| Objective:To observe the effects of insulin intervention on lipid metabolism in L-6 myotubes with FFA-induced insulin resistance.Methods:L6 cell line was cultured and differentiated into myotubes in the 6-well board. After 12-16h of the starved condition in L-DMEM with 1% FFA-free BSA, L-DMEM with 0.4mM PA and 5%FFA-free BSA was added to the myotubes for 24h, the content of glucose in the supernatant was detected by using glucose oxidase method。Myotubes of PA were then replaced by L-DMEM with 10nmol insulin for 24h, The mRNA expression and protein levels of lipid metabolism related genes were measured by RT-PCR and western blot.Results:(1) The content of glucose in the supernatant of PA group was obviously higher than that of the control group, which suggested the insulin resistant model of skeletal muscle was successfully established.(2) The mRNA expression and the protein level of SREBP-lc in PA group was significantly up-regulated than the controls (p<0.05); the mRNA expression and the protein level of SREBP-1c were obviously down-regulated than the PA group after insulin intervention (p<0.05).(3) The mRNA expression and the protein level of IRS-1 of the PA group was significantly decreased(p<0.05), which were obviously increased after insluin intervention(p<0.05).(4) The mRNA expressions of TNFa in the control group, PA group and insulin group were not significantly changed (p>0.05).(5) Compared with the control group, the mRNA expression of stat3 were not significantly changed among the three groups(p>0.05).Conclusion:Insulin intervention may relieve insulin resistance potentially through inhibition of SREBP-lc in skeletal muscle. |
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