作 者：
孔志伟;揭红东;陈亮;任傲;周传社;谭支良
关键词：
L-茶氨酸;过氧化氢;瘤胃上皮细胞;凋亡
摘 要：
本试验旨在研究L-茶氨酸对过氧化氢(H_2O_2)诱导山羊瘤胃上皮细胞凋亡的保护作用。采集42日龄的湘东黑山羊的瘤胃组织进行原代细胞分离、培养、纯化。对照组采用完全培养基,Ⅰ、Ⅱ、Ⅲ、Ⅳ组分别在完全培养基中添加800μmol/L H_2O_2、4 mmol/L L-茶氨酸+800μmol/L H_2O_2、8 mmol/L L-茶氨酸+800μmol/L H_2O_2和16 mmol/L L-茶氨酸+800μmol/L H_2O_2,每组3个重复。培养12 h后,采用噻唑蓝(MTT)法检测细胞存活率,采用流式细胞术检测细胞周期,采用实时定量PCR法和蛋白质印迹(Western blot)法检测Bax和Bcl-2基因及其蛋白的表达。结果表明:L-茶氨酸能显著提高H_2O_2损伤的瘤胃上皮细胞存活率(P<0.05),显著增加S期细胞比例(P<0.05),显著降低Bax基因和蛋白表达量(P<0.05),显著提高Bcl-2基因和蛋白表达量(P<0.05),显著提高Bcl-2和Bax基因和蛋白表达量的比值(P<0.05)。结果提示,L-茶氨酸能通过抑制细胞凋亡来有效地保护由H_2O_2导致的瘤胃上皮细胞损伤;体外条件下,培养液中添加的L-茶氨酸浓度高于8 mm/L时能有效发挥抗凋亡作用。对瘤胃上皮细胞损伤有一定的治疗和保护作用。
译 名：
Protection of L-Theanine on Apoptosis of Rumen Epithelial Cells Induced by Hydrogen Peroxide H_2O_2 in Goats
作 者：
KONG Zhiwei;JIE Hongdong;CHEN Liang;REN Ao;ZHOU Chuanshe;TAN Zhiliang;University of the Chinese Academy of Sciences;Key Laboratory for Agro-Ecological Processes in Subtropical Region,National Engineering Laboratory for Pollution Control and Waste Utilization in Livestock and Poultry Production,South-Central Experimental Station of Animal Nutrition and Feed Science in Ministry of Agriculture ,Hunan Research Center of Livestock &Poultry Sciences ,Institute of Subtropical Agriculture ,The Chinese Academy of Sciences;College of Animal Science and Technology,Hunan Agricultural University;Hunan Co-Innovation Center of Safety Animal Production;
单 位：
KONG Zhiwei%JIE Hongdong%CHEN Liang%REN Ao%ZHOU Chuanshe%TAN Zhiliang%University of the Chinese Academy of Sciences%Key Laboratory for Agro-Ecological Processes in Subtropical Region,National Engineering Laboratory for Pollution Control and Waste Utilization in Livestock and Poultry Production,South-Central Experimental Station of Animal Nutrition and Feed Science in Ministry of Agriculture ,Hunan Research Center of Livestock &Poultry Sciences ,Institute of Subtropical Agriculture ,The Chinese Academy of Sciences%College of Animal Science and Technology,Hunan Agricultural University%Hunan Co-Innovation Center of Safety Animal Production
关键词：
L-theanine;;hydrogen peroxide;;rumen epithelial cells;;apoptosis
摘 要：
The object of this experiment was to explore the protection of L-theanine on apoptosis of rumen epithelial cells induced by hydrogen peroxide( H_2O_2) in goats. Ruminal epithelial tissue from Xiangdong black goats aged 42 days were selected for isolation,cultivation and purification of primary cells. The cells were cultured in a complete culture medium in control group,and those in groups Ⅰ,Ⅱ,Ⅲ and Ⅳ were cultured in the complete culture medium supplemented with 800 μmol/L H_2O_2,4 mmol/L L-theanine + 800 μmol/L H_2O_2,8 mmol/L L-theanine+800 μmol/L H_2O_2,and 16 mmol/L L-theanine + 800 μmol/L H_2O_2,respectively. Each group had 3 replicates. After 12 h culture,thiazolyl blue( MTT) method was used to detect the cells viability,flowcytometry( FCM) was used to detect the cell cycle,real-time quantitative PCR and Western blot method were used to detect expressions of Bcl-2 and Bax genes and their proteins. The results showed that L-theanine significantly attenuated the cell viability loss( P < 0.05),significantly increased the cell ratio in S phrase( P < 0.05),significantly decreased expression levels of Bax gene and protein( P < 0.05),significantly increased expression levels of Bcl-2 gene and protein( P < 0. 05),and significantly increased the ratios of Bax gene to Bcl-2 gene and Bax protein to Bcl-2 protein in rumen epithelial cells injured by H_2O_2( P<0.05).The results indicate that L-theanine can prevent rumen epithelial cells from H_2O_2-induced cell apoptosis. Under in vitro conditions,the supplementation of L-theanine more than 8 mm/L can effectively help to protect the rumen epithelial cells from apoptosis.
