Journal of Henan Agricultural Sciences ›› 2020, Vol. 49 ›› Issue (4): 160-166.DOI: 10.15933/j.cnki.1004-3268.2020.04.023
• Animal Husbandry and Veterinary Medicine • Previous Articles Next Articles
LIU Lili,YANG Bingyou
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刘莉莉,杨炳友
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Abstract: To investigate the mechanism of Rhaponticum uniflorum(L.)DC.regulating casein synthesis in mammary glands of mammary animal, in this study, the effects of ethanol extract from Rhaponticum uniflorum(L.)DC.(25,50,100,200,400,600,800 μg/mL)on the viability and proliferation ability of mammary epithelial cells in dairy goat were detected by using a cell activity analyzer.The concentration of ethanol extract not inhibiting cell growth and proliferation was selected to study the effect of ethanol extract on the mRNA expression of casein synthesis related genes of mammary epithelial cells in dairy goat.The mRNA expression levels of CSN1S1,CSN1S2,CSN2,CSN3,STAT5,JAK2,mTOR,4EBP1,S6K1 in mammary epithelial cells were detected by qRT-PCR. The results displayed that higher doses(200—800 μg/mL) of ethanol extract from Rhaponticum uniflorum(L.)DC. significantly inhibited the viability and proliferation ability of mammary epithelial cells in dairy goat (P<0.05).Thus,the epithelial cells were treated with 25,50,100 μg/ mL ethanol extract in subsequent experiments.The ethanol extract obviously up regulated the mRNA expression of CSN1S1,CSN1S2,CSN2,CSN3 genes and the genes related to casein synthesis such as STAT5,JAK2,mTOR,S6K1 (P<0.05). But the ethanol extract significantly down regulated the mRNA expression of 4EBP1 gene (P<0.05).This study shows that the ethanol extract from Rhaponticum uniflorum(L.) DC.can promote the synthesis of casein by regulating the expression of genes related to casein synthesis in mammary epithelial cells of dairy goats.
Key words: Rhaponticum uniflorum(L.)DC., Dairy goat, Mammary epithelial cells, αs1-casein, αs2-casein, β-casein, κ-casein, Gene expression
摘要: 为探讨漏芦对泌乳动物乳腺酪蛋白合成的调节机制,利用细胞活力分析仪检测漏芦乙醇提取物(25、50、100、200、400、600、800 μg/mL)对奶山羊乳腺上皮细胞活力和增殖能力的影响,并选择不抑制细胞生长增殖的乙醇提取物剂量,研究其对奶山羊乳腺上皮细胞酪蛋白合成相关基因mRNA表达的影响;采用实时荧光定量PCR(qRT-PCR)技术检测乳腺上皮细胞αs1-酪蛋白(CSN1S1)、αs2-酪蛋白(CSN1S2)、β-酪蛋白(CSN2)、κ-酪蛋白(CSN3)、信号转导与转录激活因子5(STAT5)、蛋白质酪氨酸激酶2(SJAK2)、哺乳动物雷帕霉素蛋白(mTOR)、真核细胞翻译启动因子4E结合蛋白1(4EBP1)及核糖体蛋白S6激酶1(S6K1)基因的mRNA表达水平。结果表明,较高剂量(200~800 μg/mL)的漏芦乙醇提取物明显抑制了奶山羊乳腺上皮细胞的活力和增殖能力(P<0.05),故后续试验添加25、50、100 μg/mL的漏芦乙醇提取物处理乳腺上皮细胞;漏芦乙醇提取物可明显上调细胞酪蛋白基因CSN1S1、CSN1S2、CSN2、CSN3及酪蛋白合成相关基因STAT5、JAK2、mTOR、S6K1的mRNA表达水平(P<0.05),但显著下调了4EBP1基因的mRNA表达水平(P<0.05)。由此可见,漏芦乙醇提取物能通过调节奶山羊乳腺上皮细胞酪蛋白合成相关基因的表达,进而促进乳腺细胞酪蛋白的合成。
关键词: 漏芦, 奶山羊, 乳腺上皮细胞, αs1-酪蛋白, αs2-酪蛋白, β-酪蛋白, κ-酪蛋白, 基因表达
CLC Number:
S823
LIU Lili, YANG Bingyou. Effect of Rhaponticum uniflorum(L.)DC.on the Expression of Genes Related to Casein Synthesis in Mammary Epithelial Cells of Dairy Goat[J]. Journal of Henan Agricultural Sciences, 2020, 49(4): 160-166.
刘莉莉, 杨炳友. 漏芦对奶山羊乳腺上皮细胞酪蛋白合成相关基因表达的影响[J]. 河南农业科学, 2020, 49(4): 160-166.
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URL: https://www.hnnykx.org.cn/EN/10.15933/j.cnki.1004-3268.2020.04.023
https://www.hnnykx.org.cn/EN/Y2020/V49/I4/160