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Rapid communication: lipid metabolic gene expression and triacylglycerol accumulation in goat mammary epithelial cells are decreased by inhibition of SREBP-1.

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.

SUBMITTER: Xu H

PROVIDER: S-EPMC6095349 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature

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Rapid communication: lipid metabolic gene expression and triacylglycerol accumulation in goat mammary epithelial cells are decreased by inhibition of SREBP-1.

Xu Huifen H Luo Jun J Tian Huibin H Li Jun J Zhang Xueying X Chen Zhi Z Li Ming M Loor Juan J JJ

Journal of animal science 20180601 6

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 epithel ...[more]

PMID: 29846631

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Project description:Intramuscular fat (IMF) is closely related to the tenderness, juiciness, and flavor of beef, and is an important indicator for beef quality assessment internationally. The main components of skeletal intramuscular fat (IMF) are phospholipids and triacylglycerols (TAG), and the final step of TAG biosynthesis is catalyzed by diacylglycerol acyltransferase 2 (DGAT2). To explore the effect of DGAT2 on the differentiation of bovine muscle satellite cells (BSCs) and its role in the signaling pathway related to lipid metabolism, the adenovirus overexpression and interference vector of the DGAT2 gene was constructed in this study, and the overexpression adenovirus Ad-DGAT2 and interfering adenovirus sh-DGAT2 were used to infect BSCs. Overexpression of DGAT2 resulted in a significant increase in the contents of TAG and ADP, and the mRNA and protein expression levels of PPARγ, C/EBPα, and SREBF1 (p < 0.05). Interfering with the expression of DGAT2 reduced the intracellular TAG content and lipid droplet accumulation. Furthermore, the mRNA and protein expression levels of PPARγ, C/EBPα, and SREBF1 (p < 0.05) were significantly downregulated. Transcriptome sequencing showed that a total of 598 differentially expressed genes (DEGs) were screened in BSCs infected with Ad-DGAT2, and these DEGs included 292 upregulated genes and 306 downregulated genes. A total of 49 DEGs were screened in BSCs infected with sh-DGAT2, and these DEGs included 25 upregulated and 24 downregulated genes. KEGG enrichment analysis showed that the DEGs, after overexpression of DGAT2, were mainly enriched in the PPAR signaling pathway, and the fat digestion and absorption, glycerophospholipid metabolism, fatty acid biosynthesis, and AMPK signaling pathways. The DEGs obtained after interfering with DGAT2 were mainly enriched in the metabolic pathways, such as the PPAR signaling pathway and PI3K/AKT signaling pathway. In summary, our study demonstrated that the lipid droplet formation, TAG accumulation, and adipogenic gene expression in BSCs overexpressing DGAT2 were higher than those in the control cells. These results highlight the important role of DGAT2 in regulating BSCs during adipogenic transdifferentiation and underscore the complexity of intramuscular adipogenesis.

Dataset Information

Rapid communication: lipid metabolic gene expression and triacylglycerol accumulation in goat mammary epithelial cells are decreased by inhibition of SREBP-1.

Publications

Rapid communication: lipid metabolic gene expression and triacylglycerol accumulation in goat mammary epithelial cells are decreased by inhibition of SREBP-1.

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