Effect of linoleic acid supplementation on triglyceride content and gene expression in milk fat synthesis in two- and three-dimensional cultured bovine mammary epithelial cells
Effect of linoleic acid supplementation on triglyceride content and gene expression in milk fat synthesis in two- and three-dimensional cultured bovine mammary epithelial cells
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DOI：
10.1080/1828051x.2017.1412273
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发表时间：
2017-12
影响因子：
2.5
通讯作者：
Dabiao Li;Y. Xing;Honglei Li;Weiyun Wang;X. Hou;M. Gao
中科院分区：
文献类型：
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作者：
Dabiao Li;Y. Xing;Honglei Li;Weiyun Wang;X. Hou;M. Gao
摘要本研究探讨了0、40和80 μmol/L乳酸对二维和三维培养的牛乳腺上皮细胞（BMEC）甘油三酯（TG）含量及乳脂合成相关基因mRNA表达的影响。用TG测定试剂盒测定TG含量，用实时定量聚合酶链反应（RT-qPCR）测定基因表达。加入80 μmol/L LA可显著增加2D和3D培养的BMEC细胞内TG含量（p <0.05）。加入80和40 μmol/L LA后，二酰基甘油酰基转移酶2（DGAT 2）和过氧化物酶体增殖物激活受体-γ（PPARG）的mRNA表达分别增加（p < .05），而PPARG的表达在2D模型中下调（p < .05）。与对照组相比，添加LA显著降低了固醇调节元件结合转录因子1（SREBF 1）、乙酰辅酶A羧化酶（ACC）、脂肪酸合成酶（FATs）、硬脂酰辅酶A去饱和酶（SCD）和脂肪酸结合蛋白3（FABP 3）的表达（p <0.05）。总之，在3D BMEC培养模型中，添加LA的BMEC中TG含量和乳脂合成相关基因的表达显著高于2D培养模型（p <0.05），40和80 µmol/L的LA抑制脂肪酸的从头合成，促进DGAT 2的表达和TG的积累。40 μmol/L LA可上调3D模型中PPARG mRNA的表达，而下调2D模型中PPARG mRNA的表达。
Abstract In this study, the effect of 0, 40, and 80 μmol/L of LA on triglyceride (TG) content was evaluated, as well as the mRNA expression of genes involved in milk fat synthesis in bovine mammary epithelial cells (BMECs) which were cultured in two-dimensional (2D) and three-dimensional (3D) models. TG content was measured with a TG determination kit, and the expression of genes was measured with real-time quantitative polymerase chain reactions (RT-qPCR). Adding 80 μmol/L of LA significantly increased intracellular TG content in both 2D and 3D cultured BMECs (p < .05). The mRNA expression of diacylglycerol acyltransferase 2 (DGAT2) and peroxisome proliferator-activated receptor-γ (PPARG) increased with the addition of 80 and 40 μmol/L of LA, respectively (p < .05), whereas the expression of PPARG was downregulated in the 2D model (p < .05). Adding LA significantly reduced the expression of sterol regulatory element-binding transcription factor 1 (SREBF1), acetyl-CoA carboxylase (ACC), fatty acid synthase (FASN), stearoyl-CoA desaturase (SCD), and fatty acid-binding protein 3 (FABP3) compared to the control group (p < .05). Altogether, TG content in BMECs and expression of genes involved in milk fat synthesis were significantly higher with addition of LA under 3D BMECs culture model than those of 2D (p < .05), and 40 and 80 µmol/L of LA inhibited the de novo synthesis of fatty acids and promoted the expression of DGAT2 and accumulation of TG. The mRNA expression of PPARG in 3D model was increased by addition of 40 μmol/L LA, whereas it was downregulated in 2D model.