Metabolic Transition of Milk Triacylglycerol Synthesis in Response to Varying Levels of Three 18-Carbon Fatty Acids in Porcine Mammary Epithelial Cells

• Main Authors: Yantao Lv, Fang Chen, Shihai Zhang, Jun Chen, Yinzhi Zhang, Min Tian, Wutai Guan
• Format: Article
• Language: English
• Published: MDPI AG 2021-01-01
• Series: International Journal of Molecular Sciences
• Subjects: 18-carbon fatty acids; porcine mammary epithelial cells; milk fat biosynthesis; lipogenic genes
• Online Access: https://www.mdpi.com/1422-0067/22/3/1294

This study aimed to examine the effects of increasing levels of three 18-carbon fatty acids (stearate, oleate and linoleate) on mammary lipogenesis, and to evaluate their effects on the milk lipogenic pathway in porcine mammary epithelial cells (pMECs). We found that increasing the three of 18-carbon fatty acids enhanced the cellular lipid synthesis in a dose-dependent manner, as reflected by the increased (triacylglycerol) TAG content and cytosolic lipid droplets in pMECs. The increased lipid synthesis by the three 18-carbon fatty acids was probably caused by the up-regulated expression of major genes associated with milk fat biosynthesis, including <i>CD36</i> (long chain fatty acid uptake); <i>GPAM</i>, <i>AGPAT6</i>, <i>DGAT1</i> (TAG synthesis); <i>PLIN2</i> (lipid droplet formation); and <i>PPARγ</i> (regulation of transcription). Western blot analysis of CD36, DGAT1 and PPARγ proteins confirmed this increase with the increasing incubation of 18-carbon fatty acids. Interestingly, the mRNA expressions of <i>ACSL3</i> and <i>FABP3</i> (fatty acids intracellular activation and transport) were differentially affected by the three 18-carbon fatty acids. The cellular mRNA expressions of <i>ACSL3</i> and <i>FABP3</i> were increased by stearate, but were decreased by oleate or linoleate. However, the genes involved in fatty acid de novo synthesis (<i>ACACA</i> and <i>FASN</i>) and the regulation of transcription (<i>SREBP1</i>) were decreased by incubation with increasing concentrations of 18-carbon fatty acids. In conclusion, our findings provided evidence that 18-carbon fatty acids (stearate, oleate and linoleate) significantly increased cytosolic TAG accumulation in a dose-dependent manner, probably by promoting lipogenic genes and proteins that regulate the channeling of fatty acids towards milk TAG synthesis in pMECs.