本文選題：蛋氨酸三肽　切入點：奶牛乳腺上皮細胞　出處：《動物營養(yǎng)學報》2017年09期 　論文類型：期刊論文

【摘要】：本試驗旨在研究蛋氨酸三肽(Met-Met-Met)對奶牛乳腺上皮細胞(BMECs)酪蛋白和小肽轉運載體基因表達量的影響。采用酶消化法培養(yǎng)的第3代奶牛乳腺上皮細胞為模型,各處理在培養(yǎng)基中分別添加0(對照)、40、50、60、70和80μg/mL的蛋氨酸三肽,每個處理5個重復,每個重復1個培養(yǎng)孔,分別培養(yǎng)細胞24、48和72h,檢測奶牛乳腺上皮細胞的相對增殖率,整體試驗重復2次,確定最佳培養(yǎng)時間;各處理在培養(yǎng)基中分別添加0(對照)、40、50、60、70和80μg/mL的蛋氨酸三肽,每個處理3個重復,每個重復1個培養(yǎng)孔,以最佳培養(yǎng)時間培養(yǎng),用實時定量PCR法檢測酪蛋白基因的表達量,確定適宜蛋氨酸三肽濃度,整體試驗重復3次;以最佳培養(yǎng)時間和適宜蛋氨酸三肽濃度培養(yǎng)細胞,以未添加蛋氨酸三肽的培養(yǎng)基為對照,每個處理3個重復,每個重復1個培養(yǎng)孔,測定小肽轉運載體基因的表達量,整體試驗重復3次。結果表明:在培養(yǎng)基中添加蛋氨酸三肽培養(yǎng)奶牛乳腺上皮細胞24h時,相對增殖率最高;培養(yǎng)基中加入60μg/mL的蛋氨酸三肽培養(yǎng)細胞24h,αs1-酪蛋白和β-酪蛋白的基因表達量最高,同時發(fā)現(xiàn)奶牛乳腺上皮細胞中小肽轉運載體1和小肽轉運載體2基因表達量顯著高于對照處理(P0.05)。綜上所述,培養(yǎng)基中添加60μg/mL的蛋氨酸三肽能夠提高奶牛乳腺上皮細胞酪蛋白和肽轉運載體基因的表達量。
[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.

【作者單位】： 內(nèi)蒙古農(nóng)業(yè)大學動物科學學院;
【基金】：國家奶業(yè)“973計劃”項目(2011CB100803)
【分類號】：S823.5

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