INS-1E细胞葡萄糖毒性模型的建立

目的:建立胰岛细胞系INS-1E细胞的葡萄糖毒性模型。方法:将INS-1E细胞分别在不同葡萄糖浓度(5.5 mmol/L、16.7mmol/L、25 mmol/L、30 mmol/L)的1640完全培养基中培养不同时间(48 h、72 h、96 h、120 h),分别在不同时间点取细胞进行细胞功能检测,实时荧光定量PCR法检测胰岛素m RNA的表达,ELISA检测葡萄糖刺激的胰岛素的分泌。结果:与对照组相比,高糖浓度(5.5 mmol/L、16.7 mmol/L、25 mmol/L、30 mmol/L)培养基中培养48 h后,INS-1E细胞的胰岛素合成和分泌的功能均增加(P均0.05),随着培养基中葡萄糖浓度的升高以及培养时间的延长,INS-1E细胞胰岛素合成及分泌的功能逐渐下降,当在葡萄糖浓度为30 mmol/L的培养基中培养120 h后,胰岛素m RNA合成及葡萄糖刺激的胰岛素分泌均显著降低(P均0.01)。结论:INS-1E细胞在30 m M的葡萄糖中培养120 h形成稳定的葡萄糖毒性模型。

Establish Glucose ToxicityModel of INS-1E Cell

Objective:To establish glucose toxicity model of INS-1E cell.Methods:INS-1E cells were cultured in differentglucose concentration (5.5 mmol/L, 16.7 mmol/L, 25 mmol/L and 30 mmol/L) 1640 fully culture medium in different time (48 h, 72 h, 96h, 120 h). Cells were taken at different time points to detect cell function. Insulin mRNA was detected by fluorescent quantitative PCR.Insulin secretion was tested by ELISA.Results:Compared with the normal glucose control group, after 48 h cultured in high glucoseconcentrations (5.5 mmol/L, 16.7 mmol / L, 25 mmol / L, 30 mmol / L), insulin synthesis and secretion of INS-1E cells were increased(P<0.05). With increasing concentration of glucose in the medium and longer incubation time, insulin synthesis and secretion function ofINS-1E cells was gradually declined. When cells were cultured 120 h in the medium with 30 mmol/L glucose concentration, insulinmRNA synthesis and glucose-stimulated insulin secretion were significantly reduced (P<0.01).Conclusion:Glucose toxicity model canbe formed after INS-1E cell cultured 120 h in 30 mMglucose.