基于小型猪冠心病心肌缺血模型的血瘀证蛋白组学研究  

Proteomics of blood stasis syndrome based on myocardial ischemia model of coronary heart disease in minswines

作　　者：王勇[1] 啜文静[1] 郭淑贞[1] 李春[1] 王伟[1] 

机构地区：[1]北京中医药大学基础医学院,北京100029

出　　处：《北京中医药大学学报》2011年第7期460-464,共5页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家科技重大专项资助项目(No.2009ZX09502-018);国家自然科学基金资助项目(No.30902020)

摘　　要：目的寻找冠心病血瘀证的特征差异蛋白,探索血瘀证可能的内在病理机制。方法采用中华小型猪左冠状动脉前降支起始部远端放置Ameroid缩窄环的方法制备慢性心肌缺血血瘀证模型。小型猪随机分为假手术组和模型组。动态观察模型动物心肌缺血(0～ 8周)情况,冠脉造影观测冠脉狭窄程度;采用双向凝胶电泳和基质辅助激光解析/电离-飞行时间质谱(MALD I-TOF)对10只假手术组、9只模型组动物进行比较蛋白质组学研究,实验后期利用W estern blot法对相关差异蛋白进行初步验证,以进一步确定冠心病心肌缺血血瘀证的差异蛋白。结果初步研究发现心肌缺血血瘀证模型组动物心肌蛋白组学研究中,模型组动物与假手术组相比,有5个明显的下调蛋白点、8个明显的上调蛋白点,对其中部分差异蛋白点进行了MALD I-TOF质谱鉴定,结果同时得到了二级质谱的确定。差异蛋白主要是氧化应激蛋白及相关的心肌结构蛋白,包括热休克蛋白27(HSP27)、过氧化物酶(peroxiredoxin 3 isoform 1),心肌肌钙蛋白T(cTnT)、肌球蛋白(myosin lightpolypeptide)。结论冠心病慢性心肌缺血血瘀证在心肌局部表现为氧化应激损伤以及心肌结构蛋白的改变,相关的差异蛋白可作为临床冠心病心肌缺血血瘀证药物治疗的靶标。Objective To find the differential proteins of blood stasis syndrome of coronary heart disease （ CHD）, and explore the possible internal pathogenesis of blood stasis syndrome. Methods The model of blood stasis syndrome of chronic myocardial ischemia was established by applying the method of placing Ameroid ring in the left anterior descending coronary artery in Chinese miniswines. All miniswines were randomly divided into the sham-operation group and model group. The status of myocardial ischemia （0 -8 weeks） was observed dynastically in the model. The stenosis of coronary artery was observed by using coronary angiography （CAG）. The study of comparative proteomics was conducted by applying two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry （MALDI-TOF-MS） in 10 miniswines of the sham-operation group and 9 ones of the model group. The relevant differential proteins were verified initially by using Western blot method at later stage of the test for determining further the differential proteins of CHD myocardial ischemia. Results The initial study found out that there were 5 distinct down-regulated protein spots and 8 distinct up-regulated protein spots in the model group compared with the sham-operation group. The identification of MALDI-TOF-MS was used in haass spectrum at the same some differential protein spots and the result was determined by second-order time. The differential proteins were mainly oxidative stress proteins and relevant myocardial structural proteins, including heat shock 27 kDa protein （HSP27）, peroxiredoxin 3 isoform 1, cardiac troponin T （cTnT） and myosin light polypeptide. Conclusion The myocardial manifestations were observed as oxidative stress lesion and changes of myocardial structural proteins in blood stasis syndrome of CHD chronic myocardial ischemia. The relevant differential proteins can be taken as the target of medicinal treatment.

关 键 词：冠心病 慢性心肌缺血 血瘀证 蛋白组学 小型猪 

分 类 号：R541.4[医药卫生—心血管疾病]