Study On The Synergistic Hypoglycemic Mechanism Of D-chiro-inositol In Tartary Buckwheat And Polyphenols

With the increasing incidence of diabetes,it has become the third major disease affecting human health after cardiovascular and cerebrovascular diseases and malignant tumors.Among them,type 2 diabetes is up to 90%.Continuous hyperglycemia stimulation is one of the important factors in the pathogenesis of type 2 diabetes.Therefore,controlling the rise in blood sugar is the key to relieving type 2 diabetes.In this study,hypoglycemic factors were obtained from medicine food homology(MFH)and common food,the active components were further tracked,isolated and identified,and their synergistic hypoglycemic effect and hypoglycemic activity in vitro and in vivo were studied to reveal the glucose metabolism mechanism of liver,which has practical significance for adjuvant treatment of type II diabetes(T2DM).Firstly,hypoglycemic factors were screened by in vitro experiments.Tartary buckwheat DCI,green tea EGCG and hawthorn extract lower glucose levels by promoting cell glucose consumption or inhibiting ?-glucosidase activity.Secondly,the DCI extraction scheme of Tartary buckwheat was optimized by single factor,slope climbing and response surface experiments,column chromatography graded purification and high performance liquid phase analysis were used to track the active components of hawthorn and identify with LC-MSMS.Thirdly,the Chou-Talalay model was established to explore the synergistic hypoglycemic effect of DCI and polyphenols in vitro.Finally,animal model of diabetes was induced by STZ,the glucose metabolism was investigated in vivo,and the regulatory mechanism of liver PI3K/AKT/FOXO1 and GSK3 signaling pathways was studied by Western blot.In vitro experiments confirmed that Tartary buckwheat DCI,green tea EGCG and hawthorn active components were good components of increasing cell glucose consumption and ?-glucosidase inhibitor.The best DCI extraction scheme of Tartary buckwheat was as follows: 50% ethanol,p H 3,solid-liquid ratio 1:25,50?,400 W ultrasonic for 30 min.Under these conditions,the DCI yield was 2.83%.In addition,the DCI of Tartary buckwheat was purified by adsorption and desorption experiments.The purity of the Tartary buckwheat was up to 63.98% by high performance liquid phase analysis after it was successively processed by macroporous resin H103 and DEAEcellulose column.After D101 macroporous resin,silica gel and Sephadex LH-20 gel,hawthorn extract determined that the active ingredient inhibiting ?-glucosidase was HP.DCI of Tartary buckwheat and polyphenols(green tea EGCG and hawthorn HP)synergism inhibited ?-glucosidase activity(CI=0.89),promoted glucose consumption in cells,inhibited gluconeogenesis,and promoted glycogen synthesis.In vivo experiments were conducted to explore the effect of DEH(DCI,EGCG and HP synergistic complex)on glucose metabolism in STZ-induced diabetic mice.It was found that oral glucose tolerance,insulin level and insulin resistance of the mice were reduced.Serum TC,TG,FFA and LDL-C were reduced,but HDL-C content was increased,which improved abnormal lipid metabolism in diabetic mice.Moreover,serum levels of inflammatory cytokines IL-6 and TNF-? were reduced,which may reduce the incidence of inflammatory response.The levels of glycosylated hemoglobin and Leptin were decreased,and the levels of glucagon-like peptide-1 and adiponectin were increased.The activity of ALT and AST decreased,which showed the protective effect on liver cells.It inhibited the activity of small intestinal ?-glucosidase protein solution in diabetic mice,and reduced the postpranpraneal blood glucose rise caused by the hydrolysis of carbohydrates.Pathological tissue sections of liver,kidney and pancreas of mice showed that DEH restored tissue damage in diabetic mice,and PAS staining increased glycogen content and increased glucose storage.Investigate the effect of DEH on glucose metabolism in the liver of T2 MD mice.It was found that DEH activated PI3K/AKT signaling pathway,promoted FOXO1 phosphorylation inactivation,reduced PEPCK and G6 Pase activity,reduced non-glucose substance transformation,and promoted GSK3 phosphorylation inactivation,inhibited GS phosphorylation inactivation,and increased liver glucose storage.In conclusion,DCI of Tartary buckwheat and polyphenols showed synergistic hypoglycemic effect.This study provides a theoretical basis for the research and development of new compound hypoglycemic health food and drugs.