Effect Of Different Herb Extracts On Insulin-resistant L6Cell’s Glucose Uptake Rate And Potential Mechanisms

ObjectiveWe construct mice L6insulin resistance muscle cells to study the effects of different herb components for L6cells insulin resistance.Methods1. construction of high glucose and insulin resistance model(IR-L6)We used high glucose DMEM L6muscle cells of normal rats to induce differentiation and Coomassie blue to detect differentiation staining cells. The cells after differentiation were treated with palmitic acid to build models of insulin resistance. The remaining supernatant glucose concentration was measured by glucose oxidase at different times in order to verify whether the model of insulin resistance was successfully constructed.2. The effects of different Chinese medicine for the resistance of insulin for glucose uptake in L6cellsL6rat myoblasts were divided into nine groups:normal group L6cells (NC), insulin resistance group (IR) and7different sets of experimental medicine treatment group (IR-Drug Treated). Insulin resistance L6cells were treated with different herbs post-intervention as the experimental group, insulin resistance blank cells treated with saline, followed by [3H]-2-deoxy-D-glucose ([3H]-2DG) uptake to measure glucose intake in each group cells and assess the effects of herbal for insulin in the resistance in cells metabolize glucose.3. Hypoglycemic mechanism of Chinese medicineThe expression levels of IRS1mRNA and GLUT4mRNA levels were detected by RT-PCR. The expression of tyrosine phosphorylation of IRS1protein expression and GLUT4protein expression levels were detected by western blotting method.Results1. A high glucose insulin resistance cell model was builtGlucose oxidase results showed:palmitic acid treatment after48h, cell culture supernatants IR-L6group of glucose concentration (20.75±0.19) mmol/L, which was significantly higher than that of the control L6group (17.69±0.25mmol/L)(P<0.05).2. The effection of different Chinese medicine for the resistance of insulin for glucose uptake in L6cellsAfter6h of cell culture, glucose uptake rate in IR group were significantly decreased6.4%(P<0.05) than NC cells, which gradually increased significantly with time (P<0.05). IR group and the experimental group comparison showed that:after6h of cell culture, only berberine cells glucose uptake increased significantly in the experimental group compared with IR group (P<0.05). No significant difference was among other groups in cell glucose uptake and IR group (P>0.05). After12h of cell culture, cell glucose uptake rate in herbal extracts treated by berberine, puerarin, APS, polysaccharides and emodin groups were increased significantly than that in IR group (P<0.05), and this significance gradually increased as the culture extension of time. While ginsenosides cells and Ginkgo biloba extract cells, although a slight increase in the rate of glucose uptake, but the difference was not significant (P>0.05).3. Hypoglycemic mechanism of Chinese medicineCompared with the IR group, the IRS-1mRNA and GLUT mRNA were significantly increased in TCM group (P<0.05), IRS-1tyrosine phosphorylation and protein levels GLUT protein levels were also significantly increased synchronization.ConclusionBerberine, puerarin, APS, polysaccharides and emodin can enhance IRS-1tyrosine phosphorylation to enhance insulin resistance for glucose uptake in L6cells.