Overexpression of carnitine palmitoyltransferase-1 in skeletal muscle is sufficient to enhance fatty acid oxidation and improve high-fat diet-induced insulin resistance.
OBJECTIVE
Skeletal muscle insulin resistance is associated with lipid accumulation, but whether insulin resistance is due to reduced or enhanced flux of long-chain fatty acids into the mitochondria is both controversial and unclear. We hypothesized that skeletal muscle-specific overexpression of the muscle isoform of carnitine palmitoyltransferase 1 (CPT1), the enzyme that controls the entry of long-chain fatty acyl CoA into mitochondria, would enhance rates of fatty acid oxidation and improve insulin action in muscle in high-fat diet insulin-resistant rats.


METHODS
Rats were fed a standard (chow) or high-fat diet for 4 weeks. After 3 weeks, in vivo electrotransfer was used to overexpress the muscle isoform of CPT1 in the distal hindlimb muscles (tibialis anterior and extensor digitorum longus [EDL]). Skeletal muscle insulin action was examined in vivo during a hyperinsulinemic-euglycemic clamp.


RESULTS
In vivo electrotransfer produced a physiologically relevant increase of approximately 20% in enzyme activity; and although the high-fat diet produced insulin resistance in the sham-treated muscle, insulin action was improved in the CPT1-overexpressing muscle. This improvement was associated with a reduction in triacylglycerol content, the membrane-to-cytosolic ratio of diacylglycerol, and protein kinase C theta activity. Importantly, overexpression of CPT1 did not affect markers of mitochondrial capacity or function, nor did it alter skeletal muscle acylcarnitine profiles irrespective of diet.


CONCLUSIONS
Our data provide clear evidence that a physiological increase in the capacity of long-chain fatty acyl CoA entry into mitochondria is sufficient to ameliorate lipid-induced insulin resistance in muscle.
Bruce
Clinton R
CR
Cellular and Molecular Metabolism Laboratory, Baker IDI Heart and Diabetes Institute, Melbourne, Victoria, Australia. clinton.bruce@baker.edu.au
Hoy
Andrew J
AJ
Turner
Nigel
N
Watt
Matthew J
MJ
Allen
Tamara L
TL
Carpenter
Kevin
K
Cooney
Gregory J
GJ
Febbraio
Mark A
MA
Kraegen
Edward W
EW
D000818:Animals; D002334:Carnitine O-Palmitoyltransferase / Q000235:genetics*; D004041:Dietary Fats / Q000009:adverse effects*; D018274:Electroporation; D005227:Fatty Acids / Q000378:metabolism*; D015309:Glucose Clamp Technique; D006234:Handling, Psychological; D006801:Humans; D006946:Hyperinsulinism; D007333:Insulin Resistance / Q000235:genetics*; D008297:Male; D008931:Mitochondria, Muscle / Q000201:enzymology; D018485:Muscle Fibers, Skeletal / Q000201:enzymology; D018482:Muscle, Skeletal / Q000201:enzymology*; D019308:Palmitic Acid / Q000378:metabolism; D010957:Plasmids; D016133:Polymerase Chain Reaction; D051381:Rats; D017208:Rats, Wistar; D015290:Second Messenger Systems / Q000502:physiology
D004041:Dietary Fats; D005227:Fatty Acids; D019308:Palmitic Acid; D002334:Carnitine O-Palmitoyltransferase