The Effect of Insulin Receptor Substrates 1 and 2 Knockdown on Porcine Hepatic Glucolipid Metabolism

Abstract

Objective: Through efficient shRNA interference to knockdown simultaneously IRS1 and IRS2 in porcine liver cells,and verify its effects on genes expression related to glucolipid metabolism,which may lay a solid foundation for producing the type2 diabetes mellitus model pigs. Method: First of all,cloning porcine IRS1 gene and the partial 3-untranslated region sequence of IRS2 gene by Overlap PCR and 3' RACE respectively. Then screening the most effective interference shRNA fragments of IRS1 and IRS2 by Real-time PCR. Finally, the glucolipid metabolism related genes experssion were detected in the porcine liver cells of both knocking down IRS1 and IRS2. Result: The knockdown of both IRS1 and IRS2 resulted in significant upregulation of gluconeogenic enzymes PEPCK and F-1,6-BP,as well as a decrease in Gck expression,at the same time,also led to the upregulation of lipogenic enzymes SREBP-1,Abcg8 and CYP7a1. Conclusion: The knockdown of both IRS1 and IRS2 may lead to an increasing level of blood glucose and also can cause a disorder of cholesterol metabolism. Therefore,IRS1 and IRS2 have important roles in the regulation of hepatic glucolipid metabolism.