Precise Assessment of Tissue-Specific Insulin Sensitivity: Complete Protocol for ¹³C-Glucose Clamp Technique

This protocol provides an improved hyperinsulinemic-euglycemic clamp technique. Through intravenous infusion of ¹³C-labeled glucose and insulin, it dynamically analyzes ¹³C-glucose metabolic flux using liquid chromatography-tandem mass spectrometry (LC-MS/MS) while stably maintaining blood glucose levels (euglycemia), thereby enabling quantitative assessment of insulin sensitivity in target tissues such as adipose and muscle.

Precise Tissue-Specific Analysis

• Targeting key metabolic tissues: Continuous infusion of insulin and ¹³C-glucose through jugular vein catheter, combined with steady-state glucose regulation technology (GIR regulation), directly quantifies glucose uptake rate (GDR) and hepatic gluconeogenesis inhibition capacity (HGP) in adipose and muscle tissues.
• Dynamic tracking of metabolic flow: utilizing LC-MS/MS technology to detect ¹³C-labeled metabolites, resolving the real-time flow of glucose in the body, breaking through the limitations of traditional clamps that only reflect systemic sensitivity.

Validated Model Applicability

Integrated a high-fat diet-induced insulin resistance model (C57BL/6J mice) and validated its reliability through multidimensional assessments including Akt phosphorylation levels and glucose tolerance tests (GTT/ITT/PTT).

• Integrated a high-fat diet-induced insulin resistance model (C57BL/6J mice) and validated its reliability through multidimensional assessments including Akt phosphorylation levels and glucose tolerance tests (GTT/ITT/PTT).

Applications

• Insulin resistance mechanism research
• Diabetes drug development
• Metabolic dynamics tracking

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