胰岛新生相关蛋白改善胰岛功能的研究
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国家自然科学基金面上项目(30671010;30971405)


Impacts of islet neogenesis-associated protein on islet function
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    摘要:

    目的: 观察胰岛新生相关蛋白(islet neogenesis-associated protein,INGAP)与胰岛体外共培养对其凋亡-基因表达以及胰岛素释放功能的影响-方法:分离提取8周龄雄性Wistar大鼠胰岛,培养于添加(共培养组)或未添加(对照组)INGAP(10 μg/ml)的RPMI 1640完全培养液中-24 h后收集INGAP共培养组和对照组胰岛,进行吖啶橙-碘化丙啶染色(AO-PI)-葡萄糖刺激的胰岛素释放试验及RT-PCR检测凋亡相关B细胞淋巴瘤/白血病-2(Bcl-2)基因和蛋白激酶B(Akt)基因mRNA的表达水平-结果:①INGAP共培养组的胰岛在高糖刺激下的胰岛素释放量高于对照组[(185.00±20.01) μU/ml vs. (58.67±17.03) μU/ml],并且刺激指数亦明显增高(5.10±2.35 vs. 1.92±0.42)-②INGAP共培养组与对照组相比,胰岛Bcl-2基因 (0.607±0.217 vs. 0.503±0.209)和Akt基因 (1.115±0.189 vs. 0.935±0.162) 的mRNA表达水平较明显增加-③共培养组的胰岛细胞存活率明显高于对照组(2 685.45±5.08 vs. 2 108.65±4.02)-结论:INGAP与胰岛体外共培养可以减少胰岛的凋亡,维持胰岛的功能-

    Abstract:

    Objective: To explore the effect of islet neogenesis-associated protein(INGAP) on islet function, apoptosis, gene expression and insulin secretion in vitro. Methods:Islets isolated from male adult Wistar rats were cultured in RPMI 1640 with or without INGAP(10 μg/ml). After 24 h, cultured islets were collected, the degree of apoptosis, glucose-stimulated insulin secretion were measured and the expression of genes Bcl-2 and Akt were detected by RT-PCR assay. Results: ① Glucose stimulated insulin secretion(GSIS) stimulated by 16.7 mmol/L glucose was increased in islets co-cultured with INGAP of compared with islets cultured without INGAP [(185.00±20.01) μU/ml vs.(58.67±17.03) μU/ml]; the stimulating index of islets of INGAP group(5.10±2.35) was higher than that of control (no INGAP group:1.92±0.42). ②Gene expression of Bcl-2 and Akt in INGAP group were increased compared with those of control group(0.607±0.217 vs. 0.503±0.209;1.115±0.189 vs. 0.935±0.162).③Islets survival in co-cultured pancreatic islet group(2 685.45±5.08)was higher than that of control group(2 108.65±4.02). Conclusion:INGAP can reduce the apoptosis of pancreatic islets and maintain islet function after co-culture in vitro.

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陈 恒,王 云,于晓竹,许馨予,徐宽枫,杨 涛,周红文.胰岛新生相关蛋白改善胰岛功能的研究[J].南京医科大学学报(自然科学版),2010,(11):1564-1567

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  • 收稿日期:2010-07-06
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