Discovery And Mechanism Of Marine Compound YK01 For Lipid Metabolism Effects

Caenorhabditis elegans is a multicellular eukaryotes.Based on the research of C.elegans,humans learned that C.elegans fat hereditary metabolic pathways,after discovery of the gene deletion,the behavior,traits and fatty acid patten of mutant C.elegans had changed and all the cell lineage had been elucidated.Therefore,it had also been included in the study of fatty acid metabolism and screening drugs important model organisms.This thesis bases on C.elegans as model of selecting marine natural products which adjust obesity,and mechanism of this compound.To adopt the method of Nile red vital staining nematodes to select marine compouds which can reduce the intensity of fluorescence.Detected the results under the fluorescence microscope,found fluorescence intensity of YKO1 group is decreased obviously.In order to verify the reliability of the Nile red experiment,we had to prove it from the aspects of biochemistry.With the method of the triglyceride and protein assay kits for determining the content of dosing group and control group in per unit protein of triglycerides.Repeat experiments found that compared dosing group to control group,triglyceride content decreased about sixteen percent.Hence YK01 indeed lower triglyceride that is the main active ingredient of fat.In order to study the mechanism of YKO1 lipid-lowering activity,and futher testing all relevant fat metabolism genes and some important transcription of the nuclear transcription factor for change.Nearly one hundred kinds of gene sequence detected that found lbp-7,lbp-8,hacd-1 genes were upregulated;fat-7,acdh-la,achd-2a were downregulated,the mechanism of YKO1 is likely effect on the several differentially expressed genes.In order to find the YKO1 lipid-lowering effect mechanism,research before and after dosing in total fatty acids composition content differences and fatty acid metabolism of key enzyme action became a key step.Using methyl ester method to make the total fatty acid methyl esterified,and GC-MS detected.The results showed that dosing group compared with control group C16:0 increased,C16:ln7 reduced,the ratio of C16:0/C16:ln7 increased 1.93 times;and the fat-5 mutant group compared with wild type group the ratio of C16:0/C16:ln7 increased 2.59 times,the two results were matched with statistical significance.According to worm fatty acid synthesis metabolic pathway,YKO1 lipid-lowering active mechanism was inhibitingfat-5 gene encoding enzyme activity of SCD.To sum up,through inhibiting SCD activity,YKO1 changed fat storage efficiency of SCD which changed the ratio of monounsaturated fatty acids and saturated fatty acid in cells,finally achieved reducing weight.