【作者】 李智；

【导师】 陈泽奇；

【作者基本信息】 中南大学 ， 中西医结合临床， 2007， 硕士

【摘要】 目的：采用蛋白质组学技术研究实验性偏头痛肝阳上亢证大鼠与正常大鼠蛋白质组表达的差异，探寻偏头痛肝阳上亢证的相关蛋白。方法：将20只SD大鼠随机分成正常组及偏头痛肝阳上亢证组(简称模型组)，每组各10只。肝阳上亢证动物模型的复制参照金益强的方法，偏头痛模型复制采用Cristina Tassorelli硝酸甘油法。造模成功后分别取正常组及模型组大鼠下丘脑和肾上腺组织为研究标本，并对组织蛋白质进行提取，每组标本经3次双向电泳，PDQuest软件分析，以及差异蛋白质点的质谱分析和鉴定。结果：1．发现模型组与正常组大鼠下丘脑2-DE的总蛋白质分布模式非常相似，以pI4～8，M_r20～75kD范围的蛋白质斑点分布最多。经PDQuest 7.0软件分析后得出，在相同条件下识别的蛋白质斑点数分别为模型组556±23个，正常组482±20个。结合肉眼可以明显区分13个表达上调的蛋白质斑点．将13个蛋白质斑点进行质谱分析和鉴定，其中有6个点的蛋白质得到有效鉴定。2．同样发现模型组与正常组大鼠肾上腺总蛋白质分布以PI3—8，Mr14.4—75kD范围最多。正常组有584±25个蛋白质点，模型组有686±27个蛋白质点。可以明显区分的12个表达上调的蛋白斑点中有9个点的蛋白质得到有效鉴定结论：1．本研究初步建立了实验性偏头痛肝阳上亢证下丘脑和肾上腺组织的固相PH梯度双向凝胶电泳图谱。2．通过凝胶图谱分析，模型组与正常组大鼠下丘脑和肾上腺组织2—DE图谱的表达具有差异性。3．通过差异蛋白质点的质谱分析，鉴定出15个蛋白质点。推测其中的乙醛脱氢酶、谷胱甘肽、蛋白质二硫化物异构酶前体可能为肝阳上亢证的相关蛋白。β微管蛋白、α微管蛋白、β瘢痕蛋白以及突触[小体]膜联合25K蛋白可能为偏头痛的相关蛋白。4．这为以后进一步明确偏头痛肝阳上亢证的发病机制打下了基础，亦为研究其诊断、治疗提供了新的思路。更多还原

【Abstract】 AIM: This experiment aims to analyze the differentially expressed proteins between experiment migraine rat with hyperactivity of liver-yang and the normal rat by proteome technology. Explore the proteins associated with the migraine rat with hyperactivity of liver-yang.METHODS: Divide the 20 SD rats to the normal group and the migraine with hyperactivity of liver-yang group (the model group) by random. Every group has 10 rats. The copy of the rat with hyperactivity of liver-yang according to the method of Jin Yi-qiang. Apply Cristina Tassorelli nitroglycerin method to copy the migraine model. After successfully making the model, taking hypothalamus and adrenal glands tissues of the normal group and the model group by each. Every group tissue proteins were extracted and further analyzed by two-dimensional gel electrophoresis for three times and PDQuest 7.0 software. Then select the different protein spots for mass spectrometry and identification.RESULT: 1. The total protein distribution pattern of the model group and the normal group rat hypothalamus 2-DE is very similar. Protein spots were mainly displayed at pI4-8, Mr20-75kD range. Through PDQuest 7.0 software analyzing , there were 556±23 protein spots in the mode group while 482±20 spots in the normal group. Take the 13 high expressed protein spots that notable expressed to mass spectrographic analysis and identification. There are 6 protein spots were utility identified.2. The total protein distribution pattern of the model group and the normal group rat adrenal glands protein spots were mainly displayed at pI3-8, Mr14.4-75kD range, there were 686±27 protein spots in the mode group while 584±25 spots in the normal group. There are 9 protein spots were utility identified in the 12 high expressed protein spots that notable expressed.CONCLUSION: 1.This study established rudiment immobilized PH gradient-based SDS-PAGE two-dimentional gel electrophoresis image maps of hypothalamus and adrenal glands of the experiment migraine rat with hyperactivity of liver-yang. 2.Via 2-DE profile analyzing, there are differences in the model group and the normal group of hypothalamus and adrenal glands proteins3.Via mass spectrometry of different proteins, identifying 15 protein spots. Infer the aldehyde dehydrogenase glutathione peroxidase and protein disulfide-isomerase precursor mabe concerned with the hyperactivity of liver-yang. The Beta tubulin alpha tubulin Beta-synuclein and synaptosomal associated 25K protein mabe concerned with the migraine.4.The result to do the groundwork for identifying the morbidity mechanism. This also provide new idea for studying the diagnosis and the treatment.更多还原