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目的建立Caco-2细胞葡萄糖吸收模型,为考察药物对葡萄糖吸收的影响提供方法。方法 Caco-2细胞培养7,14,21 d后,分别加入25,50,75,100 mmol.L-1的葡萄糖溶液孵育5,10,15,20 min,采用高效液相色谱-质谱联用技术,以稳定同位素示踪法比较培养时间、葡萄糖浓度以及孵育时间对葡萄糖吸收的影响。结果 Caco-2细胞培养21 d后葡萄糖的吸收优于培养7 d和14 d的吸收,吸收量大,趋势稳定。此时用75 mmol.L-1的葡萄糖溶液孵育细胞10 min,葡萄糖吸收量与100 mmol.L-1比较差异无统计学意义,但高于25,50 mmol.L-1时的葡萄糖吸收量。结论本实验条件下建立的Caco-2细胞模型可作为研究葡萄糖吸收的模型。
Objective To establish a glucose uptake model of Caco-2 cells and provide a method to study the effect of drugs on glucose uptake. Methods Caco-2 cells were cultured for 7 days, 7 days, 21 days and 25, 75, 75, 100 mmol.L-1 glucose respectively. The cells were incubated for 5, 10, 15 and 20 min, and were separated by high performance liquid chromatography-mass spectrometry Stable isotope tracing method was used to compare the effect of incubation time, glucose concentration and incubation time on glucose uptake. Results Caco-2 cells cultured 21 d after the glucose absorption better than the 7 d and 14 d of absorption, absorption, the trend is stable. At this time, the cells were incubated with 75 mmol.L-1 glucose for 10 min, and there was no significant difference between the glucose uptake and 100 mmol.L-1, but higher than the glucose uptake at 25 and 50 mmol.L-1 . Conclusion The Caco-2 cell model established in this experiment can be used as a model for studying glucose uptake.