張興夫 高民 杜瑞平 敖長金 張航
摘 要:該研究旨在驗(yàn)證氨基酸模式的不同是否會影響奶牛乳腺上皮細(xì)胞酪蛋白的合成,并對其機(jī)理進(jìn)行初步探討。試驗(yàn)采用完全隨機(jī)試驗(yàn)設(shè)計,氨基酸模式分別為:低蛋白日糧條件下血液氨基酸組成模式組(LPBP)、全乳蛋白氨基酸組成模式組(MP)、酪蛋白氨基酸組成模式組(CP)、80%酪蛋白+20%乳清蛋白氨基酸組成模式組(CLP),每個處理3個重復(fù),試驗(yàn)重復(fù)3遍(n=12)。用RT-qPCR檢測CSN1S1、CSN3、GLUT1、PRLR、STAT5和mTOR基因表達(dá)量,用Elisa法檢測αS-酪蛋白合成量,用CCK-8檢測細(xì)胞增殖活性。試驗(yàn)結(jié)果顯示:氨基酸模式的不同影響奶牛乳腺上皮細(xì)胞酪蛋白的合成,全乳蛋白模式可能是一種較為理想的氨基酸模式。氨基酸模式通過調(diào)節(jié)mTOR、STAT5、PRLR和GLUT1基因的表達(dá)影響酪蛋白的合成。此外,研究發(fā)現(xiàn)在體外培養(yǎng)條件下,乳腺上皮細(xì)胞增殖活性與乳蛋白基因表達(dá)的趨勢并不一致。
關(guān)鍵詞:乳腺上皮細(xì)胞 氨基酸模式 基因表達(dá) 酪蛋白
Abstract:The objective of this study was to determine whether different amino acid (AA) patterns could affect casein synthesis in bovine mammary epithelial cells. A completely random design was used. Four AA patterns reatments are as follows: blood AA pattern in low protein diet (LPBP); milk protein AA pattern (MP); 80% casein plus 20% lactoalbumin AA pattern (CLP); casein AA pattern (CP). Each treatment has three replicates, and all experiments were repeated three times. The expression of CSN1S1, CSN3, GLUT1, PRLR, STAT5, mTOR genes and the content of αS-casein were determined by RT-QPCR method and αS -casein ELISA Kit, respectively.Cell proliferation was determined by CCK-8 method.The results showed that: The milk protein AA pattern might increase major milk protein genes expression. Perhaps, an appropriate AA pattern can promote mTOR STAT5 PRLR GLUT1 genes expression, and result in increased milk protein synthesis. In addition, the level of geng expression trend was different from the proliferation in mammary gland epithelial cell.
Key Words:Mammary Epithelial Cells;Amino Acid Pattern;Gene Expression;Casein
閱讀全文鏈接(需實(shí)名注冊):http://www.nstrs.cn/xiangxiBG.aspx?id=49048&flag=1