Camellia (Camellia oleifera Abel.) seed oil is a commonly used edible oil of China. In ancient Chinese literature, it is mentioned to be helpful for postpartum repair and lactation in women. Research on camellia seed oil (CO) as a feed additive for dairy cattle is less. We investigated the effect of CO on the expression of milk fat and protein syntheses-related genes in differentiated bovine mammary epithelial cells (MAC-T) using soybean oil (SO) as the control. The results showed that CO increased the expression of genes related to de novo synthesis of fatty acids including sterol regulatory element-binding protein 1 (SREBP1), acetyl-CoA carboxylase 1 (ACC), fatty acid synthase (FASN), lipoprotein lipase (LPL), and stearoyl-CoA desaturase (SCD) (p < .05). Among the milk protein genes analyzed, CO increased β-casein mRNA expression (p < .05) and decreased αS1-casein mRNA expression (p < .05) in MAC-T cells. CO upregulated the pathways related to milk protein synthesis with increased mRNA levels of phosphoinositide 3-kinase (PI3K), RAC-alpha serine/threonine-protein kinase (AKT1), and mammalian target of rapamycin (mTOR) (p < .05) in MAC-T cells. Ribosomal protein S6 kinase beta-1 (S6K1) gene was upregulated, and eukaryotic initiation factor 4E (eIF4E) gene (p < .05) was downregulated with CO treatment.The mRNA expression levels of janus kinase 2 (JAK2), activator of transcription 5-β (STAT5-β), and E74-like factor 5 (ELF5) were elevated in MAC-T cells treated with CO (p < .05). Meanwhile, the protein expression levels of S6K1, STAT5-β, phosphorylated mTOR (p-mTOR), p-S6K1, and p-STAT5-β increased in MAC-T cells treated with CO (p < .05). In summary, CO promoted β-casein synthesis by regulating PI3K-mTOR-S6K1 and JAK2-STAT5 signaling pathways and influenced fatty acid synthesis by regulating SREBP1-related genes in MAC-T cells. We need to further confirm the function of CO using in vivo models.

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Objective: Caseins and fatty acids of milk are synthesized and secreted by the epithelial cells of the mammary gland. All-trans retinoic acid (ATRA), an active metabolite of vitamin A, has been shown to promote mammary development. This study was conducted to determine the effect of ATRA on casein synthesis and fatty acid composition in MAC-T cells.Methods: MAC-T cells were allowed to differentiate for 4 d, treated with ATRA (0, 1.0, 1.5, and 2.0 μM), and incubated for 3 d. We analyzed the fatty acid composition, the mRNA expression of casein and fatty acid synthesis-related genes, and the phosphorylation of casein synthesis-related proteins of MAC-T cells by gas chromatography, quantitative polymerase chain reaction, and western blotting, respectively.Results: In MAC-T cells, ATRA increased the mRNA levels of αS1-casein and β-casein, janus kinase 2 (JAK2) and E74-like factor 5 of the signal transducer and activator of transcription 5 β (STAT5-β) pathway, ribosomal protein S6 kinase beta-1 (S6K1) and eukaryotic translation initiation factor 4E binding protein 1 of the mammalian target of rapamycin (mTOR) pathway, inhibited the mRNA expression of phosphoinositide 3-kinase and eukaryotic initiation factor 4E of the mTOR pathway, and promoted the phosphorylation of STAT5-β and S6K1 proteins. Additionally, ATRA increased the de novo synthesis of fatty acids, reduced the content of long-chain fatty acids, the ratio of monounsaturated fatty acids to saturated fatty acids (SFA), the ratio of polyunsaturated fatty acids (PUFA) to SFA, and the ratio of ω-6 to ω-3 PUFA. The mRNA levels of acetyl-CoA carboxylase 1, fatty acid synthase, lipoprotein lipase, stearoyl-CoA desaturase, peroxisome proliferator-activated receptor gamma, and sterol regulatory element-binding protein 1 (SREBP1) were enhanced by ATRA.Conclusion: ATRA promotes the synthesis of casein by regulating JAK2/STAT5 pathway and downstream mTOR signaling pathway, and it improves the fatty acid composition of MAC-T cells by regulating SREBP1-related genes.