Study On The Antidiabetic Activities And Mechanism Of Flavonoid Derivatives

Objective:To study the effects of flavonoid derivatives which are potential antidiabetic compounds on GLUT4 translocation and its cell signal mechanism in mouse C2C12-GLUT4myc and rat L6-GLUT4myc skeletal muscle cells.To explore the change of gene expression in mouse C2C12-GLUT4myc and rat L6-GLUT4myc skeletal muscle cells with the tiliroside-derivatives.Methods:1.L6-GLUT4myc myotube and C2C12-GLUT4myc myotube were incubated with flavonoid derivatives which have been synthesized in our laboratory,the cell surface GLUT4myc levels were detected by ELISA.2.The growth of L6-GLUT4myc rat skeletal muscle cells and C2C12-GLUT4myc mouse skeletal muscle cells with different concentrations of the active compounds was detected by MTT.3.We detected the phosphorylation of 5'-AMP-activated protein kinase and GLUT4 in mouse C2C12-GLUT4myc and rat L6-GLUT4myc skeletal muscle cells were which were incubated with active compounds.4.To investigate the mechanism of strong active compounds stimulated GLUT4myc translocation,we detected the phosphorylation of 5'-AMP-activated protein kinase,Acetyl-CoA carboxylase,protein kinase B by western blotting.5.To study the relationship between the strong activity flavone derivatives with 5'-AMP-activated protein kinase and insulin pathway,we searched the experimental of flavone derivatives with specific 5'-AMP-activated protein kinase inhibitor Compound C and insulin.6.Real time-polymerase chain reaction to detect the expression of Peroxiaome proliferator-activated receptor-a,Peroxiaome proliferator-activated receptor-y,Peroxiaome proliferator-activated receptor-?,Fatty Acid Synthese mRNA in mouse C2C12 and rat L6 skeletal muscle cells with the strong active compounds.Results:1.Flavonoid derivatives(D1-D22)can increased cell surface GLUT4myc levels in L6-GLUT4/myc myotubes differently.In rat skeletal muscle cells,Dl,D8,D18 enhanced GLUT4myc translocation in a dose and time-dependent manner.2.L6-GLUT4myc cell viability was not affected by derivative Dl,D8,D18 with 10?w/mL,and C2C12-GLUT4myc cell viability was not affected by derivative Dl,D8,D18 with 1 ?g/mL.3.The phosphorylation of 5'-AMP-activated protein kinase and GLUT4 in mouse C2C12-GLUT4myc and rat L6-GLUT4myc skeletal muscle cells were not affected by D1,D8,D18.4.The phosphorylation of 5'-AMP-activated protein kinase,acetyl-CoA carboxylase was raised by D1,D8,D18 in C2C12-GLUT4myc and L6-GLUT4myc cells(p<0.05),however,Akt phosphorylation was not affected in both cells.5.D1,D8,D18 and insulin superimposed experimental results show that GLUT4myc translocation effect having a synergistic effect in L6-GLUT4myc cells.5'-AMP-activated protein kinase specific inhibitor Compound C significantly inhibited Dl,D8,D18 GLUT4myc translocation promoting effect in both cells(p<0.05).6.D18 increase the gene expression of Peroxiaome proliferator-activated receptor-a,Peroxiaome proliferator-activated receptor-? and decrease the gene expression of Peroxiaome proliferator-activated receptor-?,Fatty Acid Synthese mRNA.in mouse C2C12 and rat L6 skeletal.muscle cells.D1,D8,CN did not change the gene expression of Peroxiaome proliferator-activated receptor-?,Peroxiaome proliferator-activated receptor-?,Peroxiaome proliferator-activated receptor-y,Fatty Acid Synthese mRNA in mouse C2C12 and rat L6 skeletal muscle cells.Conclusion:1.D1,D8,D18 increases L6-GLUT4myc and C2C12-GLUT4myc cells surface GLUT4myc levels,i.e.stimulates GLUT4myc translocation.2.D1,D8,D18 did not affect the expression of GLUT4 and 5'-AMP-activated protein kinase in skeletal muscle cells.D1,D8,D18 stimulates phosphorylation of 5'-AMP-activated protein kinase and acetyl-CoA carboxylase of C2C12-GLUT4myc and L6-GLUT4myc myotubes,Akt phosphorylation was not affected in both cells.3.D1,D8,D18 and insulin superimposed experimental results show that flavonoid derivatives and insulin increase the GLUT4myc translocation is synergistic effect.4.Compound C inhibits Dl,D8,D18 stimulated GLUT4myc translocation in skeletal muscle cells,prompts that flavonoid derivatives promote the translocation of GLUT4 were mediated by AMPK pathway.5.D18 increase the gene expression of Peroxiaome proliferator-activated receptor-a,Peroxiaome proliferator-activated receptor-? and decrease the gene expression of Peroxiaome proliferator-activated receptor-?,Fatty Acid Synthese mRNA in mouse C2C12 and rat L6 skeletal muscle cells.