| 14-3-3β蛋白介导胰高血糖素作用下糖异生的特点及机制 |
| 投稿时间：2016-07-21 修订日期：2016-07-25 点此下载全文 |
| 引用本文：冯丽帅,王倩倩,马旭,王建波,魏丽.14-3-3β蛋白介导胰高血糖素作用下糖异生的特点及机制[J].医学研究杂志,2017,46(3):36-38,47 |
| DOI：
10.11969/j.issn.1673-548X.2017.03.010 |
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| 基金项目:国家自然科学基金资助项目（面上项目）（81570778） |
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| 中文摘要:目的 分别观察胰高血糖素在空质粒转染组和14-3-3β蛋白过表达组对HepG2细胞糖异生的影响，并对14-3-3β蛋白介导此现象出现的可能原因初步探讨。方法 对转染空质粒或14-3-3β质粒低糖培养基培养的HepG2细胞分别在有无胰高血糖素作用下，进行培养基中葡萄糖产量的测定并观察糖异生关键酶蛋白表达量的变化；用免疫共沉淀的方法判定稳定转染胰高血糖素受体的293细胞在胰高血糖素对待下，14-3-3β蛋白与胰高血糖素受体结合的变化。结果 在未加及加入胰高血糖素对待下，14-3-3β转染组与空质粒对照组相比葡萄糖产量降低（P均<0.05）；糖异生关键酶表达量降低（P均<0.05）；免疫共沉淀显示胰高血糖素受体、14-3-3β蛋白与碳水化合物反应元件三者处于结合状态，且在胰高血糖素作用下14-3-3β蛋白与胰高血糖素受体结合减少而与碳水化合物反应元件结合增多（P均<0.05）。结论 过表达14-3-3β蛋白可起到抑制胰高血糖素的糖异生作用且这种现象发生的原因可能与其同碳水化合物反应元件间的相互作用及产生的后续效应有关。 |
| 中文关键词:胰高血糖素 胰高血糖素受体 14-3-3β蛋白 碳水化合物反应元件 |
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| Research on the Effect and Mechanism of 14-3-3 Beta Protein on Gluconeogenesis in the Action of Glucagon |
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| Abstract:Objective To observe the effect of glucagon on the expression of HepG2 cell and the expression of 14-3-3 protein in the empty plasmid group and the expression of 14-3-3 protein, and to explore the possible causes of the phenomenon. Methods The differences of glucose yield and the expression between empty plasmid or 14-3-3 beta plasmid transfected HepG2 cells to culture medium glucose yield of determination were obserred and the expression of key enzymes of gluconeogenesis glucagon action was obserred.with CO immunoprecipitation method,we determined the binding changes between glucagon receptor 14-3-3 protein and glucagon receptor under the treatment of glucagon. Results 14-3-3 beta plasmid transfected group was shown to yield less glucose compared with empty plasmid control group (P<0.05), and also less expression of gluconeogenesis enzymes (P<0.05). CO immunoprecipitation displayed the combination and also the alternative of bound state between glucagon receptor, 14-3-3 protein and carbohydrate response element under the treatment of glucagon (P<0.05). Conclusion Overexpression of 14-3-3 protein may inhibit the glucagon induced gluconeogenesis due to the characteristic of them in the combination with glucagon receptor and carbohydrate response element. |
| keywords:Glucagon Glucagon receptor 14-3-3 protein Carbohydrate response element |
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