Proteomic Study On Peripheral Blood Mononuclear Cells Of Cerebral Infarction Patients With Ganyang-Huafeng Syndrome(GYHF)

Objective:1. TO establish cerebral infarction Ganyang-Huafeng Syndrome (GYHF) and Yinxu-Fengdong Syndrome(YXFD) high-resolution map of the two-dimensional gel electrophoresis;2. Using Comparison of proteomics methods to analyze and identify different proteins in cerebral infarction Ganyang-Huafeng Syndrome(GYHF) and Yinxu- Fengdong Syndrome(YXFD);3. From the gene expression of adenylate cyclase (AC) in cerebral infarction Ganyang-Huafeng Syndrome(GYHF) and Yinxu-Fengdong Syndrome(YXFD), to explore the relevance and the difference of Chinese medicine Ganyang-Huafeng Syndrome(GYHF) and Yinxu-Fengdong Syndrome(YXFD).Methods: Design 3 groups, cerebral infarction Ganyang-Huafeng Syndrome group(GYHF), Yinxu-Fengdong Syndrome group (YXFD),and healthy control group,in strict accordance with the disease diagnosis, diagnostic criteria syndrome, cerebral infarction choice of Ganyang-Huafeng Syndrome(GYHF)patients of 15 cases, Yinxu-Fengdong Syndrome (YXFD) patients of 15 cases, in accordance with the exclusion criteria included, choose healthy people in 15 cases, isolated from peripheral blood mononuclear cells extracted proteins. The two-dimensional gel electrophoresis, Coomassie brilliant blue staining patterns of access to gel, PDQuest V7.3.1 software comparative analysis of the three groups and identify differentially expressed proteins and the same expression of proteins of Ganyang-Huafeng Syndrome(GYHF)and Yinxu-Fengdong Syndrome(YXFD). MALDI-TOF-MS mass spectrometer for peptide mass fingerprint identification, Mascot retrieval software in the Swiss-prot database search for protein identification, retrieval related protein's name, classification and functions. By RT-PCR technology to detect gene expression of adenylate cyclase(AC) in the cerebral infarction Ganyang-Huafeng Syndrome(GYHF)and Yinxu-Fengdong Syndrome(YXFD).Results:1. Initially established PBMC 2-DE maps of Ganyang-Huafeng Syndrome(GYHF) patients and Yinxu-Fengdong Syndrome(YXFD) patients and healthy group, respectively proteins are 459, 552,644;2. PDQuest V7.3.1 software comparative analysis ,and found 8 same proteins and 23 different proteins in Ganyang-Huafeng Syndrome (GYHF) group and Yinxu-Fengdong Syndrome(YXFD)group.3. MALDI-TOF-MS mass Spectrometry analysis and MASCOT software retrieval Swissprot protein database identified 27 protein spots, a total of 20 kinds of proteins, various protein spots express different in different groups,compared with Ganyang-Huafeng Syndrome(GYHF) group and healthy group, Yinxu-Fengdong Syndrome(YXFD)group and healthy group, Ganyang-Huafeng Syndrome(GYHF) group and Yinxu-Fengdong Syndrome(YXFD)group,we found 23 different protein spots, 22 identified with 15 kinds of proteins, they are Capping protein, Thrombospondin-1, Adenylyl cyclase-associated protein 1, Rho GDP-dissociation inhibitor 2,γ-actin, Transgelin-2, Prohibitin, myosin regulatory light chain,Glutathione transferase omega-1, Thioredoxin-dependent peroxide reductase, Visual cortex cDNA, Hypothetical protein NEDD5, Ras suppressor protein,RAP1B protein, ZYX protein [Fragment]. 8 same protein spots, 6 identified with 5 kinds of proteins,they are Heterogeneous nuclear ribonucleo proteinK, serine/threonine-specific protein kinase, Talin-1, fibrinogen beta chain precursor, fibrinogen alpha chain precursor. Including the cytoskeleton protein, signal transduction protein, redox protein, apoptosis-related protein, blood protein, and other types of abnormal proteins.4. Expression of ACmRNA in Ganyang-Huafeng Syndrome(GYHF) group was missing, Yinxu-Fengdong Syndrome(YXFD)group was significantly down-regulated, and the expression have significant difference (P <0.05).Conclusion:1.Initially established cerebral infarction Ganyang-Huafeng Syndrome(GYHF) , Yinxu-Fengdong Syndrome(YXFD) and healthy PBMC 2-DE maps, Through MODI-TOP-MS analysis of Ganyang-Huafeng Syndrome(GYHF) , Yinxu-Fengdong Syndrome (YXFD) , the same protein expression and protein expression of differences, suggesting that the disease similar cards have a common material basis of different ,and have different connotations of the essence.2. RT-PCR verified ACmRNA expression in Ganyang-Huafeng Syndrome(GYHF) and Yinxu-Fengdong Syndrome(YXFD) ,the results is the same with protein test results, Proved the stability of Proteomics technology and the accuracy of the results.