Palmitate-activated macrophages confer insulin resistance to muscle cells by a mechanism involving protein kinase C θ and ε

• Published in: PloS one Vol. 6; no. 10; p. e26947
• Main Authors: Kewalramani, Girish, Fink, Lisbeth Nielsen, Asadi, Farzad, Klip, Amira
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 2011; Public Library of Science (PLoS)
• Subjects: Acid resistance; Activation; Adipocytes; Animals; Biology; c-Jun protein; Cell Line; Chemokines; Conditioning; Culture Media, Conditioned - pharmacology; Cytokines; Diabetes; Fatty Acids; Glucose; Glucose - metabolism; Immunoglobulins; Inflammation; Inflammatory response; Inhibitors; Insulin; Insulin receptor substrate 1; Insulin Resistance; Isoenzymes - metabolism; Isoforms; JNK protein; Kinases; Macrophages; Macrophages - drug effects; Macrophages - metabolism; Mice; Muscle Cells - drug effects; Muscle Cells - metabolism; Musculoskeletal system; Myoblasts; Obesity; Palmitates - pharmacology; Palmitic acid; Phosphorylation; Protein kinase C; Protein Kinase C - metabolism; Protein Kinase C-epsilon - metabolism; Protein Kinase C-theta; Proteins; Ribonucleic acid; RNA; RNA-mediated interference; Rodents; Signaling; Substrate inhibition; Tissues; Transcription factors; Translocation; Tumor necrosis factor-TNF; Tyrosine; Ontario Canada; Canada
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0026947

Macrophage-derived factors contribute to whole-body insulin resistance, partly by impinging on metabolically active tissues. As proof of principle for this interaction, conditioned medium from macrophages treated with palmitate (CM-PA) reduces insulin action and glucose uptake in muscle cells. However, the mechanism whereby CM-PA confers this negative response onto muscle cells remains unknown. L6-GLUT4myc myoblasts were exposed for 24 h to palmitate-free conditioned medium from RAW 264.7 macrophages pre-treated with 0.5 mM palmitate for 6 h. This palmitate-free CM-PA, containing selective cytokines and chemokines, inhibited myoblast insulin-stimulated insulin receptor substrate 1 (IRS1) tyrosine phosphorylation, AS160 phosphorylation, GLUT4 translocation and glucose uptake. These effects were accompanied by a rise in c-Jun N-terminal kinase (JNK) activation, degradation of Inhibitor of κBα (IκBα), and elevated expression of proinflammatory cytokines in myoblasts. Notably, CM-PA caused IRS1 phosphorylation on Ser1101, and phosphorylation of novel PKCθ and ε. Co-incubation of myoblasts with CM-PA and the novel and conventional PKC inhibitor Gö6983 (but not with the conventional PKC inhibitor Gö6976) prevented PKCθ and ε activation, JNK phosphorylation, restored IκBα mass and reduced proinflammatory cytokine production. Gö6983 also restored insulin signalling and glucose uptake in myoblasts. Moreover, co-silencing both novel PKC θ and ε isoforms in myoblasts by RNA interference, but not their individual silencing, prevented the inflammatory response and restored insulin sensitivity to CM-PA-treated myoblasts. The results suggest that the block in muscle insulin action caused by CM-PA is mediated by novel PKCθ and PKCε. This study re-establishes the participation of macrophages as a relay in the action of fatty acids on muscle cells, and further identifies PKCθ and PKCε as key elements in the inflammatory and insulin resistance responses of muscle cells to macrophage products. Furthermore, it portrays these PKC isoforms as potential targets for the treatment of fatty acid-induced, inflammation-linked insulin resistance.