脑泰方血清和血浆蛋白质(肽)组学比较研究  

Comparative Study on Naotaifang Containing Serum and Plasma Proteomics(Peptide) by Proteomics

作　　者：王国佐[1] 朱惠斌[2] 杨梅[1] 周瑜[1] 钱荣华[3] 易亚乔[4] 袁紫薇[1] 葛金文[1] 

机构地区：[1]湖南中医药大学中西医结合学院,长沙410208 [2]湖南中医药大学附属第一医院,长沙410208 [3]湖南中医药大学医学院,长沙410208 [4]湖南中医药大学中医学院,长沙410208

出　　处：《世界科学技术-中医药现代化》2013年第7期1505-1514,共10页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金委面上项目(30572370):血清与血浆药理方法对脑泰方药物化学与药效学的对比研究;负责人:葛金文;国家自然科学基金委面上项目(81274008):中药血浆药理方法的建立及系统评价研究;负责人:葛金文;国家自然科学基金委面上项目(81274169):中药复方提取物HNA-1对SIV慢性感染恒河猴幼稚型CD4+T细胞发育;迁移和增殖的影响;负责人:朱惠斌;国家自然基金委青年项目(81202794):特异性蛋白质相互作用网络在脑梗死发生中的作用及中药脑泰方干预机制研究;负责人:王国佐;湖南省教育厅高等学校科学研究项目(12B096):中药脑泰方干预脑梗死分子机制的生物信息学研究;负责人:王国佐;教育部2012年地方高校国家级大学生创新创业训练计划项目(201210541012):脑泰方干预脑梗死机制蛋白质组学研究;主持人:袁紫薇

摘　　要：目的:通过对脑泰方含药血清和含药血浆蛋白质(肽)组学的比较研究,发现脑泰方含药血清和含药血浆中的差异蛋白(肽);以质谱分析技术确定差异蛋白的特征。从而为探索脑泰方的药效物质基础和发展中药血浆药理学方法提供科学依据。方法:取健康成年SD大鼠20只,按体重随机分为空白组、脑泰方给药组,每组10只,连续灌胃给药7天。手术前12 h禁食不禁水,末次给药后1 h,用10%水合氯醛腹腔麻醉,经颈总动脉插管取血。每只大鼠采血后分别制备血清和血浆。每组随机抽取5只大鼠的血清及血浆,采用2-DE技术进行蛋白质(肽)组学比较研究,运用300DPI扫描、PDQuest 7.3.0软件进行分析;采用ESI-MS/MS对重要差异蛋白质点进行鉴定,筛选脑泰方血清和血浆差异蛋白的特征。结果:共获得20张胶图,各胶图分别得到664个蛋白位点。其中与空白血浆组比较,脑泰方血浆样品中共识别到20个差异蛋白点,其中15个差异点表达上调,5个差异点表达下调;与空白血清组比较,脑泰方血清样品中共识别到19个差异蛋白点,其中15个差异点表达上调,4个差异点表达下调;与脑泰方血浆组比较,脑泰方血清样品中共识别到24个差异蛋白点,其中9个差异点表达上调,15个差异点表达下调。选取在脑泰方血浆组和脑泰方血清组中高表达的10个差异蛋白质点进行基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS),成功鉴定了6个蛋白,分别是:inter-alpha trypsin inhibitor,heavy chain 3、Group specific component、complement factor B、Receptor Complexed With A Heterodimeric Fc、complement factor B,isoform CRA-d、Transferrin。结论:脑泰方含药血浆、血清中明显变化的蛋白质可能与其抗凝血促纤溶作用机制相关,这些蛋白参与血管新生、炎症调控等病理生理过程,对研究脑泰方作用的可能有效靶点及其信号转导通路意义重大。Through comparative study on Naotaifang containing serum and plasma proteomics （peptide）, this article revealed differential proteins （peptides） in the Naotaifang. The characteristics of differential proteins were identified with mass spectrometry. It provides scientific evidences for the pharmacodynamic material basis and Chinese herbal medicine plasma pharmacological method development in the exploration of Naotaifang. A total of 20 healthy adult SD rats were randomly divided into the control group, Naotaifang treatment group according to their weights. Ten rats in each group. Intragastric administration of medication was given for seven consecutive days. Before surgery, rats were fed with water but without food. One hour after the last drug administration, 10% chloral hydrate was injected for intraperitoneal anesthesia. Blood was taken through the common carotid artery. Serum and plasma samples were made after blood was taken from each rat. Serum and plasma samples of five rats were randomly selected from each group. And the two-dimensional electrophoresis （2-DE） technique was used in the comparative study of serum pro-teomics （peptide）. The 300 DPI scanning and PDQuest 7.3.0 were used in the analysis. The ESI-MS/MS was used to identify important differences in proteins and screen characteristic serum and plasma protein. The results showed that 20 differential proteins of 5 plasma samples were identified. There were 15 types of proteins expressing up-regulation and 5 types expressing down-regulation. Comparative analysis on the 2-DE gel pictures of Naotaifang containing serum, 19 differential proteins of 5 plasma samples were identified, among which 15 types of proteins express up-regulation and 4 down-regulation. Comparative analysis on the 2-DE gel pictures of Naotaifang containing serum and Naotaifang containing plasma showed that 24 differential proteins of 5 plasma samples were identified, among which 9 types of proteins express up-regulation and 15 down-regulation. The highly expressed pr

关 键 词：脑泰方 血浆 血清 二维电泳 蛋白质组学 质谱 

分 类 号：R392[医药卫生—免疫学]