FGF-21促进HepG2细胞摄取葡萄糖研究

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。

Study on FGF-21 regulates glucose uptake in HepG2 cells

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 HepG2 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 HepG2 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 mRNA 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 HepG2 cells in a does-dependent manner and had a synergistic effect with insulin. FGF-21 could also increase glycogen synthesis in the HepG2 cells, with the same action as insulin. The result of the semiquantative PCR showed that only GLUT1 mRNA was increased with FGF-21 stimulation. FGF-21 induced a significant increase of GLUT1 mRNA in the HepG2 cells detected by real-time PCR. The fold of GLUT1 mRNA 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 HepG2 cells, and enhanced glucose absorption of HepG2 cells probably through GLUT1 activation.