葡萄糖对鹅原代肝细胞GPX1和SOD1基因表达及酶活力的影响

Effect of Glucose on Gene Expression and Enzyme Activity of GPX1 and SOD1 in Goose Primary Hepatocytes

试验旨在研究葡萄糖对鹅肝细胞氧化应激的影响。通过RT-PCR方法扩增鹅肝脏谷胱甘肽过氧化物酶1(GPX1)和超氧化物歧化酶1(SOD1)基因的部分序列,分离、培养天府肉鹅原代肝细胞,并在添加不同浓度(0(对照组)、5、25、35mmol/L)葡萄糖培养基中培养48h,检测GPX和SOD活力及GPX1和SOD1mRNA表达水平。结果表明,鹅GPX1与斑胸草雀的同源性最高,SOD1与原鸡的同源性最高;与对照组相比,5mmol/L葡萄糖对GPX活力的影响不显著(P>0.05),25和35mmol/L葡萄糖均显著降低GPX活力(P<0.05);5mmol/L葡萄糖显著增加SOD活力(P<0.05),25和35mmol/L葡萄糖对SOD活力均无显著影响(P>0.05);5和25mmol/L葡萄糖对GPX1和SOD1mRNA表达均无显著影响(P>0.05),高浓度葡萄糖(35mmol/L)可显著增加GPX1和SOD1基因的表达水平(P<0.05)。提示,高浓度葡萄糖(35mmol/L)能引起鹅肝细胞内氧化应激的产生。

In order to investigate the effect of glucose on oxidative stress in cultured goose primary hepatocytes, partial gene sequences of GPX1 and SOD1 genes were cloned, and the goose primary hepatocytes were treated with different concentrations （0（control group）, 5, 25 and 35 mmol/L） of glucose for 48 h, and then enzyme activities of GPX and SOD and mRNA levels of GPX1 and SOD1 were examined. The results showed that goose GPX1 gene sequence had the highly similarity with Zebra Finch, and SOD1 had the highly similarity with Red Jungle fowl. Compared with the control group, 5 mmol/L glucose had no significant effect on GPX enzyme activity （P〉0.05）, but 25 and 35 mmol/L glucose could significantly decrease GPX enzyme activity （P〈0.05）. 5 mmol/L glucose could significantly increase SOD enzyme activity （P〈0.05）, but 25 and 35 mmol/L glu- cose had no significant effects on SOD enzyme activity（P〉0.05）. 35 mmol/L glucose could significantly increase mRNA expres- sion of GPX1 and SOD1 genes （P〈0.05）, but 5 and 25 mmol/L glucose had no significant effects on the two genes（P〉0. 05）. In conclusion, the high concentration glucose （35 mmol/L） could induce intracellular oxidative stress in goose primary hepatocytes.