硒对氧化损伤大鼠肝细胞糖代谢关键酶影响

Effects of selenium on glycometabolism key enzymes in hepatocytes of rats with oxidative damage

目的研究硒对氧化损伤大鼠肝细胞糖代谢关键酶表达的影响,并初步探讨其作用机制。方法采用0.1 mmol/L的H2O2建立氧化损伤细胞模型,并施加不同剂量硒干预,通过实时定量PCR技术检测葡萄糖激酶（glu-cok inse,GK）、糖原合成酶（glycogen synthase,GS）和蛋白激酶B（protein kinase B,PKB/Akt）的mRNA表达,并采用蛋白免疫印迹（W estern-b lot）方法检测Akt的蛋白表达水平。结果补硒各组GK的mRNA表达量为（9.692~16.588）×10-6,均高于H2O2损伤组（P〈0.05）;高剂量补硒组GS和Akt的mRNA表达量分别为57.618×10-6和0.2398×10-3,均高于H2O2损伤组（P〈0.05）;补硒各组Akt的蛋白表达量为（0.3343~0.4346）×10-3,均高于H2O2损伤组（P〈0.05）。结论补硒可以在一定程度上改善氧化损伤对肝细胞糖代谢关键酶GK、GS的影响,其机制可能与上调胰岛素信号传导通路的关键信号分子Akt的表达有关。

Objective To study effects of selenium on glycometabolism key enzymes and its mechanism in hepatocytes of rats with oxidative damage.Methods Rat hepatocytes were exposed to 0.1 mM H2O2 and then were incubated with different dosages of selenium for 24 hours.The mRNA expression of glucokine（GK）,glycogensymthase（GS）,and protein kinase B（PKB/Akt） were detected with real-time PCR.The protein expression of PKB/Akt was measured with Western blot.Results The mRNA expression of GK in selenium groups was 9.692-16.588×10-6,higher than that of in H2O2 oxidative model group（P0.05）.The mRNA expression of GS and Akt in high-dose selenium were 57.618 and 0.2398×10-3,higher than those of in H2O2 oxidative model group（P0.05）.The expression of protein in selenium groups was 0.3343-0.4346×10-3,higher than that of in H2O2 oxidative model group（P0.05）.Conclusion Selenium could improve the glucometabolism key enzymes GK and GS in rat hepatocytes with oxidative damage.The mechanism of the effect may relate to that selenium could up-regulate the expression of Akt which is the key molecule in insulin signal pathway.