Effects Of Chrysanthemum Geranilin On Type ? Diabetic Mice And Its Proteomic Study

To screen constituents with hypoglycemic activity from plant matrices and reveal their molecular mechanism are helpful to the development of functional foods.Chrysanthemum is a well-known edible medicinal plant in China.Modern studies have shown that chrysanthemum extract and selenium polysaccharide have hypoglycemic activity,but there are few studies on the hypoglycemic effect and its mechanism of phenolic compounds in chrysanthemum.To Screen monomers with hypoglycemic activity from chrysanthemums and study their mechanism of lowering blood glucose has a positive effect on the in-depth development of chrysanthemum resources and development of health food for assisting in lowering blood glucose.In this experiment,computer simulation and two in vitro enzymatic experiments of ?-amylase and?-glucosidase were used to screen the most active monomer diosmetin from four phenolic compound monomers including isochlorogenic acid C,luteolin,linarin,and diosmetin,and then by inducing type ? diabetes mice model to study diosmetin in vivo hypoglycemic activity from muliple aspects such as fasting blood glucose(FBG),glycosylated serum protein(GSP)and glucose tolerance(OGTT),finally,the effect of high-dose diosmetin intervention on protein expression in the liver of type ? diabetic mice from the perspective of molecular biology was further studied to explore the possible molecular mechanism of diosmetin in alleviating hyperglycemia.The brief method and results of the experiment analysis are as follows:Through computer simulation and two in vitro enzymatic experiments of?-amylase and ?-glucosidase,the in vitro activities of isochlorogenic acid C,luteolin,linarin and diosmetin in reducing blood glucose were studied,and comprehensively screen a monomer with higher blood glucose lowering activity.Firstly,using?-glucosidase protein as the receptor,isochlorogenic acid C,luteoside,linarin,and diosmetin as the ligands,through the molecular docking(computer simulation)to study the docking strength of receptor protein and ligand moleculer.The experimental results showed that four phenolic compound monomers could bind well with receptor protein,the combination ability is ranked from large to small for:isochlorogenic acid C>diosmetin>luteolin>linarin.The results of in vitro enzymatic experiments showed that the IC25 values of isochlorogenic acid C,diosmetin,luteolin,and linarin for ?-amylase inhibition are:112.2?g/mL,0.6?g/mL,0.4?g/mL,1.8?g/mL,and the IC25 values of isochlorogenic acid C,diosmetin,luteolin,and linarin for ?-glucosidase inhibition are:9.6?g/mL,5.7?g/mL,8.3?g/mL,50.8?g/mL.From the above experimental results,it can be concluded that the protein binding ability of diosmetin and the inhibitory activity of diosmetin on blood glucose regulating enzymes are relatively strong.In order to study the relieving effect of diosmetin on type ? diabetes from multiple aspects,in this experiment,we first used Kunming mice to establish type ?diabetes model,and then intervened by gavaging with diosmetin,and studied its effect on the general symptoms of diabetes such as blood lipids,antioxidant indexes and organ protection in type ? diabetic mice.Experimental results show that diosmetin could improve the adverse symptoms of high food intake,high water intake and low body weight in type ? diabetic mice,and alleviates abnormal symptoms of glucose metabolism disorders such as high fasting blood glucose,insulin resistance,and abnormal oral glucose tolerance.Through the determination results of blood lipid indexes such as total cholesterol and total triglycerides,the results found that diosmetin has a good regulatory effect on lipid metabolism in type ? diabetic mice.Through the determination of antioxidant indicators such as superoxide dismutase and glutathione peroxidase,it is found that diosmetin could improve the antioxidant capacity of type ?diabetic mice.Through the measurement results of liver glycogen,it is found that diosmetin could regulate liver glucose metabolism of type ? diabetic mice and promote it glycogen synthesis.Through the experimental results of organ index,alanine aminotransferase activity,combined with liver and kidney tissue HE staining results,it could be concluded that diosmetin could alleviate liver and kidney damage caused by type ? diabetic.From the above results,it could be concluded that diosmetin played a role in relieving type ? diabetes by improving the symptoms such as the disorders of glucose and lipid metabolism,decreased antioxidant levels,liver and kidney damage in mice.In order to study the changes in protein expression in the liver of type ? mice after the intervention of high dose diosmetin(D50),this experiment used non-labeled quantitative technology to explore the possible molecular mechanism of diosmetin in the process of relieving type ? diabetes from the perspective of molecular biology.The experimental results showed that there were 133,259,and 315 differential proteins(DP)in these three comparison groups of high-dose diosmetin intervention group(D50)vs model group(M),normal group(N)vs model group(M)and high-dose diosmetin intervention group(D50)vs normal group(N).Hierarchical cluster analysis and other results showed that in the comparison of D50 vs M,glucokinase(GCK),tyrosine 3-plus monooxygenase/tryptophan 5-plus monooxygenase activating protein(YWHAB),cytochrome P450 Family member 2C70(CYP2C70)have been significantly upregulated,glycogen synthesis kinase(GSK-3?),Samds family member protein 3(Samd3),tyrosine protein kinase 1(CSNK1d),pyruvate kinase(PKM),cell Pigment P450 family member 2E1(CYP2E1)have been significantly down-regulated.Through GO annotation analysis,it is found that these proteins have a variety of biological functions and activities,and involved in multiple regulation,catalysis,and metabolic processes.The results of KEGG enrichment analysis showed that in the comparison of D50 vs M,glycolysis/gluconeogenesis pathway,Hippo signaling pathway,linoleic acid metabolism,arachidonic acid metabolism may be related to the regulation of glucose metabolism,lipid metabolism,and oxidative stress in type ? diabetes,these pathways changed significantly,and the trend of changes in the expression of DP enriched in each pathway is consistent with the previous studies.It can be inferred that diosmetin may regulate these metabolic pathways and the enriched DP(GCK and PKM in the glycolysis/gluconeogenesis pathway,GSK-3?,Samd3,YWHAB,CSNKld in the hippo signaling pathway,CYP2C70,CYP2E1 in the linoleic acid metabolism and arachidonic acid metabolism pathway)to exert the molecular mechanism of relieving type ?diabetes.