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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

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【作者】 常宝成张宏郑凝赵伟张秋梅卫立君方佩华

【Author】 CHANG Bao-cheng,ZHANG Hong,ZHENG Ning,ZHAO Wei,ZHANG Qiu-mei,WEI Li-jun,FANG Pei-hua(1.Metabolic Disease Hospital;2.General Hospital,Tianjin Medical University,Tianjin 300070,China)

【机构】 天津医科大学代谢病医院天津医科大学总医院核医学科 天津300070天津300070

【摘要】 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(R IA)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。RPM I1640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、RPM I1640培养液中葡萄糖浓度及培养时间均与分离胰岛活性有关。

【Abstract】 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.

【基金】 天津市高等学校科技发展基金(20050204);卫生部笹川医学奖学金同学会科研启动基金(099)
  • 【文献出处】 基础医学与临床 ,Basic & Clinical Medicine , 编辑部邮箱 ,2006年09期
  • 【分类号】R-332
  • 【被引频次】9
  • 【下载频次】94
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