The Experimental Study Of Transcriptome And Metabolome On Pathopoiesia And Outcome Of "Cold Pathogen"

BackgroundWith the theory and technology development of the post-genome and system biology, it establishes research platform on the modern molecular biology for syndrome of TCM. Omics becomes possible and new breakthrough to the research significant problems on syndrome of TCM. But omics integration need be further to the research significant problems on syndrome of TCM. So, Transcriptome and metabolome integration on pathopoiesia and outcome of "cold pathogen" utilize existing technology and methodology platform to actively meaningfully explore on research difficulty of cold syndrome now.Objective1. Common features of metabolism and transcription on pathopoiesia of "cold pathogen"2. Repair network of metabolism and transcription on hot medicine intervention "cold pathogen"Methods1. Animal model:Experimental rats were randomly divided into three groups, Normal rats fed in conventional temperature room, Rats gave2ml of distilled water irrigation stomach from the second day of molding everyday; Model rats fed in10±2℃from4to6hours every day, Rats gave2ml of distilled water irrigation stomach from the second day of molding everyday; Dosed rats fed in10±2℃from4to6hours every day, Rats gave2ml of herbal soup irrigation stomach from the second day of molding every day. The experiment continued for3weeks.12ml venous blood of rats in experiment collected,8ml blood was conserved in-20℃for the test,4ml serum was conserved in-70℃for the test.2. Biological information method:A biological function seldom due to a single cell and molecular role, most of the biological characteristics come from complex interaction in numerous cells. Net-Work construction is contemporary bioinformatics technology to solve the problem of the main method. Net-Work construction can completely and roundly reveal how various components commonly achieve a kind of biological behavior. Data on metabolism is analyzed by PCA and PLS-DA. Data is analyzed PCA by SIMCA-P11.5, and then observe the samples gathered together, discrete and outliers, then using PLS-DA observation and discrete key cause gathered difference variables.Results1. Gene expression of cwas constructed a network. The network reflects close relationship four signal pathway, they were Cytokine-cytokine receptor interaction, Endocytosis and Focal adhesion and ECM-receptor interaction, and key genes involved Gnaq、 Plcb1、Ptk2、Pik3r3、Pdgfra、Erbb2、Vegfa、IL-6、Jun、 Mmp2、 Ptga2、Itga1、Itgb6、LOC684555and Sdc2. In addition, there were antigen processing and presentation,P53signaling pathway and Cell cycle, and key genes involved Cxcr6、 Ccr5、Ccr1、Cd8a and Klrc.2. After "cold pathogen" stimulation, model rats were interposed by radix aconiti carmichaeli of the traditional Chinese medicine. Gene expression of outcome of "cold pathogen" was constructed a network. The network reflects close relationship five signal pathway, they were Phagosome, Cell adhesion molecules, Calcium signaling pathway, Cytokine-cytokine receptor interaction and Metabolism of xenobiotics by cytochrome P450, and key genes involved Itga6、tgb3、Ptk2、Pdgfra、Pik3r3、Src、 Rac1、Gna14、Plcb1、Calm3、 Gnaq、Pla2g3、Pla2gl2a、Mylpf and Jakl.3. Data of metabolism on dosed rats and model rats were analyzed by PCA and PLS-DA to reflect radix aconiti carmichaeli intervention model rats. We pay close attention to the big contribution and difference expression in two groups in order to reduce differences between the dosed group and the model and get more real and reliable results, as effective results on radix aconiti carmichaeli intervention model rats.Conclusion 1."Cold Pathogen" make inroads on skin and damage defensive Qi in human body. The early experiment simulated to explore energy metabolism changes in organism. The research result found oxidative phosphorylation pathways active and ATP6V0D1express active; animal experiment found antigen processing and presentation and cytokine who maybe a breakthrough direction on molecular mechanism of pathopoiesia and outcome of "cold pathogen".2. The key pathways on pathopoiesia of "cold pathogen" were Cytokine-cytokine receptor interaction, Endocytosis, Focal adhesion and ECM-receptor interaction, antigen processing and presentation, P53signaling pathway and Cell cycle, and there were20key genes. But antigen processing and presentation, P53signaling pathway and Cell cycle were not connected with Cytokine-cytokine receptor interaction, Endocytosis, Focal adhesion and ECM-receptor interaction, these awaited to find after expanded sample size.3. After "cold pathogen" stimulation, model rats were interposed by radix aconiti carmichaeli of the traditional Chinese medicine. Gene expression of outcome of "cold pathogen" was constructed a network. The network reflects close relationship five signal pathway, they were Phagosome, Cell adhesion molecules, Calcium signaling pathway, Cytokine-cytokine receptor interaction and Metabolism of xenobiotics by cytochrome P450, and key genes had15. Intestinal immune network for IgA production and antigen processing and presentation were not connected with Phagosome, Cell adhesion molecules, Calcium signaling pathway, Cytokine-cytokine receptor interaction and Metabolism of xenobiotics by cytochrome P450these awaited to find after expanded sample size.4."Cold pathogen" pathopoiesia main can participate in humoral immune on Th2cell. The "cold pathogen" breaks balance between Thland Th2and trigger disease. IL-6, IL-10JL-10family, chemotactic factor, FC receptor both are very active. Pathogenic mechanism of "cold pathogen" is these description.5. After "cold pathogen" stimulation, model rats were interposed by radix aconiti carmichaeli of the traditional Chinese medicine. Herb can stimulate immune function, promote hormone, vessel activated substances and the neurotransmitter secretion, strengthen signal transmission between cells and cells, cells and ECM, guide cell recognition, promote cell proliferation, differentiation and move, stimulate second messenger signaling pathways, activate the kinase system, such as phospholipase C, phosphorylating enzymes and protein kinase. And then regulate actin dependent cellular activities and improve adhesion protein induction chemotaxis to specific function of effector cell, this intervene disease development through a series of effect, and recover the body’s health.