Effects of L-Theanine on Apoptosis of Subculture Ruminal Epithelial Cells Induced by Hydrogen Peroxide of Goats

JIE Hongdong, HAN Qipeng, TAN Zhiliang, ZHOU Chuanshe, KONG Zhiwei, CHEN Liang, REN Ao

doi:10.3969/j.issn.1006-267x.2017.05.017

Chinese Journal of Animal Nutrition >

2017 , Vol. 29 >Issue 5: 1582 - 1589

DOI: https://doi.org/10.3969/j.issn.1006-267x.2017.05.017

Molecular Nutrition

Effects of L-Theanine on Apoptosis of Subculture Ruminal Epithelial Cells Induced by Hydrogen Peroxide of Goats

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1. College of Horticulture & Landscape, Hunan Agricultural University, Changsha 410128, China;
2. Key Laboratory for Agro-Ecological Processes in Subtropical Region, National Engineering Laboratory for Pollution Control and Waste Utilization in Livestock and Poultry Production, Hunan Research Center of Livestock & Poultry Sciences, South-Central Experimental Station of Animal Nutrition and Feed Science of Ministry of Agriculture, Institute of Subtropical Agriculture, The Chinese Academy of Science, Changsha 410125, China;
3. Hunan Co-Innovation Center of Animal Production Safety, Changsha 410128, China

Received date: 2017-01-27

Online published: 2017-05-13

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Abstract

The aim of this study was to explore the effects of L-theanine on the apoptosis ratio and expression quantities of key genes in apoptosis pathway of subculture ruminal epithelial cells induced by hydrogen peroxide (H₂O₂) of goats according to establish the apoptosis model for subculture ruminal epithelium cells induced by H₂O₂. Subculture ruminal epithelium cells of 42-day-old Xiangdong black goats were selected, and were cultured in DMEM/F12 medium with 5% FBS, when the intensity reached 60% to 70%, the cells were randomly divided into 5 groups: control group, with no supplementation in medium; group Ⅰ, supplemented with 800 μmol/L H₂O₂ in medium; group Ⅱ, supplemented with 800 μmol/L+4 mmol/L L-theanine in medium, group Ⅲ, supplemented with 800 μmol/L+8 mmol/L L-theanine in medium, group Ⅳ, supplemented with 800 μmol/L+16 mmol/L L-theanine in medium. There were three repeats in each group. After 12 h treatment, flow cytometry (FCM) was used to detect the apoptosis ratio of the subculture ruminal epithelium cells, and real-time fluorescent quantitative PCR (RT-qPCR) was selected to detect the expression quantities of apoptosis pathway key genes of subculture ruminal epithelial cells, such as cysteinyl aspartate-specific proteinase-3 (Caspase-3), cysteinyl aspartate-specific proteinase-8 (Caspase-8), cysteinyl aspartate-specific proteinase-9 (Caspase-9), apoptotic protease activating facter-1 (Apaf-1) and Fas-associate with death domain protein (FADD). The results showed as follows: 1) compared with the group Ⅰ, the late apoptosis ratio of cells for L-theanine supplementation groups (groups Ⅱ, Ⅲ and Ⅳ) decreased significantly (P<0.05), and the late apoptosis ratio decreased with the increase of L-theanine supplemental concentration by using annexin V/propidium iodide (PI) unite staining. 2) The expression quantities of Caspase-3, Caspase-9 and Apaf-1 for L-theanine supplementation groups decreased significantly compared with the group Ⅰ using RT-qPCR test (P<0.05). It is concluded that L-theanine plays a protection role in apoptosis subculture ruminal epithelium cells induced by H₂O₂ of goats, which may provide technical support and theoretical reference for the future research on oxidative stress damage mechanism of the rumen in ruminants.

Key words： L-theanine; H₂O₂; goats; subculture ruminal epithelium cell; apoptosis

Cite this article

JIE Hongdong, HAN Qipeng, TAN Zhiliang, ZHOU Chuanshe, KONG Zhiwei, CHEN Liang, REN Ao . Effects of L-Theanine on Apoptosis of Subculture Ruminal Epithelial Cells Induced by Hydrogen Peroxide of Goats[J]. Chinese Journal of Animal Nutrition, 2017 , 29(5) : 1582 -1589 . DOI: 10.3969/j.issn.1006-267x.2017.05.017

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