| The previous experiments showed that the Echinops Latifolius Tausch Polysaccharide B(ETPB)has a significant improvement effect on blood sugar level in type 2 diabetic rats.In order to explore its mechanism,this experiment used insulin resistance rat L6 skeletal muscle cells as the model,carried out the related research on the effect and mechanism of ETPB on insulin resistance rat L6 skeletal muscle cells,and provided theoretical basis for the application of ETPB in the prevention and control of type 2 diabetes mellitus.The results show that:1.Rat L6 myoblasts were differentiated into rat L6 skeletal muscle cells after 4 days using horse serum.The optimum concentration of palmitic acid to induce insulin resistance in rat L6 skeletal muscle cells was 0.4 mmol/L in the maximum safe concentration.2.The CCK-8 method was used to determine the maximum safety concentration of ETPB for 24 hours was 200 ?g/mL,and the maximum safety concentration for 48 hours was 50 ?g/mL.ETPB action 24h could significantly increase the sugar consumption of insulin resistance rat L6 skeletal muscle cells compared with model group(P<0.05),and ETPB could significantly increase the sugar consumption of skeletal muscle cells which added to AMPK inhibitors(P<0.05).3.Using RT-qPCR to detect the expression of mRNA.Compared with the model group,ETPB could increase the expression of AMPK,GLUT4 and IRS-1(P<0.05).Using Western Blot to detect the expression of related proteins.Compared with the model group,ETPB can increase AMPK,GLUT4,IRS-1 expression(P<0.05).Conclusion:1.After palmitic acid induced rat skeletal muscle cells,sugar consumption decreased significantly,and the cells were in insulin resistance state.2.ETPB can significantly increase the sugar consumption of insulin resistance rat L6 skeletal muscle cells induced by palmitic acid.3.ETPB can improve insulin resistance of skeletal muscle cells by regulating AMPK signaling pathway-related genes and proteins. |
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