【作者】 程娜；

【导师】 牛争平；

【作者基本信息】 山西医科大学 ， 神经病学， 2011， 硕士

【摘要】 目的:通过观察wistar大鼠尾静脉注射垂体腺苷酸环化酶激活肽38(PACAP38)后的行为学变化以及三叉神经核尾核(TNC)部位c-fos蛋白的表达情况,探讨PACAP38能否作为诱导剂,诱导大鼠偏头痛发作,以制备新的可靠的实验性偏头痛大鼠模型,并初步探讨PACAP38诱导大鼠偏头痛发作的可能机制。方法:1.选用30只wistar大鼠(250～300g)为研究对象,将其随机分为5组,雌雄各半。空白对照组:大鼠尾静脉注射生理盐水1ml/kg;硝酸甘油模型组:大鼠尾静脉注射硝酸甘油(NTG)2μg/kg;PACAP38低剂量组:大鼠尾静脉给予3μg/kg的PACAP38;PACAP38中剂量组:大鼠尾静脉给予6μg/kg的PACAP38;PACAP38高剂量组:给予12μg/kg的PACAP38。2.尾静脉给予不同实验试剂后即开始观察大鼠行为学变化。主要包括以下内容:①耳红:观察并记录大鼠耳红开始以及结束的时间;②挠头:观察记录大鼠挠头出现及结束的时间,采用持续时间分段计数的方法记录大鼠在尾静脉注射不同实验试剂后每一时间段中挠头的次数;③爬笼:采用持续时间分段计数的方法记录大鼠在尾静脉实验干预后每一时间段内的爬笼次数。3.大鼠尾静脉注射不同实验试剂4h后,腹腔麻醉、灌注固定,于脑立体定位仪上,由延髓闩部向尾端切取4mm脑干标本。应用免疫组织化学技术行c-fos蛋白染色,于光学显微镜下观察c-fos蛋白表达情况。结果:1.行为学比较:①耳红:除空白组外,硝酸甘油组和PACAP38低、中、高三组均出现了耳红,且4组中出现与消失时间相互平行,组间比较无统计学意义(p﹥0.05)。②挠头,硝酸甘油组、PACAP38中剂量组、PACAP38高剂量组出现了挠头次数增多现象,出现及消失时间与耳红结果相似。各分段时间挠头次数统计学比较3组间无显著差异(p﹥0.05),空白组与此3组分别比较有显著差异(p﹤0.05);PACAP38低剂量组较空白组未出现明显的挠头次数增多(p﹥0.05),各分段时间挠头次数与硝酸甘油组比较有统计学意义(p﹤0.05),③爬笼:硝酸甘油组、PACAP38中剂量组、PACAP38高剂量组均较空白组出现了爬笼次数增多现象(p﹤0.05),三组间比较无统计学意义(p﹥0.05);PACAP38低剂量组与空白组比较无明显爬笼次数增多(p﹥0.05),与硝酸甘油组统计学比较有显著性差异(p﹤0.05)。2. c-fos蛋白表达:光学显微镜下可见切片背景为淡黄色或无色,阳性细胞呈黄褐色,多为圆形或卵圆形,以核染为主,易与背景区分,主要分布在三叉神经脊束核尾核(TNC)。硝酸甘油组、PACAP38中剂量组、PACAP38高剂量组大鼠脑干标本c-fos免疫阳性细胞较空白对照组明显增加(p﹤0.05),三组间比较无明显差异(p﹥0.05);PACAP38低剂量组脑干标本c-fos免疫阳性细胞较空白组无明显增加(p﹥0.05),与硝酸甘油组比较有显著性差异(p﹤0.05)。结论:PACAP38可以诱导大鼠偏头痛发作,可能成为新的偏头痛大鼠模型的诱导剂制备偏头痛动物实验模型,并利于更深入的研究偏头痛发病机制及新的防治药物的开发。更多还原

【Abstract】 Objective:Through the observation of the behavioral changes and c-fos expression with trigeminal nucleus tail nuclear (TNC) of the rat following intravenous Pituitary adenylate cyclase activating peptide-38 (PACAP38),to discusses whether PACAP38 can induce migraine attacks in rat to build a new reliable experimental animal model of migraine, and discusses the possible mechanism of PACAP38 induce migraine attacks.Methods:1. 30 experimental migraine rat weighted 200~250g were randomly divided into five groups: sham group(group1),NTG group(group2), PACAP38 LD group (group3), PACAP38 MD group(group4),PACAP38 HD group(group5),1ml/kg physiological saline infusion to sham group;2μg/kg glyceryltrinitrate infusion to NTG group;3μg/kg PACAP38 infusion to PACAP38 LD group;6μg/kg PACAP38 infusion to PACAP38 HD group;12μg/kg PACAP38 infusion to PACAP38 HD group.2. To appraise behavior changes of the rat.The time of turning red of ears and frequency of scratching head and climbing hutch were observed every 30 minuts after infusion and comparae with eachother.3.4h after infusion,paralyzerat,rat were paralyzed and perfuse to fix,which the brainstem were removed and post-fixed.The expression of c-fos immuno-reactivities was detected with immunohistohistochemical method.Results:1.Ethology comparison:①red ears: beside group1,group2,3,4,5 were appeared reddish two ears. There is no statistical difference in the time of turning red of ears in group2,3,4,5.(p﹥0.05).②scratching head:group2,4,5 appeared a phenomenon of scratching head.There is no statistical difference in the frequency of scratching head in group2,4,5.Compared with group1,the frequency of scratching head was significantly increased(p﹤0.05). There is no statistical difference in the frequency of scratching head between group1,3,and compared with group2 , the frequency of scratching head was significantly lower(p﹤0.05).③climbing hutch: group2,4,5 appeared a phenomenon of climbing hutch.There is no statistical difference in the frequency of climbing hutch in group2,4,5.Compared with group1,the frequency of climbing hutch was significantly increased(p﹤0.05). There is no statistical difference in the frequency of climbing hutch between group1,3.and compared with group2 ,the frequency of climbing hutch was significantly lower(p﹤0.05).2. In microscope,the cell was tasteless in background, the positive cell was yellow in TNC. The positive cell in group2,4,5 was obviously increased comrared with group1(p﹤0.05),and no signigicant difference between eachother(p﹥0.05). Compared with group1,there is no obviously increased in group3(p﹥0.05).Conclusion:PACAP38 can induce rats migraine attacks,and can be used as inducer to build a new reliable experimental animal model that can mimic the migraine attacks,and can be used in the basic research on the mechanism,therapeuical drug and other approaches of migraine.更多还原