Heo Young Tae, Ha Woo Tae, Lee Ran, Lee Won-Young, Jeong Ha Yeon, Hwang Kyu Chan, Song Hyuk
Department of Stem Cell and Regenerative Biology, College of Animal Bioscience and Technology, Konkuk University, Seoul 143-701, Korea.
Division of Food Bioscience, RIBHS, College of Biomedical and Health Sciences, Konkuk University, Chung-ju 380-701, Korea.
Asian-Australas J Anim Sci. 2017 Jun;30(6):878-885. doi: 10.5713/ajas.16.0515. Epub 2016 Sep 19.
Glucose is an essential fuel in the energy metabolism and synthesis pathways of all mammalian cells. In lactating animals, glucose is the major precursor for lactose and is a substrate for the synthesis of milk proteins and fat in mammary secretory (alveolar) epithelial cells. However, clear utilization of glucose in mammary cells during lactogenesis is still unknown, due to the lack of analyzing models. Therefore, the objective of this study was to test the reliability of the mammary alveolar (MAC-T) cell as an study model for glucose metabolism and lactating system.
Undifferentiated MAC-T cells were cultured in three types of Dulbecco's modified Eagle's medium with varying levels of glucose (no-glucose: 0 g/L, low-glucose: 1 g/L, and high-glucose: 4.5 g/L) for 8 d, after which differentiation to casein secretion was induced. Cell proliferation and expression levels of apoptotic genes, Insulin like growth factor-1 (IGF1) receptor, oxytocin receptor, αS1, αS2, and β casein genes were analyzed at 1, 2, 4, and 8 d after differentiation.
The proliferation of MAC-T cells with high-glucose treatment was seen to be significantly higher. Expression of apoptotic genes was not affected in any group. However, expression levels of the mammary development related gene (IGF1 receptor) and lactation related gene (oxytocin receptor) were significantly higher in the low-glucose group. Expressions of αS1-casein, αS2-casein, and β-casein were also higher in the low-glucose treated group as compared to that in the no-glucose and high-glucose groups.
The results demonstrated that although a high-glucose environment increases cell proliferation in MAC-T cells, a low-glucose treatment to MAC-T cells induces higher expression of casein genes. Our results suggest that the MAC-T cells may be used as an model to analyze mammary cell development and lactation connected with precise biological effects.
葡萄糖是所有哺乳动物细胞能量代谢和合成途径中的重要燃料。在泌乳动物中,葡萄糖是乳糖的主要前体,也是乳腺分泌(腺泡)上皮细胞合成乳蛋白和脂肪的底物。然而,由于缺乏分析模型,泌乳过程中乳腺细胞对葡萄糖的具体利用情况仍不清楚。因此,本研究的目的是测试乳腺腺泡(MAC-T)细胞作为葡萄糖代谢和泌乳系统研究模型的可靠性。
将未分化的MAC-T细胞在三种不同葡萄糖水平的杜氏改良 Eagle 培养基中培养8天(无葡萄糖:0 g/L,低葡萄糖:1 g/L,高葡萄糖:4.5 g/L),之后诱导其分化为酪蛋白分泌细胞。在分化后的第1、2、4和8天分析细胞增殖以及凋亡基因、胰岛素样生长因子-1(IGF1)受体、催产素受体、αS1、αS2和β酪蛋白基因的表达水平。
高葡萄糖处理的MAC-T细胞增殖明显更高。各实验组的凋亡基因表达均未受影响。然而,低葡萄糖组中乳腺发育相关基因(IGF1受体)和泌乳相关基因(催产素受体)的表达水平显著更高。与无葡萄糖组和高葡萄糖组相比,低葡萄糖处理组中αS1-酪蛋白、αS2-酪蛋白和β-酪蛋白的表达也更高。
结果表明,虽然高葡萄糖环境会增加MAC-T细胞的增殖,但对MAC-T细胞进行低葡萄糖处理会诱导酪蛋白基因的更高表达。我们的结果表明,MAC-T细胞可用作分析与精确生物学效应相关的乳腺细胞发育和泌乳的模型。