谷氨酰胺及其二肽对过氧化氢诱导山羊瘤胃上皮细胞凋亡及凋亡相关基因表达的影响

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(P0.05),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(P0.05)。2)与对照组相比,4组晚期凋亡的凋亡率显著增加(P0.05),试验组早期凋亡的凋亡率均显著增加(P0.05)。3)与对照组相比,试验组Bcl-2/Bax均显著增加(P0.05);与1组相比,2组、3组和4组Bcl-2/Bax均显著增加(P0.05),且2组显著高于3组、4组(P0.05)。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。

Effects of Glutamine and Its Dipeptides on Apoptosis and Apoptosis Related Gene Expressions Induced by Hydrogen Peroxide in Ruminal Epithelial Cells of Goats

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 (H2O2). 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 H2O2, 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 H2O2, and groups 2, 3 and 4 were cultured with 800 μmol/L H2O2, 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 H2O2 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 H2O2 concentration, while compared with control group, experimental groups were all significantly increased ( P<0.05) . 2) Compared with control group, apoptosis rate of late ap-optosis 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 H2O2 in ruminal epithe-lium cells of goats.