| Incidence of diabetes increased year by year,changes in the pancreas cause insulin dysfunction due to a variety of unhealthy living habits.Human body excessive intake sugars compounds hardly for insulin to decomposite,excess energy stored in insulin target organs,causing chronic inflammation and insulin resistance,in the long term develop to diabetes.Bioactive ingredients derived from natural products to regulate blood sugar has multiple targets rich resources of low cost good curative effect,low toxicity and no obvious side effects,for prevention and control of diabetes has been a hot research topic.In this study,the extraction and purification process of flavone of Chaenomeles speciosa was optimized,and solid preparation with good taste and hypoglycemic and lipid-lowering function was developed.By establishing a high-fat feed combined with STZ-induced type II diabetes model,its effect on glucose and lipid metabolism in type II diabetes mice was studied,and its mechanism of action was preliminarily explored.Using ultrasonic extraction method to extract Chaenomeles speciosa flavones.PB experiments were used to screen main factors that influence,response surface experiments were used to optimize the optimum conditions.Results were determined as follows: extraction time 50 min,ultrasonic power 400 W,temperature 62 ℃,solid-liquid ratio 1:27,ethanol concentration 51%.In practice,operation of the optimal conditions for validation,flavonoid yield is 3.29%±0.28%.Static adsorption filter out the optimal resin HPD826,dynamic adsorption single factor experiment used to determine fineness ratio 1:20,the sample concentration 1.4 mg/m L,the sample amount30 m L,70% ethanol elution,according to the 1.5 BV/h velocity,collect 40-80 m L components to freeze-dry.The purity of Chaenomeles speciosa flavones was26.8%±1.2%.Silica gel further purification collection methanol: methylene chloride(7:3)components,for LC-MS/MS mass spectrometry identification analysis,four flavonoids mainly include as the following: phloretin,quercetin,dimethoxy-trihydroxy flavone and hyperoside.The flavone of Chaenomeles speciosa prepared by column chromatography(purity39.3%±2.4%)was compounded with natural plant raw materials,and the best taste formula was selected based on electronic tongue and sensory evaluation.In vitro evaluation of the compound to α-glucosidase inhibitory activity is good.Take Hep G2/IR as the model,the results showed that the flavone hypoglycemic compound of Chaenomeles speciosa increased the cell survival rate to a certain extent,and increased glucose consumption so as to improved insulin resistance.The antioxidant activity ofthe compound is good,IC50 10.9 mg/m L for superoxide radical,IC50 1.8 mg/m L for ·OH,and IC50 594.1 μg/m L for ABTS radical.The characteristics of powder and water solution of solid preparation were studied.The results showed that the solid preparation had small particle size,good stability and easy storage.In aqueous solution,the uniform particle size is small and easy to dissolve,and the viscosity is small and stable.The model of type 2 diabetes induced by high fat combined with STZ was established,and the physiological and biochemical indexes were evaluated after giving the subjects,and the influencing mechanism was explored from the protein level.Compared with the model group,oral glucose tolerance and insulin content of fasting glucose and liver glycogen content were decreased in each dose group.The activity of SOD and GSH-PX in serum was increased and the content of MDA was decreased.Solid preparation reduces the content of TC,TG,LDL-C,and increases the content of HDL-C,decreases the content of leptin,increases the content of adiponectin,and controls the blood glucose level of mice by affecting the level of lipid catabolization,lower organ index to repair inflammatory enlargement and lower levels of inflammatory cytokines TNF-α and IL-6.HE staining showed that the solid preparation improved the cellular structure disorder,uneven size and enlarged space of liver and pancreas.The protein level was determined by western blot,and it was found that the solid preparation could activate the PI3K/AKT pathway to up-regulate the expression of AMPK and GSK3,inhibit the expression of SREBP-1,and then inhibit the expression of FAS.The solid preparation of Chaenomeles speciosa flavone can improve the glucose and lipid metabolism induced by high fat combined with STZ.It is of great theoretical significance for exploring the application of natural products in directional regulation of type Ⅱ diabetes and promoting the profound processing of Chaenomeles speciosa. |
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