PPARβ activation restores the high glucose‐induced impairment of insulin signalling in endothelial cells

• Published in: British journal of pharmacology 2014-06, Vol.171 (12), p.3089-3102
• Main Authors: Quintela, A M, Jiménez, R, Piqueras, L, Gómez‐Guzmán, M, Haro, J, Zarzuelo, M J, Cogolludo, A, Sanz, M J, Toral, M, Romero, M, Pérez‐Vizcaíno, F, Duarte, J
• Format: Article
• Language: English
• Subjects: Animals; Blood Glucose - drug effects; Blood Glucose - metabolism; Cells, Cultured; Diabetes Mellitus, Experimental - chemically induced; Diabetes Mellitus, Experimental - drug therapy; Diabetes Mellitus, Experimental - genetics; Diabetes Mellitus, Experimental - metabolism; Endothelial Cells - drug effects; Endothelial Cells - metabolism; Enzyme Activation; Extracellular Signal-Regulated MAP Kinases - metabolism; Human Umbilical Vein Endothelial Cells - drug effects; Human Umbilical Vein Endothelial Cells - metabolism; HUVECs; Hypoglycemic Agents - pharmacology; Insulin - metabolism; insulin signalling; Male; Nitric Oxide - metabolism; Nitric Oxide Synthase Type III - metabolism; Phosphorylation; PPAR-beta - agonists; PPAR-beta - genetics; PPAR-beta - metabolism; PPARβ; Protein-Serine-Threonine Kinases - metabolism; Proto-Oncogene Proteins c-akt - metabolism; Rats, Wistar; reactive oxygen species; Reactive Oxygen Species - metabolism; Research Papers; Signal Transduction - drug effects; Time Factors; Transfection
• ISSN: 0007-1188; 1476-5381
• DOI: 10.1111/bph.12646

Background and Purpose PPARβ enhances insulin sensitivity in adipocytes and skeletal muscle cells, but its effects on insulin signalling in endothelial cells are not known. We analysed the effects of the PPARβ/δ (PPARβ) agonists, GW0742 and L165041, on impaired insulin signalling induced by high glucose in HUVECs and aortic and mesenteric arteries from diabetic rats. Experimental Approach Insulin‐stimulated NO production, Akt‐Ser473 and eNOS‐Ser1177 phosphorylation, and reactive oxygen species (ROS) production were studied in HUVECs incubated in low‐ or high‐glucose medium. Insulin‐stimulated relaxations and protein phosphorylation in vessels from streptozotocin (STZ)‐induced diabetic rats were also analysed. Key Results HUVECs incubated in high‐glucose medium showed a significant reduction in insulin‐stimulated production of NO. High glucose also reduced insulin‐induced Akt‐Ser473 and eNOS‐Ser1177 phosphorylation, increased IRS‐1‐Ser636 and ERK1/2‐Thr183‐Tyr185 phosphorylation and increased ROS production. The co‐incubation with the PPARβ agonists GW0742 or L165041 prevented all these effects induced by high glucose. In turn, the effects induced by the agonists were suppressed when HUVEC were also incubated with the PPARβ antagonist GSK0660, the pyruvate dehydrogenase kinase (PDK)4 inhibitor dichloroacetate or after knockdown of both PPARβ and PDK4 with siRNA. The ERK1/2 inhibitor PD98059, ROS scavenger catalase, inhibitor of complex II thenoyltrifluoroacetone or uncoupler of oxidative phosphorylation, carbonyl cyanide m‐chlorophenylhydrazone, also prevented glucose‐induced insulin resistance. In STZ diabetic rats, oral GW0742 also improved insulin signalling and the impaired NO‐mediated vascular relaxation. Conclusion and Implications PPARβ activation in vitro and in vivo restores the endothelial function, preserving the insulin‐Akt‐eNOS pathway impaired by high glucose, at least in part, through PDK4 activation.