Study On Action Mechanism And Material Base Of Compound Danshen Dripping Pills In Treatment Of Carotid Atherosclerosis Based On Techniques Of Gene Expression Profile And Modularity Analysis

In this thesis, a 2-layered heterogeneous network(2-HN) module of Compound Danshen dripping pills(DSP) was constructed using techniques of gene expression profile, modularity analysis, and techniques of molecular fingerprint. The material base and molecular action mechanism of DSP in the treatment of atherosclerosis were initially revealed by means of further analysis of the network. Significance analysis of microarray(SAM) was applied to screen out differential genes between gene expression profiles of atherosclerosis pathological tissues normal tissues. Then, the differential genes were analyzed by employing Gene Ontology(GO), KEGG pathway and GSEA analysis in DAVID; Selected differential genes were imputed to Cmap, in which drug molecules with higher negative enrichment score were obtained; At last, 336 ingredients of DSP were compared with 7 drug molecules in 2D molecular fingerprints method. A 2-layered heterogeneous network(2-HN) module was established. The 2-HN was analyzed using Girvan- Newman algorithm, in which modules were established and Modularity measures and Ratios of preserved CPIs of each modules were calculated after, and then primary pharmacological units in the evaluated key modules were investigated and analyzed employing Gene Ontology(GO) and KEGG pathway analysis; And at last, potential bioactive compounds, interacting targets and pathways in the process of treating or intervening diseases of DSP were analyzed and picked out by combining gene expression profile. A 2-layered heterogeneous network(2-HN) module was established in this thesis, and bioactive compounds and its active pathway of DSP treating or intervening atherosclerosis were predicted based on network pharmacology, which expressed the "network target, multi-components" character of TCM. Furthermore, predicted results were basically identical with existing researches and reports, which indicate that research techniques implied in this thesis were respectively reasonable and accurate, provided a methodology reference for uncovering the material base and action mechanism of TCM prescriptions.The main research contents were as follows:1. Study on the material base and molecular action mechanism of DSP in the treatment of atherosclerosis based on gene expression profile and molecular fingerprint technology: First, the data of gene expression profiles of atherosclerosis and normal tissues in human body were collected, and then SAM was applied to screen out differential genes. Then, the differential genes were analyzed by employing Gene Ontology(GO) and KEGG pathway analysis in DAVID, which is a platform of bioinformatics resources that consists of analytic tools; Some low expressed genes that possess biological significance could be picked up through GSEA analysis; Selected differential genes were imputed to Cmap, in which seven drug molecules were with higher negative enrichment score were obtained, implying that the gene expression profiles of pathological tissues could be reversely regulated by these ingredients; At last, based on the hypotheses that similar structures have similar activities, 336 ingredients of DSP were compared with 7 drug molecules in 2D molecular fingerprints method. The results showed that 147 differential genes including 60 up-regulated genes and 87 down regulated genes were screened out by SAM. And in 7 drug molecules, only isotretinoin does not possesses treatment function on cardiovascular diseases, and TTNPB have similar structure with 36 ingredients in DSP(with Tanimoto coefficients greater than 0.7), implying that main effective constituents for treating atherosclerosis in DSP are possibly in these 36 ingredients.2. Study on material base and action mechanism of DSP for treatment of atherosclerosis based on modularity analysis In this part, compound-compound interaction(CCI), compound-protein interaction(CPI) and protein-protein interaction(PPI) networks were established, and a 2-HN module of DSP was established based on that. 2-HN was analyzed by GirvanNewman algorithm applying cluster Maker plug-in in Cytoscape, and key module were evaluated by calculating the Modularity measures and Ratios of preserved CPIs of each modules, and were investigated and analyzed employing GO and KEGG pathway analysis at last. The results showed that 6 main ingredients and 5 key modules were evaluated using modularity analysis, and material base and molecular action mechanism of DSP in the treatment of atherosclerosis were initially revealed, which were were basically identical with existing researches and reports.3. Summary Combining the results of two parts, it has been found that eight ingredients in 36 ingredients which could reversely regulate atherosclerosis tissues, including Saloilenone、Danshenol A、Danshenol B、Cryptotanshinone、Tanshinone Ⅵ、Tanshinone ⅡA、Protocatechuic acid and Salvianolic acid a, were identical with module 1 and 3. Furthermore, five ingredients were found during pathway analysis in the graph, in which Cryptotanshinone, Tanshinone Ⅵ, Tanshinone ⅡA and Protocatechuic acid was in module 1, Salvianolic acid a was in module 3. Through verification by references, only Protocatechuic acid was not a bioactive compound of Salvia miltiorrhiza. Results of this thesis were basically identical with existing researches and reports, but the method is still imperfect, such as insufficient number of gene expression profile and picked-out similar structures, none disease-intervening ingredients were found in results, and respectively none obvious bioactive compounds in the intervene pathway graph. The method is worthily to be studied in sequential researches.