蛋氨酸三肽对奶牛乳腺上皮细胞酪蛋白和小肽转运载体基因表达量的影响  

Effects of Methionine Tripeptide on Expression Levels of Casein and Small Peptide Transporters Genes in Bovine Mammary Epithelial Cells

作　　者：郭春利[1] 丹妮[1] 曹琪娜[1] 哈斯额尔顿[1] 敖长金[1] 

机构地区：[1]内蒙古农业大学动物科学学院,呼和浩特010018

出　　处：《动物营养学报》2017年第9期3159-3166,共8页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家奶业"973计划"项目(2011CB100803)

摘　　要：本试验旨在研究蛋氨酸三肽(Met-Met-Met)对奶牛乳腺上皮细胞(BMECs)酪蛋白和小肽转运载体基因表达量的影响。采用酶消化法培养的第3代奶牛乳腺上皮细胞为模型,各处理在培养基中分别添加0(对照)、40、50、60、70和80μg/mL的蛋氨酸三肽,每个处理5个重复,每个重复1个培养孔,分别培养细胞24、48和72h,检测奶牛乳腺上皮细胞的相对增殖率,整体试验重复2次,确定最佳培养时间;各处理在培养基中分别添加0(对照)、40、50、60、70和80μg/mL的蛋氨酸三肽,每个处理3个重复,每个重复1个培养孔,以最佳培养时间培养,用实时定量PCR法检测酪蛋白基因的表达量,确定适宜蛋氨酸三肽浓度,整体试验重复3次;以最佳培养时间和适宜蛋氨酸三肽浓度培养细胞,以未添加蛋氨酸三肽的培养基为对照,每个处理3个重复,每个重复1个培养孔,测定小肽转运载体基因的表达量,整体试验重复3次。结果表明:在培养基中添加蛋氨酸三肽培养奶牛乳腺上皮细胞24h时,相对增殖率最高;培养基中加入60μg/mL的蛋氨酸三肽培养细胞24h,αs1-酪蛋白和β-酪蛋白的基因表达量最高,同时发现奶牛乳腺上皮细胞中小肽转运载体1和小肽转运载体2基因表达量显著高于对照处理(P<0.05)。综上所述,培养基中添加60μg/mL的蛋氨酸三肽能够提高奶牛乳腺上皮细胞酪蛋白和肽转运载体基因的表达量。This experiment aimed to study the effects of methionine tripeptide （Met-Met-Met） on expression levels casein and small peptide transporters genes in bovine mammary epithelial cells （BMECs）. Using BMECs isolated with collagenase digestion method as the model, different concentrations[0 （control）, 40, 50, 60, 70 and 80 μg/mL] of Met-Met-Met were added in culture medium in different treatments and cultured for 24, 48, and 72 h, respectively, and each treatment had 5 replicates with 1 culture pore per replicate. The experiment was repeated twice. Relative growth rate was measured to determine the optimal culture time. Different concentrations[0 （control）, 40, 50, 60, 70 and 80 μg/mL] of Met-Met-Met were added in culture medium in different treatments and cultured for the optimal time, and each treatment had 3 replicates with 1 culture pore per replicate. The experiment was repeated for three times. Expression levels of casein genes were determined by real-time PCR to determine the optimal Met-Met-Met concentration. Using the optimal culture time and Met-Met-Met concentration to culture cells, and culture medium without Met-Met-Met was used for control. Each treatment had 3 replicates with 1 culture pore per replicate. The experiment was repeated three times. Expression levels of peptide transporter genes were examined. The results showed as follows：relative growth rate was the highest when cells were treated with Met-Met-Met for 24 h; with the suitable culturing time of 24 h, when adding 60 μg/mL Met-Met-Met, expression levels of αs1-casein and β-casein gens were the highest, meanwhile expression levels of small peptide transporter 1 and 2 genes were significantly higher than those in control treatment （P〈0.05）. To sum up, adding 60 μg/mL of Met-Met-Met can increase expression levels of casein and peptide transporter genes in BMECs.

关 键 词：蛋氨酸三肽 奶牛乳腺上皮细胞 酪蛋白 小肽转运载体 

分 类 号：S823[农业科学—畜牧学]