title: Restoring AS160 phosphorylation rescues skeletal muscle insulin resistance and fatty acid oxidation while not reducing intramuscular lipids.
abstract: We examined whether AICAR or leptin rapidly rescued skeletal muscle insulin resistance via increased palmitate oxidation, reductions in intramuscular lipids, and/or restoration of insulin-stimulated AS60 phosphorylation. Incubation with palmitate (2 mM, 0-18 h) induced insulin resistance in soleus muscle. From 12-18 h, palmitate was removed or AICAR or leptin was provided while 2 mM palmitate was maintained. Palmitate oxidation, intramuscular triacylglycerol, diacylglycerol, ceramide, AMPK phosphorylation, basal and insulin-stimulated glucose transport, plasmalemmal GLUT4, and Akt and AS160 phosphorylation were examined at 0, 6, 12, and 18 h. Palmitate treatment (12 h) increased intramuscular lipids (triacylglycerol +54%, diacylglycerol +11%, total ceramide +18%, C16:0 ceramide +60%) and AMPK phosphorylation (+118%), whereas it reduced fatty acid oxidation (-60%) and insulin-stimulated glucose transport (-70%), GLUT4 translocation (-50%), and AS160 phosphorylation (-40%). Palmitate removal did not rescue insulin resistance or associated parameters. The AICAR and leptin treatments did not consistently reduce intramuscular lipids, but they did rescue palmitate oxidation and insulin-stimulated glucose transport, GLUT4 translocation, and AS160 phosphorylation. Increased AMPK phosphorylation was associated with these improvements only when AICAR and leptin were present. Hence, across all experiments, AMPK phosphorylation did not correlate with any parameters. In contrast, palmitate oxidation and insulin-stimulated AS160 phosphorylation were highly correlated (r = 0.83). We speculate that AICAR and leptin activate both of these processes concomitantly, involving activation of unknown kinases in addition to AMPK. In conclusion, despite the maintenance of high concentrations of palmitate (2 mM), as well as increased concentrations of intramuscular lipids (triacylglycerol, diacylglycerol, and ceramide), the rapid AICAR- and leptin-mediated rescue of palmitate-induced insulin resistance is attributable to the restoration of insulin-stimulated AS160 phosphorylation and GLUT4 translocation.
authors.affiliation: Dept. of Human Health and Nutritional Sciences, Univ. of Guelph, Guelph, ON, N1G 2W1, Canada.
authors.forename: Hakam
authors.initials: H
authors.lastname: Alkhateeb
authors.forename: Adrian
authors.initials: A
authors.lastname: Chabowski
authors.forename: Jan F C
authors.initials: JF
authors.lastname: Glatz
authors.forename: Brendon
authors.initials: B
authors.lastname: Gurd
authors.forename: Joost J F P
authors.initials: JJ
authors.lastname: Luiken
authors.forename: Arend
authors.initials: A
authors.lastname: Bonen
mesh_terms: D000263:Adenylate Kinase / Q000378:metabolism; D000620:Aminoimidazole Carboxamide / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D015153:Blotting, Western; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002518:Ceramides / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism*; D020690:GTPase-Activating Proteins / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D051275:Glucose Transporter Type 4 / Q000378:metabolism; D005989:Glycerides / Q000378:metabolism; D066298:In Vitro Techniques; D007328:Insulin / Q000494:pharmacology; D007333:Insulin Resistance / Q000502:physiology*; D020738:Leptin / Q000494:pharmacology; D050356:Lipid Metabolism / Q000502:physiology*; D008297:Male; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism*; D051058:Oncogene Protein v-akt / Q000378:metabolism; D010084:Oxidation-Reduction; D010168:Palmitates / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D012265:Ribonucleotides / Q000494:pharmacology; D013268:Stimulation, Chemical
chemical_list: D002518:Ceramides; D005227:Fatty Acids; D020690:GTPase-Activating Proteins; D051275:Glucose Transporter Type 4; D005989:Glycerides; D007328:Insulin; D020738:Leptin; D010168:Palmitates; D010743:Phospholipids; D012265:Ribonucleotides; C495694:Slc2a4 protein, rat; C533101:TBC1D4 protein, rat; D000620:Aminoimidazole Carboxamide; D051058:Oncogene Protein v-akt; D000263:Adenylate Kinase; C031143:AICA ribonucleotide; D005947:Glucose