大鼠急性肺栓塞血清差异表达蛋白的研究

Comparative proteome analysis of serum in rat model of acute pulmonary embolism

目的研究急性肺栓塞(APE)大鼠血清和正常大鼠血清的差异蛋白表达变化。方法采用血栓颈静脉注入法制备大鼠APE模型,采用双向电泳技术找出差异蛋白,用MALDI-TOF技术和生物信息学技术鉴定差异蛋白,并对部分差异表达蛋白采用Western-blot技术作进一步验证。结果大鼠血清蛋白的2-DE胶银染可分离出1 400多个点,考染可达到1 200个点以上。经图像分析得到的24个差异表达蛋白中有12个蛋白得到鉴定。对部分差异蛋白采用Western-blot方法在蛋白水平的验证结果与2-DE结果基本相符。结论采用比较蛋白质组学的方法可以发现APE大鼠血清中的差异表达蛋白,这些差异表达蛋白分子将为我们寻找新的APE血清学标志物提供重要的线索和依据。

AIM To find differentially expressed serum proteins in rat acute pulmonary embolism （APE） model. METHODS We constructed a rat APE model by injecting 3 -4 emboli into the left jugular vein. The blood samples were collected by intracardiac puncture at different time points and the serum proteins were separated by two-dimensional electrophoresis （2-DE）. The differentially expressed proteins were identified using matrix assisted laser desorbtion/ionization time-of-flight mass spectrometer （MALDI-TOF-MS）. Some of the identified proteins were validated by western blot analysis. RESULTS There were approximately 1 400 protein spots in every 2-DE gels of serum protein samples visualized by silver staining and 1 200 protein spots for Coomassie Brilliant Blue R-250. Of the twenty-four differentially displayed protein spots, we found corresponding protein candidates in the database for twelve of them. The results of western blot analysis were in accordance with those of 2-DE. CONCLUSION The differential expression patterns of these proteins suggest the distinct roles they may play in different stages of the rat APE model and findings from this study may lead us to find some new useful biomarkers in the diagnosis of APE.