2O2、GSH-PX和GSH作用无影响。结论:在脑缺血时间窗内,HBO治疗能增加脑缺血区线粒体自由基生成,提高线粒体抗自由基酶活性;能抑制脑缺血半暗区线粒体的脂质过氧化损伤,但对核心区作用不明显。提示线粒体的功能状态在HBO治疗后的自由基反应中起重要作用。

关键词：高压氧脑缺血/治疗脑缺血/病理学线粒体活性氧/代谢再灌注损伤/病理学再灌注损伤/治疗疾病模型动物

神经节苷脂对脑缺血再灌注大鼠突触素p38表达的影响

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中国临床康复 2006年第22期10卷 87-89页

作者：高永哲杨四清杨敏武汉大学医学院中南医院神经科武汉大学医学院脑血管病研究中心湖北省武汉市430071

目的:观察神经节苷脂对大鼠脑缺血再灌注后突触素p-38表达的影响。方法:成年健康雌性Wistar大鼠54只,随机分为神经节苷脂治疗组、对照组和假手术组,每组18只。采用线栓法建立大脑中动脉缺血再灌注动物模型,假手术组不插入尼龙线。神经... 详细信息

目的:观察神经节苷脂对大鼠脑缺血再灌注后突触素p-38表达的影响。方法:成年健康雌性Wistar大鼠54只,随机分为神经节苷脂治疗组、对照组和假手术组,每组18只。采用线栓法建立大脑中动脉缺血再灌注动物模型,假手术组不插入尼龙线。神经节苷脂治疗组大鼠在大脑中动脉缺血再灌注手术前2h和术后每天脑室注神经节苷脂10g/L,对照组同步脑室注射等量10μL生理盐水,两组分别于治疗后6h,1,3,7,14,21d各取3只,40g/L多聚甲醛心脏灌注固定,取脑备用。假手术组不作处理,于相应时间点心脏灌注固定,取脑备用。应用免疫组织化学方法观察神经节苷脂对p-38表达的影响。结果:54只大鼠均进入结果分析。①大体观察:在大脑中动脉缺血再灌注24h后左侧大脑中动脉供血区脑组织肿胀、苍白,第7天可见顶叶颗粒状局部脑组织萎缩,第14天有坏死囊腔形成。②p38表达:假手术组在皮质、纹状体和海马区p38表达很弱,左右对称,其校正吸光度值在不同时间点比较,差异无显著性意义。对照组纹状体背外侧梗塞周围区,p38的免疫活性于再灌注6h和1d未出现明显变化,第3天可见升高,第7天达到高峰,第14天明显下降,第21天下降至假手术组水平。在缺血侧顶叶皮质亦可见到p38的免疫活性在3d后明显升高,在21d已接近假手术组水平。脑缺血再灌注3～7d,p38免疫活性最高,免疫产物的颗粒较大,其校正吸光度值显著高于假手术组(P<0.05)。脑室注射外源性神经节苷脂组,各脑区p38的表达较对照组略有增强,各个时间点的校正吸光度值与对照组比较差异无显著性意义。结论:脑室注射外源性神经节苷脂后,各脑区P38免疫反应活性有所增加,但差异不显著,不能得出神经节苷脂与P38的直接因果关系,亦不能断然否定两者间的内在联系。

关键词：神经节苷脂类脑缺血/病理学大鼠

Electroacupuncture at Ren channel point combined with injection of basic fibroblast growth factors for neural stem cell proliferation in subventricular zone of cerebral ischemic side

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Neural Regeneration Research 2006年第5期1卷 424-427页

作者： Min Pi Lihong Diao Haibo Yu Xiaodan Rao Ling Wang Yuansheng Liu Wenshu Luo Zhuoxin Yang Department of Acupuncture and Moxibustion Shenzhen Hospital Guangzhou University of Traditional Chinese Medicine Shenzhen 518033 Guangdong Province China Postdoctoral Station of Guangzhou University of Traditional Chinese Medicine Guangzhou 510405 Guangdong Province China

BACKGROUND： Electroacupuncture at Ren channel point can treat stroke and generate some neurodevelopment-related growth factors, which are associated with the proliferation and differentiation of stem cells. OBJECTIVE： To observe the effect of electroacupuncture at Ren channel point combined with injection of basic fibroblast growth factors （bFGF） on the expression of 5-bromo-2-deoxyuridine （BrdU）-Iabeled and BrdU/Neuroepthelial stem cell protein （Nestin）-Iabeled positive cells in subventricular zone of cerebral ischemic rats. DESIGN ： Completely randomized grouping design and controlled trial.SETTING： Department of Acupuncture and Moxibustion, Shenzhen Hospital, Guangzhou University of Traditional Chinese Medicine. MATERIALS ： Eighty-four adult healthy male Wistar rats were involved and divided into 5 groups by a lot： blank group （n =6）, sham-operation group （n =6）, model group （n =26）, injection of bFGF group （bFGF group, n =24） and electroacupuncture at Ren channel combined with injection of bFGF group （bFGF group, n =22）. Three time points were set in each of the latter three groups for observation： 7, 14 and 28 days after ischemia respectively. Mouse BrdU and Nestin monoclonal antibody （analytical pure, China）, G6805 electro-acupuncture apparatus （Huayi Medical Instrument, Shanghai） and Olympus FV 500 laser confocal microscope （Olympus Company） were used. METHODS ： This trial was carried out in the Laboratory of Anatomy, Medical College of Sun Yat-sen University from September 2004 to July 2005. ① Except for blank group and sham-operation group, the rats in the other groups were prepared into rat models of local cerebral ischemia by suture-occluded method. The rats were reperfused, and then they were graded in neurologic deficits based on neurological scoring of Longa. Rat model was successful when the neurological scoring ranged from 1 to 4 points and used for the trial. Blank group was untouched; Right common carotid artery was isolated and right exter

关键词：成纤维细胞生长因子2/代谢电针疾病模型动物脑缺血/代谢脑缺血/病理学神经组织蛋白质类/代谢细胞增殖/针灸效应中间丝蛋白质类/代谢成年人大鼠动物男(雄)性小鼠