脑梗死肝阳化风证与阴虚风动证蛋白质组学比较研究

The comparative study on proteomics between cerebral infarction patients with wind-syndrome caused by hyperactive liver-Yang and by Yin deficiency

目的建立脑梗死肝阳化风证与阴虚风动证血清双向凝胶电泳图谱,初步分析其差异蛋白质表达情况。方法双向凝胶电泳(2-DE)分离脑梗死肝阳化风证与阴虚风动证血清标本,考马斯亮兰染色,PD-QUEST图像分析系统分析,从凝胶中选取分离较好的蛋白质点,基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)分析获取肽质量指纹图谱(PMF),Mascot软件搜索SWISS-PROT数据库鉴定蛋白质,对差异蛋白质进行组间对比。结果获得了分辨率较好的脑梗死肝阳化风证与阴虚风动证双向凝胶电泳图谱,质谱分析了29个差异蛋白质点,获取了87张肽质量指纹图谱,通过查询数据库鉴定了29个蛋白质,其中部分蛋白质与肝阳化风证及阴虚风动证发生发展有关。结论建立了脑梗死肝阳化风证与阴虚风动证血清双向凝胶电泳图谱,并应用质谱技术鉴定了部分差异蛋白质点,为脑梗死肝阳上亢证与阴虚风动证与正常组血清双向凝胶电泳图谱血清蛋白质组数据库的建立提供了有意义的数据,并认为蛋白质组学能在脑梗死不同证型具有代表意义。

Objective To establish the serum 2-DE maps of cerebral infarction patients with wind-syndrome caused by hyperactive liver-Yang and by Yin deficiency, and analysize their pro- teomies. Methods Immobilized pH gradient two-dimensional polyacrylamide gel eleetrophoresis （2- DE） was applied to separate the serum proteins of cerebral infarction patients with wind-syn- drome caused by hyperactive liver-Yang and by Yin deficiency. Matrix-assisted laser desorption/ ionization-time of flight mass spectrometry were used to indentify proteins. Results A （MALDI-TOF-MS） and database searching techniques good 2-DE pattern with high resolution was obtained. 29 protein spots were incised from Coomassie brilliant blue staining gel. 87 maps of peptide mass fingerprints （PMF） were obtained and 29 proteins were identified. Some identified proteins were re- lated to cerebral infarction patients with wind-syndrome caused by hyperactive liver-Yang and by Yin deficiency. Conclusion A 2-DE map of the serum of cerebral infarction patients with wind- syndrome caused by hyperactive liver-Yang and by Yin deficiency was established. Some proteins were identified. These data will expand the human serum proteomies database of the normal and the cerebral infarction patients with wind-syndrome caused by hyperactive liver-Yang and by Yin deficiency. The proteomics can delegate cerebral infarction with the different TCM syndromes