Abstract
In mammals, sterol regulatory element binding protein-1 (SREBP-1) is the master regulator of fatty acid and triacylglycerol synthesis. Recent gene silencing studies in mammary cells indicate that SREBP-1 has a central role in milk fat synthesis. However, SREBP-1 knockdown studies in goat mammary cells have not been performed; hence, its direct role in controlling mRNA expression of lipid metabolism genes and triacylglycerol synthesis remains unknown. Inhibition of SREBP-1 in goat mammary epithelial cells (GMEC) by small interference RNA (siRNA) markedly reduced the content of cellular triacylglycerol (~50% decrease, P < 0.05) and was partly related to downregulation of AGPAT6, LPIN1, and DGAT2 (-23%, -28% and -19%, respectively. P < 0.05), which are key enzymes involved in triacylglycerol synthesis, cellular triacylglycerol content and lipid droplet accumulation all decreased by SREBP-1 inhibition. The expression of lipid droplet formation and secretion genes was not altered in response to treatment. Although the lack of effect on expression of ACACA and FASN (rate-limiting enzymes for de novo fatty acid synthesis) with SREBP-1 knockdown was unexpected (P > 0.05), the downregulation of genes related to synthesis of acetyl-CoA and acetate activation (ACLY, ACSS2, and IDH1, P < 0.05) suggests that lipogenesis was inhibited. SREBP-1 knockdown also resulted in decreased expression of genes associated with fatty acid desaturation and elongation (SCD1 and ELOVL6, P < 0.05), long-chain fatty acid (LCFA) activation and transport (ACSL1, FABP3, and SLC27A6, P < 0.05). The results underscored the essential role of SREBP-1 not only in fatty acid synthesis but also in desaturation, elongation, and esterification in GMEC. Clearly, the lack of effect on ACACA and FASN, both of which are considered the key lipogenic enzymes, implies that there may be different regulatory mechanisms in goat compared with bovine mammary cells.