| Objective:In this study,we will extract the hydrolysable tannins from Rosa rugosa buds?RRB?and confirm its structure;through the establishment of animal model and cell experiment,we will make clear the their hypoglycemic activity and material basis of pharmacodynamics.Methods:1.Extraction,separation and structure determination of effective component:Add 1.5L 50%ethanol to 1500g Rosa rugosa bud powder,mix well,stand for 24h,extract three times,combine filtrate,and concentrate to about 200ml.Then extract three times with equal amount of ethyl acetate,combine,concentrate and freeze dry.The ethyl acetate extract was named by polyphenol extract of Rosa rugosa buds?PERRB?and prepared for pharmacological activity evaluation and active component separation.The PERRB?10g?was separated and purified repeatedly by column chromatography?Daiaion HP-20,Sephadex LH-20,etc.?and preparative high performance liquid chromatography?Prep-HPLC?,characterized by comprehensive analysis of experimental data such as nuclear magnetic resonance?NMR?,mass spectrometry?TOF-Mass?,etc.,to determine the structure of the isolated compounds.2.The effect of PERRB on T2DM rats:90 male SD rats?200±20g?were randomly selected as the blank group and the model group.The rats in the model group were injected with 30mg/kg STZ and the rats in the blank group were injected with the same amount of normal saline.After the model was established successfully,it was divided into model group,control group?0.0378mg/g Acarbose?,PERRB low-dose group?0.0348mg/g PERRB?,PERRB medium-dose group?0.0697mg/g PERRB?,PERRB high-dose group?0.1394mg/g PERRB?,RRB Group?0.81mg/g Rose?.After 30 days of administration,rats were fasted for 12 hours,anesthetized with 10%chloral hydrate,blood was collected from abdominal aorta,centrifuged,and serum Glu,INS,GC,GSP,TG,TC,HDL-C,LDL-C,FFA,SOD,MDA,TNF-?,IL-6,NF-KB were measured.The liver,kidney and pancreas were removed to prepare pathological sections,and gastrocnemius muscle was taken to measure GLUT4 content by Western blot.3.Effect of tellimagrandin I on IR-Hep G2cells:Through the establishment of IR model experiment,the effects of PERRB and tellimagrandin I on glucose metabolism of IR Hep G2 cells were observed.Hep G2 cells were cultured in RPMI1640 complete medium at 37?in 5%CO2 incubator,and the medium was changed every two days.When the cell adheres to the wall and grows to 90%,it is subcultured.Cell activity?number of living cells?was measured by CCK-8 kit.IR cell model was established with 1Ś10-6mol/L insulin solution.The experiment was divided into control group and administration group?Acarbose group,PERRBgroup,Tellimagrandin Igroup?,the treated cells were cultured for 24-72h.The number of cells in the supernatant was measured by CCK-8 method.The glucose content in cell supernatant was measured by Elisa method,and the glucose consumption was calculated,and the cell number per pore was calibrated.Results:1.The extraction rate of ethyl acetate extract of rose was 8.61%,129.15g.And100g were obtained for pharmacological experiment.A compond was isolated from the ethyl acetate extract of RRB by column chromatography and semi preparation.The NMR and TOF-MS data compared with the references,it were identified as tellimagrandin I.2.After 30 days of drug intervention,the general state of rats in rose and perrb groups improved,and the weight of rats gradually stabilized after 2 weeks of drug intervention.RRB and PERRB significantly reduced the serum Glu,INS,GC,GSP,TC,TG,LDL-C,MDA contents,there is a tendency to reduce the content of TNF-?and NF-?B,increased the serum HDL-C,SOD contents,improved liver fatty lesions,and effectively improved the GLUT4 expression of skeletal muscle.3.The glucose consumption of PERRB group and tellimagrandin I group increased significantly after 24 hours,48 hours and 72 hours of intervention.Conclusion:Mongolian medicine Rosa rugosa bud polyphenol extract are effective in the treatment of type 2 diabetes and tellimagrandin I was maybe its active compond. |
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