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| Effects of Silencing ACSL1 Gene by siRNA on the Synthesis of Unsaturated Fatty Acids in Adipocytes of Qinchuan Beef Cattle (Bos taurus) |
| TIAN Hong-Shan1, SU Xiao-Tong2, ZHAO Zhi-Dong1,*, HAN Xiang-Min1, ZAN Lin-Sen2,3,*, HU Jiang1, LUO Yu-Zhu1, WANG Ji-Qing1, BAI Yan-Bin1 |
1 College of Animal Science and Technology, Gansu Agricultural University/Gansu Key Laboratory of Herbivorous Animal Biotechnology, Lanzhou 730070, China;
2 College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China;
3 National Beef Cattle Improvement Center, Yangling 712100, China |
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Abstract Long-chain acyl-CoA synthetase 1 (ACSL1) is a member of the acyl activating enzyme family, which plays an important role in the activation, transport, degradation and synthesis of fatty acids in vivo. In this study, the adipocytes of Qinchuan beef cattle (Bos taurus) were used as the research object. qRT-PCR and small interfering RNA (siRNA) interference were performed to explore the mechanism of ACSL1 gene on the synthesis of unsaturated fatty acid. Firstly, the temporal expression of ACSL1 gene and unsaturated fatty acid synthesis related genes, such as fatty acid synthase (FASN), stearoyl-CoA desaturease 1 (SCD1), peroxisome proliferator activated receptor gamma (PPARγ), fatty acid desaturase 1 (FADS1) and fatty acid binding protein 3 (FABP3) during adipocyte differentiation were detected, and it was found that the expression levels of ACSL1, FADS1, SCD1, FASN and PPARγ showed a trend of first increasing and then decreasing, and peaked at 4 d, while FABP3 had no significant changes during the differentiation of bovine adipocytes. Then, siRNA targeting the ACSL1 gene was transfected into bovine adipocytes, and found that the mRNA level of ACSL1 was down-regulated by more than 70% (P<0.01), and the protein level was also significantly down-regulated. After silencing the ACSL1 gene, the mRNA level of FABP3 and PPARγ were down-regulated, and FADS1 and FASN were up-regulated, while SCD1 showed a trend of increasing first and then decreasing. Furthermore, interfering ACSL1 gene expression significantly reduced the ratio of C18∶0 (P<0.05), so that the content of saturated fatty acid (SFA) in adipocytes was reduced. Meanwhile, the ratio of C16∶1, C18∶1, C18∶2, arachidonic acid (AA) and eicosapentaenoic acid (EPA) decreased significantly (P<0.01), so that the content of saturated fatty acid (SFA) and polyunsaturated fatty acids (PUFA) in adipocytes was decreased. In addition, Bodipy staining showed that silencing the ACSL1 gene could reduce the formation of lipid droplets in bovine adipocytes. In summary, the ACSL1 gene played an important role in regulating the expression of unsaturated fatty acid synthesis-related genes, fatty acid composition and lipid droplet formation, and provides a theoretical basis for further elucidating the molecular mechanism of unsaturated fatty acid formation in bovine adipocytes.
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Received: 14 February 2020
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Corresponding Authors:
* zhaozd@gsau.edu.cn; zanlinsen@163.com
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