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Study on the Anti-diabetes Activities of Tiliroside Derivantives and Its Mechanism

Author: ZhouZuo
Tutor: DuanHongQuan
School: Tianjin Medical University
Course: Pharmacology
Keywords: Tiliroside Derivative IR-HepG2 IR-3T3-L1 glucose consumption glucose uptake AMPK ACC
CLC: R285.5
Type: PhD thesis
Year: 2013
Downloads: 153
Quote: 0
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Objective:To establish the insulin resistant HepG2and3T3-L1adipocyte cells model. Screening of tiliroside derivantives with anti-diabetes activities. Objective to investigate the mechanism.Methods:1. To set up insulin resistant cell model:Insulin resistant HepG2and3T3-L1adipocyte cells model are commonly used in the research on insulin resistance. Copy the insulin resistant cell model, whose formation was assessed by detecting the ability of uptaking glucose with GOD-POD assay2. Evaluation of the anti-diabetes activity of the derivatives:Tiliroside was the main anti-diabetic constituent isolated from Potentilla Chinese Ser. For the improvement of its bioactivity, we have synthesized a series of tiliroside-derivatives. All the derivatives were investgated for the glucose consumption of insulin resistance HepG2and3T3-L1adipocyte cells model to evaluate the anti-diabetes activities.3. Study on mechanism of anti-diabetes activity:To investigate the mechanism of Comp.5f, we detected the phosphorylation of5’-AMP-activated protein kinase (AMPK), Acetyl-CoA carboxylase (ACC), protein kinase B (Akt) by western blotting. We also detect the effect of glucose transporters2and glucose transporters4by western blotting.Results:1. The insulin resistant HepG2cell model was established successfully by high concentration of insulin (5×10-7mol/l) in vitro culture. The IR-3T3-L1adipocyte cell model was established successfully by dexamethasone (1μmol/1,4days). Detecting the ability of uptaking glucose with GOD-POD assay. There were significant differences (p<0.05) compared with the control group. The insulin resistant cell models were established successfully.2. All of the derivatives could improve the glucose consumption of the insulin resistance cell except7b. Compounds5b、5e、5g and10b revealed significant activities compared with that of market drug, Metformin. Compounds5f、7d、10c showed stronger activities than Metformin.Among tiliroside derivantives compounds5a、5g、5f、5e、7d, Compounds5f is the most active derivatives. Compounds5f strongly2-deoxidation-[3H]-glucose uptaking in3T3-L1adipocyte cell.Compounds5f strongly phosphorylated AMPK and its substrate, ACC in both HepG2and3T3-L1adipocyte cells (p<0.05). However, Akt phosphorylation was not affected in both cells. GLUT2can be down regulated by Compounds5f in HepG2cells. Compounds5f did not affect the expression of GLUT4and AMPKa in3T3-L1adipocyte cells.Conclusion:The insulin resistant HepG2and3T3-L1adipocyte cell models were established successfully. Tiliroside derivatives have relatively stronger anti-diabetes activities. Compounds5f strongly phosphorylated AMPK and its substrate, ACC in both HepG2and3T3-L1adipocyte cells (p<0.05). Its mechanism involving AMPK correlation signal pathways.

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