Adiponectin is an abundant adipocyte-derived plasma protein with antiatherosclerotic effects. Vascular signal transduction by adiponectin is poorly understood and may involve 5-AMP-activated protein kinase (AMPK), cAMP signaling, and other pathways. Hyperglycemia sharply increases the production of reactive oxygen species (ROS), which play a key role in endothelial dysfunction in diabetes. Because the recombinant globular domain of human adiponectin (gAd) reduces the generation of endothelial ROS induced by oxidized LDL, we sought to determine whether adiponectin could also suppress ROS production induced by high glucose in cultured human umbilical vein endothelial cells. Incubation in 25 mmol/l glucose for 16 h increased ROS production 3.8-fold (P < 0.05), using a luminol assay. Treatment with gAd for 16 h suppressed glucose-induced ROS in a dose-dependent manner up to 81% at 300 nmol/l (P < 0.05). The AMPK activator 5-aminoimidazole-4-carboxamide-1-␤-D-ribofuranoside (AICAR; 1 mmol/l, 16 h) only partially decreased glucose-induced ROS by 22% (P < 0.05). Cell pretreatment with AMPK inhibitors, however, failed to block the effect of gAd to suppress glucose-induced ROS, suggesting that the action of gAd was independent of AMPK. Interestingly, activation of cAMP signaling by treatment with forskolin (2 mol/l) or dibutyryl-cAMP (0.5 mmol/l) reduced glucose-induced ROS generation by 43 and 67%, respectively (both P < 0.05). Incubation with the cAMP-dependent protein kinase (PKA) inhibitor H-89 (1 mol/l) fully abrogated the effect of gAd, but not that of AICAR, on ROS induced by glucose. gAd also increased cellular cAMP content by 70% in an AMPK-independent manner. Full-length adiponectin purified from a eukaryotic expression system also suppressed ROS induced by high glucose or by treatment of endothelial cells with oxidized LDL. Thus, adiponectin suppresses excess ROS production under high-glucose conditions via a cAMP/PKA-dependent pathway, an effect that has implications for vascular protection in diabetes. Diabetes 55: -1846, 2006A diponectin is an abundant adipocyte-derived circulating plasma protein with insulin-sensitizing metabolic effects and vascular protective properties (1-4). Low adiponectin levels are associated with endothelial dysfunction (5-7) and a predisposition to vascular injury (8 -11). In cultured endothelial cells, adiponectin has been shown to exhibit various anti-inflammatory effects, in particular those that counter the adverse cellular influences of increased oxidative stress or stimulation with cytokines such as tumor necrosis factor-␣ (TNF-␣) (2). Adiponectin binds to the walls of catheter-injured vessels (12,13) and inhibits the expression of several adhesion molecules, including vascular cell adhesion molecule-1, E-selectin, and intracellular adhesion molecule-1 induced by the cytokine TNF-␣, and it reverses the adhesion of human monocytic THP-1 cells to cultured endothelial cells (14) and enhances nitric oxide (NO) production by endothelial cells (15,16). Although the metabolic effec...

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