大鼠胰岛分离技术的建立及胰岛活性的影响因素

Isolating method of rat pancreatic islets and affect factors for its activity

目的 建立稳定有效的大鼠胰岛分离方法,探讨影响胰岛活性的因素.方法 通过胆管注射胶原酶方法提取大鼠胰岛,通过水浴法比较不同条件下葡萄糖刺激的胰岛素分泌,用放免法（RIA）测定胰岛素.结果 牛血清白蛋白（BSA）能明显提高葡萄糖刺激的胰岛素分泌水平,长时间培养（＞7 d）的胰岛,其葡萄糖刺激的胰岛素分泌下降约25%,而新鲜分离的胰岛和经1～5 d培养的胰岛未见明显差异.RPMI1640培养液中葡萄糖浓度11.1 mmol/L组,其胰岛素分泌明显高于5.5 mmol/L和25 mmol/L这2组.结论 BSA、RPMI1640培养液中葡萄糖浓度及培养时间均与分离胰岛活性有关.

Objective To assess the factors which affect the activity of isolated islets, and to construct a steady and effective isolation method of rat pancreatic islets. Methods Pancreatic islets were isolated by collagenase method. The glucose-stimulation of insulin secretion（GSIS） in different conditions was performed by batch incubation method and measured by RIA. Results GSIS was improved by BSA; There was a evident decrease of insulin secretion in cultured islets of 7 days, however, no difference was observed between the freshly isolated islets group and the 1-5days cultured islets groups. Compared to the groups of 5.5 mmoL/L and 25 mmoL/L glucose in the culture medium, 11.1 mmoL/L glucose group stimulated higher insulin release. Conclusion BSA, glucose concentration of RPMI1640 medium and the culture period are related to the activity of isolated islets.