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| The Effect of Insulin Receptor Substrates 1 and 2 Knockdown on Porcine Hepatic Glucolipid Metabolism |
| HUANG Tian-qing1, KONG Qing-ran1, LI Yan1, YU Miao2, LIU Zhong-hua1 |
1. Laboratory of Embryo Engineering, Department of Life Science, Northeast Agricultural University, Harbin 150030, China;
2. Harbin Power Vocational Technology College, Harbin 150030, China |
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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.
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Received: 02 January 2014
Published: 25 April 2014
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