葡萄糖诱发小鼠胰岛β细胞氢离子流改变的测定

Proton flux of mouse pancreatic islet β cells induced by glucose

目的:实时记录C57BL/6J小鼠胰岛β细胞原位状态下的氢离子(H+)流,观察其在葡萄糖刺激下的改变,以了解其在胰岛素分泌过程中的作用。方法:取8周龄雄性C57BL/6J小鼠,急性分离胰岛,置于含10%小牛血清的RPMI1640培养基中孵育12h,应用非损伤微测技术检测20mmol/L葡萄糖对胰岛β细胞H+流的影响。结果:胰岛原位β细胞有基础性H+外流存在,检测48个胰岛的β细胞H+流,测得基础流速为(0.0205±0.0019)pmol/(cm2·s),葡萄糖刺激后的流速增加为(0.0519±0.0063)pmol/(cm2·s),差异显著(t=33.128,P<0.05)。结论:通过非损伤微测技术,首次实时记录到胰岛β细胞原位状态下的H+外流,并发现其在葡萄糖刺激下的流速增加,为进一步研究胰岛β细胞H+流在胰岛素分泌过程中的作用提供了实验基础,对于深入探讨糖尿病的发生机制具有重要作用。

Objective To investigate the role of proton flux on insulin secretion induced by glucose. Methods Pancreatic β cell islets were isolated from 8 weeks aged C57BL/6J male mouse and cultured in RPMI-1640 media for 12 hours. The non-invasive micro-test technique （NMT） was used to detect the status of proton flux initially and after stimulation by 20 mmol/L glucose. Results There was a basal proton effiux in the normal islet and the proton effiux was increased after glucose stimulation. The baseline velocity was 0.0205-＋0.0019 pmol/cm^2·s, and the velocity was significantly increased to 0.0519＋_0.0063 pmol/cm^2·s after the glucose exposure （t=33.128, P〈0.05）. Conclusions By NMT technique, this study observed the proton flux changes in mouse pancreatic islet β cells at baseline, and after glucose stimulation. This study elucidales the role of proton flux regulation in insulin secretion, and might help for the the mechanism research of diabetes.