Intervention Study Of Piperine On AMPK Signaling Pathway Downstream Targets In Insulin Resistance Model Cells

Objective:In this study,Hep G2 human hepatoma cell line and C2C12 mouse myoblast cell line insulin resistance(IR)model were established.To explore the effects of PIP in improving glucose metabolism and possible molecular pharmacological mechanisms,the expression of c AMP response element–binding protein 2(TORC2)in hepatocyte and the expression of Glucose transporter4(GLUT4)by the regulate of AMPK(AMP-activated protein kinase)and explore the differences of peroxisome proliferator activated receptor γ coactivator-1α(PGC-1α)in both cells were investigated.Methods:1.Observing the effects of PIP on the improvement glucose metabolism disorder of Hep G2 insulin resistant model cells and the regulation of related factors m RNA and protein expression.Concentration of 1μM DEX was used to establish the insulin resistance model cells of Hep G2 and classify model cells by model groups,positive control groups(ROG and Met),PIP administration groups culture cells then tested the glucose consumption in every groups.Collected cells after detection to investigate the m RNA expression of AMPKα、TORC2、PGC-1α and western blot method tested the protein expression of AMPKα、TORC2、PGC-1α.2.Observing the effects of PIP on the improvement glucose metabolism disorder of C2C12 insulin resistant model cells and the regulation of related factors m RNA and protein expression.C2C12 myoblast was cultured about 70%-80% in 6-well plates,replace complete medium into 2% horse serum culture 7-10 days.Using concentration of 0.5m M PA establish the insulin resistance model cells of C2C12 and classify model cells by model groups,positive control groups(ROG and Met),PIP administration groups culture cells then tested the glucose consumption in every groups.Collected cells after detection to investigate the m RNA expression of AMPKα、PGC-1α、GLUT4 and western blot method tested the protein expression of AMPKα、PGC-1α、GLUT4.Results:1.Concentration of 1μM DEX medium culture Hep G2 cells 48 hours established insulin resistant model cells.Positive medicine and PIP could upregulated them glucose consumption.2.Concentration of 0.5m M PA medium culture C2C12 myotube cells 16 hours established insulin resistant model cells.Positive medicine and PIP could upregulated them glucose consumption.3.These five concentrations of PIP on the HepG2 IR model for different times,it was to observe PIP improved the expression of AMPKα at the same time the expression of TORC2 and PGC-1α was decrease.4.Compared with these five concentrations of PIP on the Hep G2 IR model for different times and relate factors,it was to observe the dose and time effect relationship of PIP.10μM PIP was acted on IR cell model for 24 h,then the changes of the indexes were the most obvious.5.These five concentrations of PIP on the C2C12 IR model for different times,it was to observe PIP increased the m RNA and protein expression of AMPKα、PGC-1α、GLUT4.6.Compared with these five concentrations of PIP on the C2C12 IR model for different times and relate factors,it was to observe the dose and time effect relationship of PIP.10μM PIP was acted on IR cell model for 36 h the protein expression of GLUT4 increased significantly but 5μM PIP was acted on IR cell model for 36 h the protein expression of PGC-1α increased significantly.Conclusion:1.PIP can improve the glucose metabolism disorder in the IR model of Hep G2 cells and C2C12 cells by increased the impression of AMPKα to activated he AMPK signaling pathway.2.After activation of AMPK signaling pathway the m RNA and protein expression of TORC2、 PGC-1α was decrease in Hep G2 cells.The m RNA and protein expression of PGC-1α、GLUT4 was increase in C2C12 cells.These factors was closely related to insulin resistant and investigate it could clarify PIP improve the glucose metabolism disorde in insulin resistant.