| Subject:With the technology of two-dimensional polyacrylamide gel electrophoresis(2-DE),we extracted and separated total protein of Peripheral blood of the CHB patients with Internal Stasis of Dampness and Heat Syndromes(Shire Zhongzu Syndromes),Depressed Liver With the Insufficient Spleen Syndromes(Ganyu Pixu Syndromes), Collateral Obstruction by Blood Stasis Syndromes(Yuxue Zuluo Syndromes) and normal persons.And we established the expression maps of proteomics in these groups.According to analysis and comparing of the differences amone the 4 groups'proteomics express,we studied the differential proteins,which can be helpful to provide scientific evidence for approaching the nature of TCM Syndromes.Methods:To collect clinical plasma samples of patients with CHB, divided into three groups:the Middle Energizer Dampness-heat Syndromes(group S), the Liver Depression and Spleen Deficiency Syndrome(group P), and the Collateral Obstruction by Blood Syndromes(group Y), according to TCM syndrome classification standards. Control group were normal peoples(HBsAg are positive). In this experiment we established 2-DE technique successfully by optimizing a series of factors, efforts focusing on sample preparation, pH range selection, loading amount, electrophoresis parameters optimization and protein staining and so on. High resolution reproduction 2-DE maps were obtained when the pH4-7 IPG gels were loaded with about 130μg proteins each, and run under 75000v.h. After scanning the stained 2-DE gels, we use the ImageMaster software to analysis the differential protein spots between the TCM groups and normal persons. MALDI-TOF-MS and searching in SWISS-PORT database were used to identify proteins.Results:The expression maps of proteomics including good resolution and reproducibility of the CHB patients with Group S,Group P,Group Y and the normal persons were obtained after sliver staining.The average spots'number of Peripheral blood total protein is (290±17) in healthy persons, (350±23) in CHB Disease patients with Shire Zhongzu Syndromes, (328±14) in CHB Disease patients with Ganyu Pixu Syndromes and (489±25) in CHB Disease patients with Yuxue Zuluo Syndromes.(2)We selected 13 spots with differential expression to make mass-spectrum identification, and identified 10 proteins.The function of the initially identified proteins related to immune disorder, abnormal lipid metabolism and inflammatory response, etc. Compared with the normal persons,there are 5 up-regulated proteins and 5 down-regulated proteins in Internal Stasis of Dampness and Heat Syndromes. There are 4 up-regulated proteins and 6 down-regulated proteins in Depressed Liver With the Insufficient Spleen Syndromes. There are 4 up-regulated proteins and 6 down-regulated proteins in Collateral Obstruction by Blood Stasis Syndromes. Conclusion:(1)The expression maps of proteomics of CHB patients with Shire Zhongzu Syndromes and Ganyu Pixu Syndromes, Yuxue Zuluo Syndromes, and the healthy persons were established.(2)The polyprotein differencial expression between syndromes suggests that have different material foundation (different proteins) in a same disease with different TCM syndromes. This result settle scientific ground for essential research of TCM syndromes.(3) The differencial expression protein in Internal Stasis of Dampness and Heat Syndromes related to the abnormal lipid metabolism. The apoC increased expression maybe was the result of cell membrane damage significantly and binding sites(receptor) changes. (4)The proteins represented liver repair and secretion function decreased expression in Depressed Liver With the Insufficient Spleen Syndromes compared to the normal persons, but slightly higher than other groups.(5)The IGJ,SAMP and HPT are closely related to syndrome evolution.
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