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| Effects of Duck (Anas platyrhynchos) SCD1 Gene Over-expression on Lipid Metabolism in Primary Hepatocytes |
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Abstract Stearoyl-CoA desaturase 1 (SCD1) is a rate-limiting enzyme for the synthesis of monounsaturated fatty acids in hepatocytes, and plays an important regulation role in the composition of fatty acids in the body. In order to explore the genetic regulation mechanism of SCD1 gene on the fat deposition of poultry, 21 days embryo duck (Anas platyrhynchos) hepatocytes were extracted by the method of type Ⅳ collagenaseand. The pEGFP-N3-SCD1 eukaryotic over-expression vector was constructed and transferred into the hepatocytes. The over-expression of SCD1 gene in duck hepatocytes was measured by HIS labeling assay kit to understand the effect of SCD1 on lipid metabolism. The experimental results showed that initial separation of hepatocytes was good, and the cells were basically covered with culture plates until 96 h, The recombinant plasmid pEGFP-N3-SCD1 and green fluorescence could over- expression stably in the primary hepatocytes of the ducks. The correlation between over-expression and lipid index showed that the expression of SCD1 was positively correlated with total cholesterol (TC) and low density lipoprotein (LDL) (P＜0.01), The content of high density lipoprotein (HDL) was significantly negative correlation (P＜0.01) but triglyceride (TG) (P＞0.05). The results showed that the recombinant plasmid pEGFP-N3-SCD1 could be successfully isolated and purified from the liver extraction by using type Ⅳ collagenase. Over-expression of SCD1 gene had a regulatory function for lipid metabolism of duck primary hepatocytes and it is useful for the future study about the genetic regulation mechanism of SCD1 on duck fat deposition.
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Received: 12 August 2017
Published: 14 February 2018
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