不同浓度亚油酸对二维和三维培养模式下奶牛乳腺上皮细胞形态及乳蛋白合成相关基因表达的影响  

Effects of Different Concentrations of Linoleic Acid on Expression of Milk Protein Synthesis Related Gene in Two-Dimension and Three-Dimension Cultured Bovine Mammary Epithelial Cells

作　　者：邢媛媛[1] 李大彪[1] 李子南 金亚亚 XING Yuanyuan;LI Dabiao;LI Zinan;JIN Yaya(College of Animal Science,Inner Mongolia Agricultural University,Hohhot 010018,China)

机构地区：[1]内蒙古农业大学动物科学学院

出　　处：《动物营养学报》2019年第10期4667-4674,共8页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家自然科学基金(31860652)

摘　　要：本试验旨在研究不同浓度亚油酸(LA)对二维(2D)和三维(3D)培养模式下奶牛乳腺上皮细胞(BMECs)形态及乳蛋白合成相关基因表达的影响。试验采用单因子完全随机试验设计,将P3代的BMECs接种在铺有基质胶(Matrigel)的6孔板中,培养至第3天时,然后在培养基中添加不同浓度(0、40和80μmol/L)的LA作用48 h。在2D和3D培养模型的BMECs中添加相同浓度的LA,比较2D和3D培养模型中BMECs的细胞形态,并采用实时荧光定量法测定乳蛋白合成相关基因的表达。结果表明:1)BMECs在2D培养模型中呈现鹅卵石的形态,而在3D培养模型中呈现腺泡样结构。2)BMECs中乳蛋白合成相关基因在3D培养模型中的表达量显著高于其在2D培养模型中的表达量(P<0.05)。3)α-酪蛋白(CSN1S1)、信号转导转录激活因子5(STAT5A)、真核翻译启始因子4E结合蛋白1(EIF4EBP1)和核糖体p70s6激酶(RPS6K1)的基因表达量在2D和3D培养模型中的变化趋势一致,而κ-酪蛋白(CSN3)、哺乳动物雷帕霉素靶蛋白(mTOR)和丝氨酸/苏氨酸蛋白激酶(AKT1)的基因表达量在2D和3D培养模型中却呈现相反的趋势。综上,对于CSN3、mTOR和AKT1的基因表达量的研究,3D培养模型为更适宜的模型。This study aims to evaluate the effects different concentrations of linoleic acid(LA)on expression of milk protein synthesis related gene in two-dimension(2D)and three-dimension(3D)cultured bovine mammary epithelial cells(BMECs).A single factorial experimental design was used in the experiment,the P3 generation BMECs were seeded in 6-well plates with Matrigel spread,and cultured for 3 d,next,different concentrations(0,40 and 80μmol/L)of LA were supplemented in the culture medium for 48 h.The same concentration of LA was supplemented in BMECs in 2D and 3D cultured models,to compare the cell morphology in BMECs in 2D and 3D cultured models,and the expression of milk protein synthesis related genes were determined by real-time fluorescence quantitative method.The results showed as follow:1)BMECs showed cobblestone morphology in 2D culture model and alveoli-like structures in 3D culture model.2)The expression of milk protein synthesis related genes in BMECs in 2D culture model was significantly higher than that in 3D culture model(P<0.05).3)The expression ofα-casein(CSN1S1),signal transduction activator 5(STAT5A),eukaryotic translation initiation factor 4E binding protein 1(EIF4EBP1)and ribosomal p70s6 kinase(RPS6K1)in 2D and 3D culture models were the same,however,the expression ofκ-casein(CSN3),mammalian target of rapamycin(mTOR)and serine/threonine kinase(AKT1)in 2D and 3D culture models were showed an opposite trend.To sum up,3D culture model is a more appropriate model for the study related to CSN3,mTOR and AKT1 genes expression.

关 键 词：奶牛 三维细胞培养模型 亚油酸 乳蛋白合成 

分 类 号：S823[农业科学—畜牧学]