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作 者:刘铭瑶[1] 王文飞[1] 侯玉婷 任桂萍[1] 吴云舟[1] 李德山[1]
机构地区:[1]东北农业大学生命科学学院,哈尔滨150030 [2]长春卫尔赛生物药业有限公司,长春130616
出 处:《东北农业大学学报》2016年第5期36-43,共8页Journal of Northeast Agricultural University
基 金:国家自然基金项目(J1210069/J0116);东北农业大学博士启动基金(2010RCB52)
摘 要:肝脏是代谢的中枢性器官,在糖脂代谢中扮演重要角色。FGF-21是近年来发现的一种治疗糖尿病新型药物,研究其对肝脏糖代谢影响及机制将为FGF-21成药性提供理论依据。以Hep G2细胞为肝细胞模型,探究FGF-21对Hep G2细胞葡萄糖吸收影响及作用机制。FGF-21处理Hep G2细胞,采用葡萄糖氧化酶-过氧化物酶(GOD-POD)法检测细胞对葡萄糖摄取情况,并检查胰岛素与FGF-21协同作用,蒽酮法检测细胞内糖原含量,半定量和实时荧光定量PCR检测FGF-21对葡萄糖转运蛋白(GLUTs)m RNA表达影响。结果表明,FGF-21可促进Hep G2细胞摄取葡萄糖,与胰岛素具有一定协同作用,增加糖原合成。半定量PCR结果显示在FGF-21作用下,仅GLUT1m RNA表达有所增加。实时荧光定量PCR检测FGF-21作用时间对GLUT1 m RNA表达量影响,发现FGF-21作用6 h时GLUT1 m RNA表达量倍数增加最大。说明FGF-21可通过增加GLUT1 m RNA表达促进Hep G2细胞消耗葡萄糖,参与糖原合成。Liver is a metabolic center, plays an important role in regulating glucose and lipid metabolism. FGF- 21 is an important candidate drug to treat diabetes; so to study the effect and mechanism of its effects on regulation of the glucose metabolism in liver is very important for drug research. We chose Hep G2 cells as a model to study the effects of FGF-21 on glucose uptake in liver and the mechanism of its action. After treated by FGF-21, the glucose uptake by the Hep G2 cells was detected by the method of glucose oxidizes · peroxides(GOD-POD); glycogen synthesis was examined by anthrone method, the synergy between insulin and FGF-21 was evaluated. With the purpose to study the mechanism of FGF- 21 on glucose uptake, the m RNA expression of GLUTs was detected by semiquantitative PCR and real- time PCR with specific primers. The results showed that FGF- 21 stimulated glucose uptake by the Hep G2 cells in a does-dependent manner and had a synergistic effect with insulin. FGF-21 could also increase glycogen synthesis in the Hep G2 cells, with the same action as insulin. The result of the semiquantative PCR showed that only GLUT1 m RNA was increased with FGF- 21 stimulation. FGF- 21 induced a significant increase of GLUT1 m RNA in the Hep G2 cells detected by real-time PCR. The fold of GLUT1 m RNA expression was raised most at 6 hours, more than 5-fold was increased. Thus, we concluded that FGF-21 stimulates glucose uptake and glycogen synthesis in Hep G2 cells, and enhanced glucose absorption of Hep G2 cells probably through GLUT1 activation.
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