|
|
Effect of SLC27A6 Expression Inhibition on Lipid Metabolism in Bovine (Bos taurus) Mammary Epithelial Cells |
SHEN Zi-Liang, JIANG Hui, CHEN Dai-Jie, HAN Zi-Yin, MAO Yong-Jiang, LI Ming-Xun, YANG Zhang-Ping, ZHANG Hui-Min* |
College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China |
|
|
Abstract Fatty acid compositions are crucial indicators of milk quality in dairy cow. Long chain fatty acid (LCFA), especially polyunsaturated fatty acids (PUFA) are beneficial in reducing the risk of cardiovascular disease, cancer, and type 2 diabetes in humans (Homo sapiens). However, the metabolic mechanism of LCFA in mammary gland is still unknown. Solute carrier family 27 member 6 (SLC27A6) is a transmembrane protein that could enhance the uptake of LCFA into cells, thus regulating downstream fatty acid (FA) metabolic pathways. But the relationship between SLC27A6 and fatty acid metabolic genes is still unclear. In this study, SLC27A6 knockdown in bovine (Bos taurus) mammary epithelial cells (BMECs) was accomplished by RNA interference to further assess the function of SLC27A6. The relative mRNA expression level of genes related to lipid metabolism after SLC27A6 knockdown in BMECs was analyzed by real-time quantitative PCR, and the protein level of SLC27A6 was analyzed by Western blot. The effect of SLC27A6 knockdown on triglyceride (TG) content and FA composition in BMECs were determined.The result showed that the mRNA and protein levels of SLC27A6 were decreased in SLC27A6-Bos-915 group compared with the control group. And knockdown of SLC27A6 dramatically downregulated the mRNA abundance of genes associated with FA activation long-chain acyl-CoA synthetase 4 (ACSL4), fatty acid transporter CD36 and oxidation carnitine palmitoyl transferase 1A (CPT1A), while abundance of genes associated with transcription regulation peroxisome proliferative activated receptor gamma (PPARG), fatty acid binding protein 3 (FABP3), and fatty acid desaturase 2 (FADS2) were upregulated. In addition, SLC27A6 was silenced, the intracellular content of TG and the percentage of oleic acid (C18∶1cis9) and arachidonic acid (C20∶4cis5,8,11,14) were higher, whereas that of palmitic acid (C16∶0) and stearic acid (C18∶0) were lower. Overall, these results could provide strong support for a central role of SLC27A6 in the regulation of lipid metabolism in BMECs, and have an important guiding significance to improving milk quality by breeding programs.
|
Received: 30 December 2020
|
|
Corresponding Authors:
* minmin-911@163.com
|
|
|
|
[1] 石恒波. 2014. PPARG基因对奶山羊乳腺脂质代谢的调控作用研究[D]. 博士学位论文, 西北农林科技大学, 导师: 罗军, pp. 9-10. (Shi H B.2014. The regulatory role of PPARG gene in fatty acid metabolism of goat mammary gland[D]. Thesis for D.C., Northwest A&F University, Supervisor: Luo J, pp. 9-10.) [2] 魏婷, 张永煜, 张庆华, 等. 2013. 肉毒碱棕榈酰基转移酶1A的研究进展[J]. 生命科学, 25(6): 614-620. (Wei T, Zhang Y Y, Zhang Q H, et al.2013. Research advances of carnitine palmitoyltransferase 1A[J]. Chinese Bulletin of Life Sciences, 25(6): 614-620.) [3] 许会芬. 2016. SREBP-1基因对山羊乳腺上皮细胞脂质代谢的调控作用研究[D]. 博士学位论文, 西北农林科技大学, 导师: 罗军, pp 19-20. (Xu H F.2012. The regulatory function of SREBP-1 gene on fatty acid metabolism in goat mammary gland epithelial cells[D]. Thesis for D.C., Northwest A&F University, Supervisor: Luo J, pp. 19-20.) [4] 绪欣. 2018. miR-145靶向FSCN1调控奶牛金葡菌型乳腺炎的分子功能初步探讨[D]. 硕士学位论文, 扬州大学, 导师: 杨章平, pp.12-28. (Xu X.2018. The preliminary discussion of molecular function of mir-145targeting FSCNl to regulate the Staphylococcus aureus mastitis of dairy cows[D]. Thesis for M.S., Yangzhou University, Supervisor: Yang Z P, pp. 12-28.) [5] Anderson C M, Stahl A.2013. SLC27 fatty acid transport proteins[J]. Molecular Aspects of Medicine, 34(2-3): 516-528. [6] Bionaz M, Loor J J.2008a. ACSL1, AGPAT6, FABP3, LPIN1, and SLC27A6 are the most abundant isoforms in bovine mammary tissue and their expression is affected by stage of lactation[J]. The Journal of Nutrition, 138(6): 1019-1024. [7] Bionaz M, Loor J J.2008b. Gene networks driving bovine milk fat synthesis during the lactation cycle[J]. BMC Genomics, 9(1): 366. [8] Blahova Z T N, Harvey M, Psenicka M, et al.2020. Assessment of fatty acid desaturase (FADS2) structure-function properties in fish in the context of environmental adaptations and as a target for genetic engineering[J]. Biomolecules, 10(2): 206. [9] Briant L J B, Dodd M S, Chibalina M V, et al.2018. CPT1A-dependent long-chain fatty acid oxidation contributes to maintaining glucagon secretion from pancreatic islets[J]. Cell Reports, 23(11): 3300-3311. [10] Chen Z, Chu S, Liang Y, et al.2021. miR-497 regulates fatty acid synthesis via LATS2 in bovine mammary epithelial cells[J]. Food & Function, 11(10): 8625-8636. [11] Chiu H C, Kovacs A, Blanton R M, et al.2005. Transgenic expression of fatty acid transport protein 1 in the heart causes lipotoxic cardiomyopathy[J]. Circulation Research, 96(2): 225-233. [12] Dixit S P, Sivalingam J, Tyagi A K, et al.2015. Association of novel SNPs in the candidate genes affecting caprine milk fatty acids related to human health[J]. Meta Gene, 4: 45-56. [13] Doege H, Baillie R A, Ortegon A M, et al.2006. Targeted deletion of FATP5 reveals multiple functions in liver metabolism: Alterations in hepatic lipid homeostasis[J]. Gastroenterology, 130(4): 1245-1258. [14] Doege H, Stahl A.2006. Protein-mediated fatty acid uptake: Novel insights from in vivo models[J]. Physiology, 21: 259-268. [15] Harvatine K J, Boisclair Y R, Bauman D E, et al.2009. Recent advances in the regulation of milk fat synthesis[J]. Animal, 3(1): 40-54. [16] Jia Z Z, Pei Z T, Maiguel D,et al.2007. The fatty acid transport protein (FATP) family: Very long chain acyl-CoA synthetases or solute carriers?[J]. Journal of Molecular Neuroscience, 33(1): 25-31. [17] Lee H, Park W J.2014. Unsaturated fatty acids, desaturases, and human health[J]. Journal of Medicinal Food, 17(2): 189-197. [18] Livak K J, Schmittgen T D.2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method[J]. Methods, 25(4): 402-408. [19] Lopes-Coelho F, André S, Félix A, et al.2018. Breast cancer metabolic cross-talk: Fibroblasts are hubs and breast cancer cells are gatherers of lipids[J]. Molecular and Cellular Endocrinology, 462(Pt B): 93-106. [20] Moody L, Xu G B, Chen H, et al.2019. Epigenetic regulation of carnitine palmitoyltransferase 1 (CPT1A) by high fat diet[J]. Biochimica Et Biophysica Acta-Gene Regulatory Mechanisms, 1862(2): 141-152. [21] Nafikov R A, Schoonmaker J P, Korn K T, et al.2013. Association of polymorphisms in solute carrier family 27, isoform A6 (SLC27A6) and fatty acid binding protein-3 and fatty acid binding protein-4 (FABP3 and FABP4) with fatty acid composition of bovine milk[J]. Journal of Dairy Science, 96(9): 6007-6021. [22] Palombo V, Loor J J, D'Andreaet M, et al.2018. Transcriptional profiling of swine mammary gland during the transition from colostrogenesis to lactogenesis using RNA sequencing[J]. BMC Genomics, 19(1): 322. [23] Pohl J, Ring A, Hermann T, et al.2004. Role of FATP in parenchymal cell fatty acid uptake[J]. Biochimica Et Biophysica Acta-Molecular and Cell Biology of Lipids, 1686(1-2): 1-6. [24] Qiu F F, Xie L, Ma J E, et al.2017. Lower Expression of SLC27A1 enhances intramuscular fat deposition in chicken via down-regulated fatty acid oxidation mediated by CPT1A[J]. Frontiers in Physiology, 8: 449. [25] Shi H B, Zhao W, Zhang C, et al.2016. Transcriptome-wide analysis reveals the role of PPARG controlling the lipid metabolism in goat mammary epithelial cells epithelial cells[J]. PPAR Research, 9195680. [26] Stahl A.2004. A current review of fatty acid transport proteins (SLC27)[J]. Pflugers Arch-Eur J Physiol, 447(5): 722-727. [27] Stefanovic-Racic M, Perdomo G, Mantell B S, et al.2008. A moderate increase in carnitine palmitoyltransferase 1a activity is sufficient to substantially reduce hepatic triglyceride levels[J]. American Journal of Physiology-Endocrinology and Metabolism, 294(5): E969-E977. [28] Vargas-Bello-Perez E, Zhao W, Bionaz M, et al.2019. Nutrigenomic effect of saturated and unsaturated long chain fatty acids on lipid-related genes in goat mammary epithelial cells: What is the role of PPAR gamma?[J]. Veterinary Sciences, 6(2): 54. [29] Wang Y, Li C, Wang Q Y, et al.2015. Danqi pill regulates lipid metabolism disorder induced by myocardial ischemia through FATP-CPTI pathway[J]. BMC Complementary and Alternative Medicine, 15: 28. [30] Yen M C, Chou S K, Kan J Y, et al.2019. New insight on solute carrier family 27 member 6 (SLC27A6) in tumoral and non-tumoral breast cells[J]. International Journal of Biological Sciences, 16(3): 366-375. |
|
|
|