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

发布时间：2018-03-19 13:01

本文选题：蛋氨酸三肽　切入点：奶牛乳腺上皮细胞　出处：《动物营养学报》2017年09期 　论文类型：期刊论文

【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(P0.05)。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。
[Abstract]:This experiment was conducted to study the methionine three peptide (Met-Met-Met) on bovine mammary epithelial cells (BMECs) and casein peptide transporter gene expression. The third generation of bovine mammary epithelial cells cultured by enzyme digestion method for the model, each treatment in medium were added 0 training (control), 40,50,60,70 and 80 g/mL three methionine peptides, with 5 replicates of each repeat of 1 wells, respectively 24,48 and 72h cells, detect the relative proliferation of bovine mammary epithelial cells, the whole experiment was repeated 2 times, to determine the optimum culture time; each treatment in the medium were added 0 training (control) and 80 g/mL 40,50,60,70. Three methionine peptides, with 3 replicates of each repeat of 1 wells, with the best training time culture, the expression of real-time quantitative PCR assay of casein gene, determine the appropriate methionine concentration of three peptide, the whole experiment was repeated 3 times with; The optimal incubation time and suitable methionine concentration of three peptide cell culture medium without methionine peptide three as control, with 3 replicates of each repeat 1 wells, expression quantity of small peptide transporter gene, the whole experiment was repeated 3 times. The results showed that: in cultured mammary epithelial cells of cow 24h methionine three peptide medium, the relative growth rate is highest; 60 g/mL added to the culture medium of methionine three peptide and cultured 24h cells, alpha s1- casein and beta casein gene expression was the highest, while that of bovine mammary epithelial cells of small peptide transporter 1 and peptide transporter 2 gene expression was significantly higher than that of the control treatment (P0.05). To sum up, 60 g/mL added to the culture medium can improve the expression of methionine three peptides of bovine mammary epithelial cells and casein peptide transporter gene.

【作者单位】：内蒙古农业大学动物科学学院;
【基金】：国家奶业“973计划”项目(2011CB100803)
【分类号】：S823.5

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