目的建立小鼠扩展胰岛素钳夹技术并观察其体内糖代谢的变化。方法分别采用[3-^3H]葡萄糖和2-脱氧-^3H葡萄糖(2-DOG)作为示踪剂,建立自由状态下小鼠正糖-高胰岛素钳夹技术,并观察其体内糖代谢的动态改变。结果在钳夹稳定状态,血糖稳定在基础水平(7.0±1.3mmol/L),在高血浆胰岛素水平下,血浆FFA明显抑制(从1.45±0.15到0.45±0.08mmol/L,P〈0.01)。葡萄糖输注率为46.7±4.7mg·kg^-1·min^-1,葡萄糖清除率为11.2±0.6mg·kg^-1·min^-1,肝糖产生被完全抑制,骨骼肌葡萄糖摄取率为20.9±2.5μmol·100g^-1·min^-1,而TG和TC浓度较钳夹前明显降低(P均〈0.01)。平均血糖变异系数为5.19%,平均GIR变异系数为9%。结论采用上述示踪剂建立的小鼠扩展胰岛素钳夹技术,能准确评价小鼠机体胰岛素敏感性,同时能动态反映体内糖代谢变化。
Objective To establish an euglycemic-hyperinsulinemic clamp technique in mice and evaluate the changes of glucose metabolism in vivo.Methods Hyperinsulinemic-euglycemic clamp technique with [3^-3H]glucose and 2-deoxy[3-^3H] glucose(2-DOG)as a tracer was establish in awake unstressed mice(insulin infusion rate:5mU·kg^-1·min^-1).And the glucose metabolism was determined in vivo.Results During clamp steady-state,blood glucose was 7.0 ± 1.3 mmol/L,plasma free fatty acids(FFA)were significantly suppressed(from 1.45±0.15 to 0.45±0.08mmol/L,P〈0.01),glucose infusion rates(GIR)was 46.7± 4.7mg·kg^-1 ·min^-1,glucose disappearance rate(GRd)was 11.2±0.6mg·kg^-1·min^-1,the glucose uptake rate in muscle(MGU)was 20.9 ± 2.5 μmol·100g^-1·min^-1,and hepatic glucose production(HGP)was completely inhibited,while triglyceride(TG)and cholesterol(TC)levels were remarkably decreased(all P〈0.01).The average coefficients of variation in blood glucose and GIR were 5% and 9%,respectively.Conclusions Hyperinsulinemic-euglycemic clamp technique with [3-^3H]glucose and 2-deoxy[3-^3H]glucose(2-DOG)as a tracer was established in awake unstressed mice for evaluating insulin sensitivity and glucose turnover.