Effects of Arginine on Casein Synthesis in Bovine Mammary Epithelial Cells

CAO Yue; LI Dabiao; SUN Mei; MU Xiaojia; HAO Yihong; YANG Jing

doi:10.3969/j.issn.1006-267x.2022.08.057

Chinese Journal of Animal Nutrition >

2022 , Vol. 34 >Issue 8: 5374 - 5383

DOI: https://doi.org/10.3969/j.issn.1006-267x.2022.08.057

RESEARCH PAPER

Effects of Arginine on Casein Synthesis in Bovine Mammary Epithelial Cells

CAO Yue ,
LI Dabiao ,
SUN Mei ,
MU Xiaojia ,
HAO Yihong ,
YANG Jing

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Key Laboratory of Animal Nutrition and Feed Science at Universities of Inner Mongolia Autonomous Region, College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China

Received date: 2022-01-15

Online published: 2022-08-11

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Abstract

In this paper, bovine mammary epithelial cells (BMECs) were used as a model to study the effects of adding different concentrations of arginine on the synthesis of casein in BMECs. A single-factor completely randomized trial design was used. 0.70 mmol/L arginine (concentration of arginine in DMEM medium) was used as the control group, and the concentrations of arginine in experimental groups were 1.40, 2.80, 5.60 and 11.20 mmol/L, respectively, with 6 replicates in each group. MTT method was used to detect the proliferation rate of BMECs; enzyme linked immunosorbent assay (ELISA) kits were used to detect the activities of key enzymes of arginine metabolism in BMECs; real-time quantitative PCR (RT-qPCR) technology was used to detect the genes expression levels of arginine metabolic key enzyme, casein and mammalian target of rapamycin (mTOR) signaling pathway in BMECs; Western Blot technology was used to detect the expression levels of casein and the protein phosphorylation levels of mTOR signaling pathway. The results showed as follows: 1) when the concentration of arginine was 2.80 mmol/L, the proliferation rate of BMECs was significantly higher than that in control group and other arginine groups (P<0.05). When arginine concentration was 2.80 and 5.60 mmol/L, ornithine decarboxylase activity in BMECs was significantly higher than that in control group and other arginine groups (P<0.05). 2) Gene relative expression levels of αs1-casein, κ-casein, protein kinase B, mammalian target of rapamycin, ribosomal p70s6 kinase and eukaryotic translation initiation factor 4E-binding protein 1 in BMECs in 2.80 mmol/L arginine group were significantly higher than those in the control group (P＜0.05). 3) The αs1-casein level and the phosphorylation level of S6K protein in BMECs in 1.40 and 2.80 mmol/L arginine groups were significantly higher than those in the control group and other arginine groups (P＜0.05). 4) The protein phosphorylation levels of mTOR signaling pathway in BMECs in 2.80 and 5.60 mmol/L arginine groups was significantly higher than that in control group and other arginine groups (P＜0.05). Based on the above results, when the concentration of arginine is 2.80 mmol/L, the promotion effect on the synthesis of casein in BMECs is the best.

Key words： arginine; bovine mammary epithelial cells; casein

Cite this article

CAO Yue , LI Dabiao , SUN Mei , MU Xiaojia , HAO Yihong , YANG Jing . Effects of Arginine on Casein Synthesis in Bovine Mammary Epithelial Cells[J]. Chinese Journal of Animal Nutrition, 2022 , 34(8) : 5374 -5383 . DOI: 10.3969/j.issn.1006-267x.2022.08.057

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