Effects of Histidine on Expressions of &beta;-casein, and Janus Kinases 2-Signal Transducer and Activator of Transcription 5/Mammalian Target Rapamycin Signaling Pathways Related Phospho-Proteins of in Vitro Cultured Bovine Mammary Epithelial Cells

WANG Shanshan, GAO Haina, ZHAO Shengguo, ZHENG Nan, ZHANG Yangdong, WANG Jiaqi, YAN Sumei

doi:10.3969/j.issn.1006-267x.2016.03.034

2016 , Vol. 28 >Issue 3: 916 - 925

DOI: https://doi.org/10.3969/j.issn.1006-267x.2016.03.034

Short Communications

Effects of Histidine on Expressions of β-casein, and Janus Kinases 2-Signal Transducer and Activator of Transcription 5/Mammalian Target Rapamycin Signaling Pathways Related Phospho-Proteins of in Vitro Cultured Bovine Mammary Epithelial Cells

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1. College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China;
2. Ministry of Agriculture-Milk and Dairy Product Inspection Center, Beijing 100193, China;
3. State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received date: 2015-10-13

Online published: 2016-03-14

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Abstract

The aim of this study was to investigate the effects of supplementation of different concentrations of histidine on expressions of β-casein and Janus kinases 2 (JAK2)-signal transducer and activator of transcription 5 (STAT5)/mammalian target rapamycin (mTOR) signaling pathways related phospho-proteins of in vitro cultured bovine mammary epithelial cells. The primary bovine mammary epithelial cells were used in this study, the special culture medium without essential amino acid (EAA) was used. Supplementation levels of histidine were 0 (control), 0.15, 0.60, 1.20, 2.40, 4.80, 9.60 and 19.20 mmol/L, respectively. After cultured for 12 h, primary bovine mammary epithelial cells proliferation was dectected by thiazolyl blue (MTT) method, the expressions of β-casein and 8 signaling pathway related phospho-proteins were determined by Western-Blot. The results showed as follows: 1) compared with control group the bovine mammary epithelial cell proliferation was increased after the supplementation of 0.15 to 9.60 mmol/L of histidine. 2) With the increase histidine concentration, the expression of β-casein measured tended to be increased at first and then decrease, and all experimental groups were significantly higher than control group (P<0.01). 3) Compared with control group, the supplementation of histidine could significantly promote the expressions of signaling pathway related phospho-proteins (P<0.01); among experimental groups, the expressions of phospho-mTOR [P-mTOR (Ser²⁴⁸¹)] and phospho-eukaryotic translation elongation factor 2 [P-eEF2 (Thr⁵⁶)] were reduced along with the increasing histidine concentration, and the expression of phospho-ribosome protein subunit 6 kinase 1 [P-S6K1 (Thr³⁸⁹)] was increased with the increasing histidine concentration; the expression of phospho-JAK2 [P-JAK2(Tyr^1007/1008)], phospho-eukaryotic initiation factor 4E binding protein [P-4EBP1(Thr³⁷)] and phospho-eukaryotic initiation factor 2α [P-eIF2α (Ser⁵¹)] were the highest at the supplementation of 2.40 mmol/L histidine, and the expressions of phospho-STAT5 [P-STAT5(Tyr⁶⁹⁴)], phospho-mTOR raptor [P-raptor(Ser⁸⁶³)] and mTOR complex binding protein (GβL) were highest at the supplementation of 9.60 mmol/L histidine. These results demonstrate that histidine can promote the expression of β-casein via P-JAK2(Tyr^1007/1008) and P-STAT5(Tyr⁶⁹⁴) of JAK2-STAT5 signaling pathway. Histidine at optimal concentration (0.15 to 9.60 mmol/L) range can also promote the expression of β-casein via P-raptor (Ser⁸⁶³) of mTORC1, which act on down-stream targets P-4EBP1 (Thr³⁷), and finally regulation of milk protein synthesis.

Key words： histidine; cell proliferation; JAK2; mTOR; β-casein

Cite this article

WANG Shanshan, GAO Haina, ZHAO Shengguo, ZHENG Nan, ZHANG Yangdong, WANG Jiaqi, YAN Sumei . Effects of Histidine on Expressions of β-casein, and Janus Kinases 2-Signal Transducer and Activator of Transcription 5/Mammalian Target Rapamycin Signaling Pathways Related Phospho-Proteins of in Vitro Cultured Bovine Mammary Epithelial Cells[J]. Chinese Journal of Animal Nutrition, 2016 , 28(3) : 916 -925 . DOI: 10.3969/j.issn.1006-267x.2016.03.034

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