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(1)目的通过比较MIN6细胞损伤模型的不同建立方法,筛选最佳造模方法。(2)方法将生长状态良好的MIN6细胞分入96孔板,培养至对数生长期后分别以不同浓度棕榈酸(PA)、葡萄糖(GLU)、棕榈酸+葡萄糖(PA+GLU)、地塞米松(DXM)和过氧化氢(H_2O_2)干预12小时,CCK-8试剂盒检测细胞活力,计算IC50。设立对照组,以IC50浓度分别干预6小时后流式细胞仪检测细胞凋亡,ELISA法检测细胞胰岛素分泌。(3)结果细胞活力结果显示各造模剂均有良好的剂量依赖性;分别以IC50浓度干预后,细胞早期凋亡和晚期凋亡均比对照组有所增加,其中PA+GLU早期凋亡、胰岛素分泌量较高。(4)结论 PA+GLU干预MIN6细胞后早期凋亡比例增加和刺激胰岛素分泌增加比较接近体内情况,符合造模需求。
(1) Objective To compare the different establishment methods of MIN6 cell injury model and select the best method of modeling. (2) Methods MIN6 cells with good growth condition were divided into 96-well plates and cultured until the logarithmic growth phase were treated with different concentrations of palmitic acid (PA), glucose (GLU), palmitic acid + glucose (PA + GLU) DXM and H 2 O 2 for 12 hours, the cell viability was measured by CCK-8 kit, and the IC50 was calculated. The control group was set up. The apoptosis was detected by flow cytometry after 6 hours of IC50 concentration intervention, and the insulin secretion was detected by ELISA. (3) Results The results of cell viability showed that each model agent had a good dose-dependent manner. After IC50 concentration intervention, the early apoptosis and the late apoptosis were both increased compared with the control group, and the early apoptosis of PA + GLU , High insulin secretion. (4) Conclusion PA + GLU intervention MIN6 cells increased the proportion of early apoptosis and stimulated insulin secretion increased closer to the body, in line with modeling needs.