Objective      To explore the effect of insulin on glucose uptake in H9c2 myocardial cells and its relationship with PI

3

K-AKT pathway

.       Method      A fluorescence microplate reader was used to determine the optimal concentration and incubation time of insulin for H9c2 cells. The H9c2 cells were divided to 5 groups: a normal control group

a 2-[N-(7-nitrobenz-2-oxa-1

3-diaxol-4-yl)amino

]

-2-deoxyglucose (2-NBDG) group

an insulin group

an insulin+LY294002 (PI

3

K-AKT pathway inhibitor) group

and an insulin+BIRB796 (p38MAPK pathway inhibitor) group. The glucose uptake was analyzed with the fluorescence microplate reader and a flow cytometer. The expression of glucose tansporter-1 (GLUT1) and glucose tansporter-4 (GLUT4) in the H9c2 cells was detected by indirect immunofluorescence assay and confocal laser scanning analysis.       Results      2-NBDG could be used to detect glucose uptake in H9c2 cells. The optimal concentration and incubation time of insulin were 200 μmol/L and 30 min

respectively. Insulin could promote glucose uptake in the H9c2 cells (P0.05)

and this effect was blocked by LY294002 (P0.05) but not by BIRB796 (P0.05). GLUT1 and GLUT4 were expressed in the H9c2 cell membranes

and insulin increased the expressions of GLUT1and GLUT4.       Conclusion      Insulin can increase the expression of GLUT1and GLUT4 via PI

3

K-AKT pathway rather than p38MAPK pathway

and then promote glucose uptake in the H9c2 cells.