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Coronary heart disease is serious harm to human health of a class of diseases, \Proteins as active macromolecules in the final product of the gene and cells, its study of cardiovascular disease will prompt more complete metabolic regulation as well as the regularity of serious diseases; may be in the field of traditional Chinese medicine TCM their biological background and objective basis. Objective: To establish a serum-dimensional gel electrophoresis experimental conditions, comparison between the different stages and Coronary Heart Disease patients and normal serum protein expression differences of coronary heart disease, and find associated with coronary heart disease and Coronary Heart specific proteins, to explore the development of coronary heart disease mechanism and Coronary Heart molecular biology essence, while observing the effects of coronary heart disease Serum umbilical vein endothelial cell proliferation. : 1 coronary heart disease at different stages of the serum and blood stasis Proteomics 1.1 in strict accordance with the inclusion criteria and exclusion criteria collected four cases, select coronary heart disease in patients with various stages of each eight cases, in which the the ④ group of patients with coronary heart disease TCM syndrome are stasis syndrome, early morning collection of peripheral venous blood in each group 3ml, centrifuged to obtain serum samples in each group. 1.2 Brandford method for the determination of serum total protein concentration, and then separation of two-dimensional gel electrophoresis of serum coronary heart disease. The focus of the first to point. (IEF) electrophoresis IPGphor isoelectric focusing instrument, sample volume 1mg, select the pH range 3-10NL IPG dry strip length 24cm; then after two steps of 15 minutes each balance , the strip is transferred to a concentration of 12%, 25.5cmx20.5cmxlmm size of the SDS-polyacrylamide gel (SDS-PAGE); Coomassie blue staining, the gel after scanning the Imagemaster and PDQuest software of two groups of gel image matching, analysis, to identify differences between groups. Cut from the gel point of difference, hydrolyzed by trypsin MOLDI-TOF-MS detection point information of the resulting protein by the Mascot software searching Swissprot and NCBInr database for protein identification. Recovery of coronary heart disease each serum of HUVEC 2.1 umbilical vein endothelial cells were cultured at 37 ° C with 10% fetal bovine serum (FBS) medium? 2 concentration incubator culture. 2.2 at 3000 cells / well seeded in 96 well plates, were treated with 4 serum 0.22um membrane filter sterilization, with DMEM medium mixed in proportion of 5% serum-free DMEM culture medium, respectively, each of the groups of serum added to 96-well plates, each group of at least six wells 2.3 respectively in culture to 24h, 48h, 72h, 96h, MTT method at each hole 490nm absorbance value (OD). Results: 1 coronary heart disease at different stages of the serum and blood stasis proteomics 1.1 get ① normal healthy group, the the ② coronary heart disease in high-risk population groups, ③ The coronary heart disease in patients with acute group and ④ coronary heart disease patients with stable group average protein spots were 153.6 ± 27.57,148.8 ± 23.15,120.8 ± 6.57,146.6 ± 19.99. 1.2 by analyzing and comparing the differences between the four groups of proteins point 13, by peptide mass fingerprinting identification, query the database, the last seven meaningful protein spots were: retinol-binding protein 4 (RBP4), apolipoprotein E (ApoE), apolipoprotein A1 (ApoA1), CD5 antigen-like protein (CD 5L), haptoglobin (H p ), cluster protein (CLU), and serum albumin (ALB). 1.3 by analyzing and comparing coronary heart blood stasis group with normal human serum protein expression differences, there are six proteins differ: RBP4 of ApoE, ApoA1, CD5L,,, Hp, ALB, coronary heart disease each period serum of HUVEC whether plus or not plus 5% fetal bovine serum of four groups, four groups of serum on the proliferation of human umbilical vein endothelial cells influence the trend is the same: ①, ②, ③, ④ serum effect on the proliferation of HUVEC was gradually Enhanced trends, the strongest the ④ group promoting proliferation. Conclusion: establish a reproducible, more stable experimental conditions of the two-dimensional gel electrophoresis of serum and get better coronary heart disease serum protein expression profiling. 2 found retinol binding protein 4 (RBP4), apolipoprotein E (ApoE), apolipoprotein A1 (ApoA1), CD5 antigen-like protein (CD5L), haptoglobin protein (Hp) cluster protein (CLU) and serum albumin (ALB) and coronary heart disease-related differences in proteins, which are associated with Coronary Heart blood stasis may occur associated with the inflammatory response and lipid metabolism disorders, pending further validation . The 3 Quad serum crude HUVEC proliferation was enhanced gradually, that may contain certain cytokines promote the regeneration of vascular endothelial cells or protein components in the serum of patients with coronary heart disease, and promote the establishment of collateral circulation vascular ischemia occurs
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