基于双向凝胶电泳技术的中医寒、热体大鼠蛋白质组表达谱研究

Research on Proteomic Spectra Expression of Traditional Chinese Medicine Cold and Heat Constitution Rats Based on Two-dimensional Gel Electrophoresis

目的:利用双向凝胶电泳技术对中医寒、热体大鼠进行蛋白质组表达谱研究,以期寻找与寒热体相关的蛋白质,并探讨中医寒、热体形成的生物学基础。方法:提取大鼠肝脏细胞的总蛋白,采用双向凝胶电泳(2-DE)及MALDI-TOF-MS质谱技术筛选并鉴定寒热体表达差异的蛋白质群。结果:经过筛选和质谱技术共获得10个具有统计学意义的蛋白表达差异点:氨甲酰磷酸合成酶、二硫键异构酶、过氧化氢酶、过氧化氢异构酶、细胞质氨肽酶、谷氨酸脱氢酶、3-羟基-3-甲基戊二酰辅酶、热休克蛋白前体、同型半胱氨酸、葡萄糖调节蛋白前体。结论:中医寒热体大鼠在蛋白质组表达方面存在一定的差异性,酶类蛋白代谢异常可能是寒热体形成的物质基础之一。

This article was aimed to study the proteomic spectra expression of traditional Chinese medicine（TCM） cold and heat constitution rats with two-dimensional gel electrophoresis（2-DE）, in order to search for the cold and heat-associated proteins for the investigation of the biological basis of TCM cold and heat body constitution formation. The total protein in rat＇s liver cell was extracted. The 2-DE and MALDI-TOF/TOF mass spectrometry（MS） were used in the screening and identification of differentially expressed proteins of cold and heat constitution rats. The results showed that a total of 10 different points in the protein expression were obtained with statistical significance after screening and MS, which were carbamoyl-phosphate synthase, protein disulfide isomerase associated 3, catalase, hydroperoxide isomerase, cytosol aminopeptidase, glutamate dehydrogenase 1, 3-hydroxy-3-methylglutaryl-coenzyme A synthase 2, heat shock protein 60（Hsp60） precursor, homocysteine, 78 k Da glucose-regulated protein precursor. It was concluded that some differences were existed in the proteomic spectra expression of TCM cold and heat constitution rats. The abnormality of enzyme protein metabolism may be one of the material bases for the formation of cold and heat constitution.