Figure 7

Vaspin induces the activation of AMPK independently of Akt and NO pathway. HAECS were incubated with 100 ng/ml vaspin or vehicle for 1 hr at 37°C. Wortmannin (100 nM, 681675, Calbiochem, Darmstadt, Germany), a specific covalent inhibitor of PI3-kinase or nitro-L-arginine (500 μM, L-NNA, N5501, Sigma), an eNOS inhibitor, was pretreated 15 min before the vaspin treatment. Phosphorylation of AMPK (A and C), and ACC (B and D) were detected by Western blotting using phospho-specific antibodies for AMPK and ACC, normalized to total AMPK and ACC. Data are shown as means ± SEM of three independent experiments. AMPK, AMP-activated protein kinase; NO, nitric oxide; ACC, acetyl-CoA carboxylase; N-NNA, nitro-L-arginine; eNOS, endothelial nitrix oxide synthase. In A and B, ^*p < 0.05 vs. untreated (Control), ^†p < 0.05 vs. cells treated with wortmannin alone. In C and D, ^*p < 0.05 vs. untreated (Control).