| Dietary polyphenols are a class of dietary functional ingredients that promote health and prevent a variety of chronic diseases,with a wide range of natural resource and low toxicity,which have been recognized as the most potential chemo preventive substances.Myricetin is a typical dietary flavonoid,widely distributed in a variety of fruits,vegetables,red wines and other food.The main purpose of the present thesis is to study the effects of myricetin on the expression of human genome in human hepatocytes and the inhibitory effect on macrophage inflammatory response via advanced nutrigenomics and molecular biological methods.Through the research and comparative analysis,we supply a theoretical foundation for the further application of myricetin and similar dietary polyphenol in the field of food and drug discovery,and help us to comprehensively evaluate the effect of dietary polyphenol on human health from the whole genome and proteome level.The research contents and results are shown as following.First,HepG2 cells were pre-cultured with the concentration of 1×10~6 cells/dish for 24 h,and were treated with 20?M of myricetin for additional 8 h.The total RNA was extracted and reverse transcribed into cDNA for gene chip hybridization and signal detection,the specialized microarray data analysis software(GO and IPA)were applied to analyze the large amount of gene chip data:1.There are 619 genes of the total 44k genes significantly disturbed(larger than or equal to 2 folds)by myricetin treatment in HepG2 cells,while IPA condensed them into 337 genes.The change fold range of most of the above genes is 2-3,and the number of genes larger than 2 folds the number of different expression changes is selected to count the number of genes included in each expression variation range and the source data And IPA system screening data to compare and found that differentially expressed genes are mostly up or down to change 2-3 folds,which is containing 534 genes in microarray and 286 genes in IPA analysis;2,Gene Ontology(GO)analysis reveals that these significant disturbed genes are belonging to 13,18,and 29 minor classifications of the major 3 classifications,biological processes,cellular component,and molecular function,respectively.The major classification of molecular function and the minor classification of protein binding contained the most significant disturbed genes,which indicated that myricetin significantly affect the molecular function and cellular signaling transduction is most likely to be disturbed.3.In the gene expressions of metabolizing enzymes,the most significantly affected are transporters,including 10 in SLC family,6 in ATP family,and 2 in ABC family,which implied that myricetin can significantly induce the gene expressions of hepatocyte metabolizing enzymes,especially the energy metabolism-related genes;4.IPA analysis results revealed that,the most significantly affected signalings by myricetin treatment are Metabolism of Xenobiotics by Cytochrome P450,Bile Acid Biosynthesis,and NRF2-mediated Oxidative Stress Response,and the most likely molecular function of myricetin is the improvement on Cardiovascular Disease,Metabolic Disease and Lipid Metabolism.Secondly,we had established the RAW264.7 macrophage inflammatory model by lipopolysaccharide(LPS)treatment,and performed Bio-plex cytokines assay to detect the typical 23 inflammatory cytokines simultaneously.The results found that the secretions of 9 of the total 23 cytokines in macrophages were significantly inhibited by myricetin treatment. |
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