Arachidonate activation of protein kinase C may be involved in the stimulation of protein synthesis by insulin in L6 myoblasts.
Insulin stimulated protein synthesis in L6 myoblasts but did not increase the labelling of DAG or the release of phosphocholine from phosphatidylcholine. The DAG lipase inhibitor, RHC 80267, more than doubled the amount of label appearing in DAG but did not stimulate protein synthesis. Even in the presence of the DAG lipase inhibitor insulin failed to have any effect on DAG labelling, and conversely RHC 80267 did not modify the insulin-induced increase in protein synthesis. These results suggest that endogenous DAG production is not involved in the stimulation of protein synthesis by insulin. However, exogenous diacylglycerols (1-oleoyl-2-acetyl glycerol and 1-stearoyl-2-arachidonoyl glycerol) both stimulated protein synthesis in L6 myoblasts. The efficacy of the former (arachidonate-free) DAG suggested that their action was by activation of protein kinase C rather than by arachidonate release and prostaglandin formation. Ibuprofen, an inhibitor of cyclo-oxygenase failed to block the effects of insulin whereas a second cyclo-oxygenase inhibitor, indomethacin had only a partial inhibitory effect. The protein kinase C (PKC) inhibitor, RO-31-8220, totally blocked the effect of insulin. Since indomethacin is also recognised to inhibit phospholipase A2, the data suggests that insulin acts on protein synthesis in myoblasts by arachidonate activation of PKC.
Thompson
M G
MG
Rowett Research Institute, Bucksburn, Aberdeen, U.K.
Acamovic
F
F
Mackie
S C
SC
Morrison
K S
KS
Palmer
R M
RM
D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism*; D002460:Cell Line; D003512:Cyclohexanones / Q000494:pharmacology; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D004075:Diglycerides / Q000378:metabolism / Q000494:pharmacology; D004789:Enzyme Activation; D007211:Indoles / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology*; D008071:Lipoprotein Lipase / Q000037:antagonists & inhibitors; D009124:Muscle Proteins / Q000096:biosynthesis*; D009132:Muscles / Q000187:drug effects / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011744:Pyrimidinones / Q000494:pharmacology; D051381:Rats; D013844:Thiazoles / Q000494:pharmacology
D003512:Cyclohexanones; D016861:Cyclooxygenase Inhibitors; D004075:Diglycerides; D007211:Indoles; D007328:Insulin; D009124:Muscle Proteins; D011744:Pyrimidinones; D013844:Thiazoles; D016718:Arachidonic Acid; C075809:1-stearoyl-2-arachidonoylglycerol; C036535:1,6-bis(cyclohexyloximinocarbonyl)hexane; C038237:1-oleoyl-2-acetylglycerol; C047505:R 59022; D011493:Protein Kinase C; D008071:Lipoprotein Lipase; C064758:Ro 31-8220