本文選題：蛋氨酸三肽　切入點(diǎn)：奶牛乳腺上皮細(xì)胞　出處：《動(dòng)物營養(yǎng)學(xué)報(bào)》2017年09期 　論文類型：期刊論文

【摘要】：本試驗(yàn)旨在研究蛋氨酸三肽(Met-Met-Met)對(duì)奶牛乳腺上皮細(xì)胞(BMECs)酪蛋白和小肽轉(zhuǎn)運(yùn)載體基因表達(dá)量的影響。采用酶消化法培養(yǎng)的第3代奶牛乳腺上皮細(xì)胞為模型,各處理在培養(yǎng)基中分別添加0(對(duì)照)、40、50、60、70和80μg/mL的蛋氨酸三肽,每個(gè)處理5個(gè)重復(fù),每個(gè)重復(fù)1個(gè)培養(yǎng)孔,分別培養(yǎng)細(xì)胞24、48和72h,檢測(cè)奶牛乳腺上皮細(xì)胞的相對(duì)增殖率,整體試驗(yàn)重復(fù)2次,確定最佳培養(yǎng)時(shí)間;各處理在培養(yǎng)基中分別添加0(對(duì)照)、40、50、60、70和80μg/mL的蛋氨酸三肽,每個(gè)處理3個(gè)重復(fù),每個(gè)重復(fù)1個(gè)培養(yǎng)孔,以最佳培養(yǎng)時(shí)間培養(yǎng),用實(shí)時(shí)定量PCR法檢測(cè)酪蛋白基因的表達(dá)量,確定適宜蛋氨酸三肽濃度,整體試驗(yàn)重復(fù)3次;以最佳培養(yǎng)時(shí)間和適宜蛋氨酸三肽濃度培養(yǎng)細(xì)胞,以未添加蛋氨酸三肽的培養(yǎng)基為對(duì)照,每個(gè)處理3個(gè)重復(fù),每個(gè)重復(fù)1個(gè)培養(yǎng)孔,測(cè)定小肽轉(zhuǎn)運(yùn)載體基因的表達(dá)量,整體試驗(yàn)重復(fù)3次。結(jié)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(xì)胞24h時(shí),相對(duì)增殖率最高;培養(yǎng)基中加入60μg/mL的蛋氨酸三肽培養(yǎng)細(xì)胞24h,αs1-酪蛋白和β-酪蛋白的基因表達(dá)量最高,同時(shí)發(fā)現(xiàn)奶牛乳腺上皮細(xì)胞中小肽轉(zhuǎn)運(yùn)載體1和小肽轉(zhuǎn)運(yùn)載體2基因表達(dá)量顯著高于對(duì)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(xì)胞酪蛋白和肽轉(zhuǎn)運(yùn)載體基因的表達(dá)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(xué)動(dòng)物科學(xué)學(xué)院;
【基金】：國家奶業(yè)“973計(jì)劃”項(xiàng)目(2011CB100803)
【分類號(hào)】：S823.5

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