D-Glucose and amino acid deficiency inhibits casein synthesis through JAK2/STAT5 and AMPK/mTOR signaling pathways in mammary epithelial cells of dairy cows

文献类型： 外文期刊

第一作者： Zhang, M. C.

作者： Zhang, M. C.;Zhao, S. G.;Wang, S. S.;Luo, C. C.;Gao, H. N.;Zheng, N.;Wang, J. Q.;Zhang, M. C.;Zhao, S. G.;Wang, S. S.;Luo, C. C.;Gao, H. N.;Zheng, N.;Wang, J. Q.;Zhang, M. C.;Zhao, S. G.;Wang, S. S.;Luo, C. C.;Gao, H. N.;Zheng, N.;Wang, J. Q.;Zhang, M. C.;Zhao, S. G.;Wang, S. S.;Luo, C. C.;Gao, H. N.;Zheng, N.;Wang, J. Q.

作者机构：

关键词： glucose;amino acid;signaling pathway;milk protein

期刊名称：JOURNAL OF DAIRY SCIENCE （ 影响因子：4.034； 五年影响因子：4.354 ）

ISSN： 0022-0302

年卷期： 2018 年 101 卷 2 期

页码：

收录情况： SCI

摘要： Amino acids and energy deficiency lead to lower milk protein content in dairy cows. However, the known mechanisms involved in this process do not adequately explain the variability of milk protein concentration in the mammary gland. We hypothesized that a deficiency in D-glucose (D-Glc) or AA would inhibit casein synthesis by regulating signaling pathways in mammary epithelial cells. Cow mammary epithelial cells (CMEC) were subjected to combinations of 1 of 3 concentrations of D-Glc (0, 2.50, or 17.5 mM) and 1 of 3 concentrations of AA (0, 1.03, or 7.20 mM). The effect of each mixture on cell cycle stage was assessed by flow cytometry. The expression levels of beta-casein and.-casein (encoded by CSN2 and CSN3) were measured by quantitative real-time PCR and Western blotting. Phosphorylation of Janus kinase 2 (Jak2), signal transducer and activator of transcription 5a (Stat5a), AMP-activated protein kinase (AMPK), mammalian target of rapamycin (mTOR), ribosomal protein S6 kinase 1 (S6K1), and eukaryotic factor 4E-binding protein 1 (4EBP1) were analyzed by Western blotting. The percentages of cells in the DNA postsynthetic (G2) and DNA synthesis (S) phases would decrease, with the level of D-Glc or AA declining individually, but no interaction was observed between the D-Glc and AA effects. The CSN2 and CSN3 mRNA and protein were downregulated when D-Glc or AA decreased individually from 17.5 to 2.50 mM or from 7.20 to 1.03 mM, but D-Glc deficiency had a greater effect according to the regression analysis. The phosphorylation ratio of Jak2 (Tyr(1007/1008)), Stat5a (Tyr(694)), mTOR (Ser(2448)), S6K1 (Thr(389)), and 4EBP1 (Thr(37)) was downregulated with the level of D-Glc or AA decline, whereas the phosphorylation ratio of AMPK (Thr(183/172)) was upregulated. And the change of D-Glc level had a more marked effect than AA in regulating the activity of these signaling protein above according to the regression analysis. Thus, D-Glc or AA deficiency likely reduced casein transcription via inhibition of the Jak2/Stat5 pathway, and reduced translation via suppression of the mTOR pathway by activation of AMPK, but D-Glc deficiency had a more marked effect. These indicated that deficiency of AA, and especially Glc, suppressed proliferation of CMEC and casein gene and protein expression, associated with inhibition of JAK2/STAT5 and AMPK/mTOR signaling pathways.

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