作 者：
韩奇鹏;掲红东;罗玲;王凯军;周传社;张佩华;孔志伟;汤少勋
关键词：
湘东黑山羊;瘤胃上皮细胞;凋亡;凋亡基因
摘 要：
本试验通过建立过氧化氢(H2O2)诱导山羊瘤胃上皮传代细胞凋亡模型,研究谷氨酰胺(Gln)、甘氨酰谷氨酰胺(Gly-Gln)和丙氨酰谷氨酰胺(Ala-Gln)对凋亡细胞的凋亡率及Bcl-2、Bax基因表达量的影响。选用60日龄湘东黑山羊的瘤胃上皮传代细胞,采用不同浓度[0(对照组)、100、400、800μmol/L]的H2O2培养细胞,应用流式细胞术检测细胞凋亡情况。传代瘤胃上皮细胞分为5组,对照组和1组分别添加0、800μmol/L H2O2,2组、3组、4组均添加800μmol/L H2O2,同时分别添加17.28 mmol/L Gly-Gln(2组)、16.0 mmol/L Gln(3组)、16.0 mmol/L AlaGln(4组),应用流式细胞术检测细胞凋亡情况,同时采用实时荧光定量PCR(FQ-PCR)法检测细胞Bcl-2、Bax基因表达量。结果显示:1)与对照组相比,当H2O2浓度增加到800μmol/L时,早期凋亡的凋亡率显著增加(P<0.05),而晚期凋亡的凋亡率随着H2O2浓度的增加呈现增加后减少的趋势,但相对于对照组,都呈显著增加(P<0.05)。2)与对照组相比,4组晚期凋亡的凋亡率显著增加(P<0.05),试验组早期凋亡的凋亡率均显著增加(P<0.05)。3)与对照组相比,试验组Bcl-2/Bax均显著增加(P<0.05);与1组相比,2组、3组和4组Bcl-2/Bax均显著增加(P<0.05),且2组显著高于3组、4组(P<0.05)。综合得出,Gly-Gln对H2O2引起山羊瘤胃上皮细胞早期凋亡具有一定的保护作用。
译 名：
Effects of Glutamine and Its Dipeptides on Apoptosis and Apoptosis Related Gene Expressions Induced by Hydrogen Peroxide in Ruminal Epithelial Cells of Goats
作 者：
HAN Qipeng;JIE Hongdong;LUO Ling;WANG Kaijun;ZHOU Chuanshe;ZHANG Peihua;KONG Zhiwei;TANG Shaoxun;Key Laboratory for Agro-Ecological Processes in Subtropical Region,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 ,Chinese Academy of Sciences;Hunan Provincial Key Laboratory for G enetic Improvement of Domestic Animal,College of Animal Science and Technology,Hunan Agricultural University;
关键词：
Xiangdong black goat;;ruminal epithelium cell;;apoptosis;;apoptosis gene
摘 要：
This study was conducted to investigate the effects of glutamine(Gln),glycyl-glutamine(Gly-Gln)and alanyl-glutamine(Ala-Gln) on apoptosis rate and gene expressions of Bcl-2 and Bax of apoptosis cells according to establish apoptosis models for ruminal epithelial cells of goats induced by hydrogen peroxide(H_2O_2).Subculture ruminal epithelium cells of 60 day-old Xiangdong black goats were selected and cultured with different concentrations [0(control group),100,400 and 800 μmol/L] of H_2O_2,and flow cytometry(FCM) technique was used to detected cell apoptosis.Subculture ruminal epithelium cells were divided into 5 groups,control group and group 1 were cultured with 0 and 800 μmol/L H_2O_2,and groups 2,3 and 4 were cultured with 800 μmol/L H_2O_2,meanwhile with 17.28 mmol/L Gly-Gln(group 2),16.0 mmol/L Gln(group 3) and 16.0 mmol/L Ala-Gln(group 4).FCM technique was used to detected cell apoptosis,and gene expressions of Bcl-2 and Bax were detected by real-time fluorescent quantitative PCR.The results showed as follows: 1) compared with control group,when the concentration of H_2O_2 reached 800 μmol/L,apoptosis rate of early apoptosis significantly increased(P < 0.05); apoptosis rate of late apoptosis firstly increased and then decreased with the increasing of H_2O_2 concentration,while compared with control group,experimental groups were all significantly increased(P< 0.05).2) Compared with control group,apoptosis rate of late apoptosis in group 4 significant increased(P <0.05),and apoptosis rate of early apoptosis in experimental groups were all significantly higher than that in control group(P < 0.05).3) Compared with control group,Bcl-2/Bax in experimental groups was significantly increased(P < 0.05); compared with group 1,Bcl-2/Bax in groups 2,3 and 4 was significantly increased(P <0.05),and group 2 was significantly higher than groups 3 and 4(P < 0.05).In conclusion,Gly-Gln plays protection role in early apoptosis induced by H_2O_2 in ruminal epithelium cells of goats.
