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Updated: Jul 2, 2025
Glucose Uptake Measurement and Response to Insulin Stimulation in In Vitro Cultured Human Primary Myotubes
Published on: June 25, 2017
GT-11 impairs insulin signaling through modulation of sphingolipid metabolism in C2C12 myotubes
Javokhir Rustamov1, Yoon-Seok Roh1, Jin Tae Hong1
1College of Pharmacy, Chungbuk National University, Osongsaengmyeong 1-ro, Heungduk-gu, Cheongju, Republic of Korea.
Aims:
Sphingolipids are involved in the regulation of insulin signaling, which is linked to the development of insulin resistance, leading to diabetes mellitus. We aimed to study whether modulation of sphingolipid levels by GT-11 may regulate insulin signaling in C2C12 myotubes.
Main Methods:
We investigated the effects of sphingolipid metabolism on Akt phosphorylation and glucose uptake using C2C12 myotubes. Either GT-11, an inhibitor of dihydroceramide desaturase 1 and S1P lyase, or siRNA targeting Sgpl1, the gene encoding the enzyme, was employed to determine the effect of sphingolipid metabolism modulation on insulin signaling. Western blotting and glucose uptake assays were used to evaluate the effect of treatments on insulin signaling. Sphingolipid metabolites were analyzed by high performance liquid chromatography (HPLC).
Key Findings:
Treatment with GT-11 resulted in decreased Akt phosphorylation and reduced glucose uptake. Silencing the Sgpl1 gene, which encodes S1P lyase, mimicked these findings, suggesting the potential for regulating insulin signaling through S1P lyase modulation. GT-11 modulated sphingolipid metabolism, inducing the accumulation of sphingolipids. Using PF-543 and ARN14974 to inhibit sphingosine kinases and acid ceramidase, respectively, we identified a significant interplay between sphingosine, S1P lyase, and insulin signaling. Treatment with either exogenous sphingosine or palmitic acid inhibited Akt phosphorylation, and reduced S1P lyase activity.
Significance:
Our findings highlight the importance of close relationship between sphingolipid metabolism and insulin signaling in C2C12 myotubes, pointing to its potential therapeutic relevance for diabetes mellitus.
Insights
Modulating sphingolipid metabolism with GT-11 or Sgpl1 gene silencing impacts insulin signaling and glucose uptake in C2C12 myotubes. These findings suggest therapeutic potential for diabetes mellitus by targeting sphingolipid pathways.
Area of Science:
- Biochemistry
- Cell Biology
- Metabolic Research
Background:
- Sphingolipids play a crucial role in regulating insulin signaling pathways.
- Dysregulation of sphingolipid metabolism is implicated in insulin resistance and the development of diabetes mellitus.
Purpose of the Study:
- To investigate the effect of modulating sphingolipid levels using GT-11 on insulin signaling in C2C12 myotubes.
- To explore the potential of targeting sphingolipid metabolism for therapeutic interventions in diabetes mellitus.
Main Methods:
- C2C12 myotubes were treated with GT-11 (a dihydroceramide desaturase 1 and S1P lyase inhibitor) or siRNA targeting Sgpl1.
- Western blotting and glucose uptake assays assessed insulin signaling.
- High-performance liquid chromatography (HPLC) analyzed sphingolipid metabolites.
Main Results:
- GT-11 treatment decreased Akt phosphorylation and glucose uptake.
- Sgpl1 gene silencing mimicked these effects, indicating S1P lyase modulation's role.
- Interplay between sphingosine, S1P lyase, and insulin signaling was observed, with exogenous sphingosine or palmitic acid inhibiting Akt phosphorylation and reducing S1P lyase activity.
Conclusions:
- Sphingolipid metabolism is closely linked to insulin signaling in C2C12 myotubes.
- Modulating sphingolipid metabolism, particularly S1P lyase, shows therapeutic promise for diabetes mellitus.
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